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A simple and accurate analytical method was developed for simultaneous quantification of three steroidal saponins in the roots of Ophiopogon japonicus via high-performance liquid chromatography (HPLC) with mass spectrometry (MS) in this study. Separation was performed on a Tigerkin C18 column and detection was performed by mass spectrometry. A mobile phase consisted of 0.02% formic acid in water (v/v) and 0.02% formic acid in acetonitrile (v/v) was used with a flow rate of 0.5 mL min−1. The quantitative HPLC–MS method was validated for linearity, precision, repeatability, stability, recovery, limits of detection and quantification. This developed method provides good linearity (r >0.9993), intra- and inter-day precisions (RSD <4.18%), repeatability (RSD <5.05%), stability (RSD <2.08%) and recovery (93.82–102.84%) for three steroidal saponins. It could be considered as a suitable quality control method for O. japonicus. [ABSTRACT FROM AUTHOR]

A new steroidal glycoside (1), (25R)-14α, 17α-hydroxyspirost-5-en-3β-yl 3-O-α-L-rhamnpyranosyl-(1 → 2)-β-D-glucopyranosyl-(1 → 3)-β-D-glucopyranoside, together with three known steroidal glycosides, (25R)-3β-hydroxyspirost-5-en-1β-yl-3-O-α-L-rhamnopyranosyl-(1 → 2)-O-β-D-xylopyranosyl-(1 → 3)-α-L-arabinopyranoside (2), Cixi-ophiopogon B (3) and Cixi-ophiopogon A (4), were obtained from the tuberous roots of Ophiopogon japonicus (Liliaceae). Compound 2 was isolated from the Ophiopogon genus for the first time. Their structures were identified on the basis of extensive mass and nuclear magnetic resonance spectroscopic analysis. [ABSTRACT FROM AUTHOR]

Background: Proliferative vitreoretinopathy (PVR) is a severe blinding complication of retinal detachment surgery. Increasing evidence demonstrate that PVR is associated with oxidative stress. Scutellarin is a natural flavone compound that has been reported to have anti-oxidative activity. However, the effect of scutellarin on PVR remains unknown. In the current study, we assessed the effect of scutellarin on hydrogen peroxide (H2O2)-induced oxidative injury in human retinal pigment epithelium cells (ARPE-19). Methods: ARPE-19 cells were pretreated with different concentrations of scutellarin for 2 h, and then challenged with H2O2 (1 mM) for 24 h. The levels of reactive oxygen species (ROS), malondialdehyde (MDA) and superoxide dismutase (SOD) and glutathione (GSH) activity were measured to assess the level of oxidative stress. Flow cytometry was performed to detect the apoptosis rate of ARPE-19 cells. Expression levels of bcl-2, bax, cleaved-caspase-3, p-JAK2, JAK2, p-STAT3, and STAT3 were measured using western blot. Results: Our results revealed that scutellarin improved the cell viability of H2O2-induced ARPE-19 cells. Scutellarin alleviated the H2O2-induced oxidative stress in ARPE-19 cells, which was illustrated by reduced levels of ROS and MDA, accompanied by increased SOD activity and GSH level. The increased apoptosis rate of ARPE-19 cells caused by H2O2 induction was significantly decreased after scutellarin treatment. H2O2 treatment resulted in significant increase in bax expression and decrease in bcl-2 expression, while the changes in the expressions of bax and bcl-2 were reversed by scutellarin treatment. In addition, scutellarin promoted the activation of JAK2/STAT3 signaling pathway in H2O2-induced ARPE-19 cells. Suppression of JAK2/STAT3 signaling pathway abolished the protective effects of scutellarin on H2O2-induced ARPE-19 cells. Conclusion: These findings suggested that scutellarin was capable for alleviating H2O2-induced oxidative damage in ARPE-19 cells, which might be ascribed to the activation of JAK2/STAT3 signaling pathway. [ABSTRACT FROM AUTHOR]

Reactive oxygenation species (ROS) generated from reperfusion results in cardiac injury through apoptosis and inflammation, while PKR has the ability to promote apoptosis and inflammation. The aim of the study was to investigate whether PKR is involved in hydrogen peroxide (H2O2) induced neonatal cardiac myocytes (NCM) injury. In our study, NCM, when exposed to H2O2, resulted in persistent activation of PKR due to NCM endogenous RNA. Inhibition of PKR by 2-aminopurine (2-AP) or siRNA protected against H2O2 induced apoptosis and injury. To elucidate the mechanism, we revealed that inhibition of PKR alleviated H2O2 induced apoptosis companied by decreased caspase3/7 activity, BAX and caspase-3 expression. We also revealed that inhibition of PKR suppressed H2O2 induced NFκB pathway and NLRP3 activation. Finally, we found ADAR1 mRNA and protein expression were both induced after H2O2 treatment through STAT-2 dependent pathway. By gain and loss of ADAR1 expression, we confirmed ADAR1 modulated PKR activity. Therefore, we concluded inhibition of PKR protected against H2O2-induced injury by attenuating apoptosis and inflammation. A self-preservation mechanism existed in NCM that ADAR1 expression is induced by H2O2 to limit PKR activation simultaneously. These findings identify a novel role for PKR/ADAR1 in myocardial reperfusion injury. [ABSTRACT FROM AUTHOR]

Aims: Plasma S-adenosylhomocysteine (SAH) level is positively associated with cardiovascular risk. However, the relationship between plasma SAH levels and the risk of all-cause and cardiovascular mortality remains unknown. This study aimed to explore the relationship between plasma SAH levels and the risk of all-cause and cardiovascular mortality in patients with coronary artery disease (CAD)., Methods: Plasma SAH levels were measured in 1553 patients with CAD. The association between plasma SAH level and the risk of all-cause and cardiovascular mortality was estimated using Cox Proportional hazards regression models., Results: Relative to participants in the lowest quartile of plasma SAH levels, those in the highest quartile of plasma SAH levels had a higher risk of all-cause death (adjusted Hazard Ratio [HR], 2.15; 95% CI, 1.54-3.01; P＜0.001) and cardiovascular death (adjusted HR, 2.20; 95% CI, 1.49-3.25; P=0.001) in the age- and sex-adjusted model. The results of the multivariable adjusted analysis were similar (all-cause death [adjusted HR, 1.81; 95% CI, 1.27-2.58; P=0.002] and cardiovascular death [adjusted HR, 1.84; 95% CI, 1.21-2.79; P=0.031]). The age- and sex-adjusted HRs for each 1 SD increase in plasma SAH level were 1.30 (95% CI, 1.22-1.38) for all-cause mortality, and 1.34 (95% CI, 1.25-1.43) for cardiovascular mortality, respectively. A 1 SD increase in the SAH level was associated with a 25% higher risk of total death (adjusted HR, 1.25; 95% CI, 1.17-1.34) and a 29% greater risk of cardiovascular death (adjusted HR, 1.29; 95% CI, 1.20-1.39) in multivariable adjusted analysis., Conclusions: We found that the plasma SAH level is positively correlated with the risk of all-cause and cardiovascular mortality in patients with CAD in both age- and sex-adjusted and multivariable-adjusted models.

Increasing radical yields to reduce UV fluence requirement for achieving targeted removal of micropollutants in water would make UV-based advanced oxidation processes (AOPs) less energy demanding in the context of United Nations' Sustainable Development Goals and carbon neutrality. We herein demonstrate that, by switching the UV radiation source from conventional low-pressure UV at 254 nm (UV 254 ) to emerging Far-UVC at 222 nm (UV 222 ), the fluence-based concentration of HO • in the UV/peroxydisulfate (UV/PDS) AOP increases by 6.40, 2.89, and 6.00 times in deionized water, tap water, and surface water, respectively, with increases in the fluence-based concentration of SO 4 •- also by 5.06, 5.81, and 55.47 times, respectively. The enhancement to radical generation is confirmed using a kinetic model. The pseudo-first-order degradation rate constants of 16 micropollutants by the UV 222 /PDS AOP in surface water are predicted to be 1.94-13.71 times higher than those by the UV 254 /PDS AOP. Among the tested water matrix components, chloride and nitrate decrease SO 4 •- but increase HO • concentration in the UV 222 /PDS AOP. Compared to the UV 254 /PDS AOP, the UV 222 /PDS AOP decreases the formation potentials of carbonaceous disinfection byproducts (DBPs) but increases the formation potentials of nitrogenous DBPs.

Effective and affordable disinfection technology is one key to achieving Sustainable Development Goal 6. In this work, we develop a process by integrating Far-UVC irradiation at 222 nm with free chlorine (UV 222 /chlorine) for rapid inactivation of the chlorine-resistant and opportunistic Aspergillus niger spores in drinking water. The UV 222 /chlorine process achieves a 5.0-log inactivation of the A. niger spores at a chlorine dosage of 3.0 mg L -1 and a UV fluence of 30 mJ cm -2 in deionized water, tap water, and surface water. The inactivation rate constant of the spores by the UV 222 /chlorine process is 0.55 min -1 , which is 4.6-fold, 5.5-fold, and 1.8-fold, respectively, higher than those of the UV 222 alone, chlorination alone, and the conventional UV 254 /chlorine process under comparable conditions. The more efficient inactivation by the UV 222 /chlorine process is mainly attributed to the enhanced generation of reactive chlorine species (e.g., 6.7 × 10 -15 M of Cl • ) instead of hydroxyl radicals from UV 222 photolysis of chlorine, which is verified through both experiments and a kinetic model. We further demonstrate that UV 222 photolysis damages the membrane integrity and benefits the penetration of chlorine and radicals into cells for inactivation. The merits of the UV 222 /chlorine process over the UV 254 /chlorine process also include the more effective inhibition of the photoreactivation of the spores after disinfection and the lower formation of chlorinated disinfection byproducts and toxicity.

Far-UVC radiation is an emerging tool for combating pathogenic microorganisms in water, but its vulnerability to water matrix components remains unclear. We herein report the critical impacts of nitrate during Far-UVC disinfection of water. Nitrate at environmentally relevant concentrations (0.5-10.0 mg-N L -1 ) significantly inhibits Escherichia coli inactivation by Far-UVC radiation at 222 nm, via prolonging the "lag phase" of inactivation and reducing the inactivation rate constants by 1.08-2.74 times, while it shows negligible impact on E. coli inactivation by UVC radiation at 254 nm. The inhibitory impact of nitrate on Far-UVC disinfection is attributed to its strong light-shielding effect. Although hydroxyl radicals and reactive nitrogen species are generated from Far-UVC photolysis of nitrate at high concentrations of 10 -13 and ∼10 -7 M, respectively, those radicals are unable to compensate for the light-shielding effect of nitrate on E. coli inactivation. Moreover, reactive nitrogen species lead to the formation of nitrogenous byproducts, which increase the genotoxicity of the water. The findings advance the fundamental photochemistry and radical chemistry of nitrate at 222 nm and provide useful insights to guide the operation of Far-UVC in treating nitrate-containing water.

Background: With the diabetes mellitus (DM) prevalence increasing annually, the human grading of retinal images to evaluate DR has posed a substantial burden worldwide. SmartEye is a recently developed fundus image processing and analysis system with lesion quantification function for DR screening. It is sensitive to the lesion area and can automatically identify the lesion position and size. We reported the diabetic retinopathy (DR) grading results of SmartEye versus ophthalmologists in analyzing images captured with non-mydriatic fundus cameras in community healthcare centers, as well as DR lesion quantitative analysis results on different disease stages., Methods: This is a cross-sectional study. All the fundus images were collected from the Shanghai Diabetic Eye Study in Diabetics (SDES) program from Apr 2016 to Aug 2017. 19,904 fundus images were acquired from 6013 diabetic patients. The grading results of ophthalmologists and SmartEye are compared. Lesion quantification of several images at different DR stages is also presented., Results: The sensitivity for diagnosing no DR, mild NPDR (non-proliferative diabetic retinopathy), moderate NPDR, severe NPDR, PDR (proliferative diabetic retinopathy) are 86.19, 83.18, 88.64, 89.59, and 85.02%. The specificity are 63.07, 70.96, 64.16, 70.38, and 74.79%, respectively. The AUC are PDR, 0.80 (0.79, 0.81); severe NPDR, 0.80 (0.79, 0.80); moderate NPDR, 0.77 (0.76, 0.77); and mild NPDR, 0.78 (0.77, 0.79). Lesion quantification results showed that the total hemorrhage area, maximum hemorrhage area, total exudation area, and maximum exudation area increase with DR severity., Conclusions: SmartEye has a high diagnostic accuracy in DR screening program using non-mydriatic fundus cameras. SmartEye quantitative analysis may be an innovative and promising method of DR diagnosis and grading.

Oxidative stress induced by hydrogen peroxide (H 2 O 2 ) triggers human lens epithelial cell (HLEC) apoptosis and initiates cataract formation. Oxyresveratrol (Oxy) was reported to possess antioxidant and free radical scavenging activities. Herein, we investigated the effects of Oxy on H 2 O 2 -induced oxidative stress and apoptosis in HLECs and the associated mechanisms. Cell viability was detected by MTT assay. The oxidative damage was assessed by measuring the activities of superoxide dismutases-1 (SOD-1), catalase (CAT), glutathione reductase (GSH), and malondialdehyde (MDA). Apoptosis was analyzed by flow cytometry analysis. The changed expressions of heme oxygenase-1 (HO-1) and protein kinase B (Akt) pathways were evaluated by qRT-PCR and western blot. We found that exposure to H 2 O 2 dose-dependently reduced cell viability, and induced oxidative stress and apoptosis in HLECs, which were reversed by pretreatment with Oxy. Oxy increased p-Akt and HO-1 expressions in H 2 O 2 -stimulated HLECs. Akt and HO-1 expressions form a regulatory axis and Oxy activated the Akt/HO-1 pathway in H 2 O 2 -stimulated HLECs. Inhibition of the Akt/HO-1 pathway by LY294002 or ZnPP attenuated the effects of Oxy on oxidative stress and apoptosis in H 2 O 2 -stimulated HLECs. In conclusion, Oxy protected H 2 O 2 -induced oxidative stress and apoptosis through activating the Akt/HO-1 pathway, suggesting the protective effect of Oxy against H 2 O 2 -induced cataract., (Copyright © 2019 The Authors. Production and hosting by Elsevier B.V. All rights reserved.)

Background: Proliferative vitreoretinopathy (PVR) is a severe blinding complication of retinal detachment surgery. Increasing evidence demonstrate that PVR is associated with oxidative stress. Scutellarin is a natural flavone compound that has been reported to have anti-oxidative activity. However, the effect of scutellarin on PVR remains unknown. In the current study, we assessed the effect of scutellarin on hydrogen peroxide (H 2 O 2 )-induced oxidative injury in human retinal pigment epithelium cells (ARPE-19)., Methods: ARPE-19 cells were pretreated with different concentrations of scutellarin for 2 h, and then challenged with H 2 O 2 (1 mM) for 24 h. The levels of reactive oxygen species (ROS), malondialdehyde (MDA) and superoxide dismutase (SOD) and glutathione (GSH) activity were measured to assess the level of oxidative stress. Flow cytometry was performed to detect the apoptosis rate of ARPE-19 cells. Expression levels of bcl-2, bax, cleaved-caspase-3, p-JAK2, JAK2, p-STAT3, and STAT3 were measured using western blot., Results: Our results revealed that scutellarin improved the cell viability of H 2 O 2 -induced ARPE-19 cells. Scutellarin alleviated the H 2 O 2 -induced oxidative stress in ARPE-19 cells, which was illustrated by reduced levels of ROS and MDA, accompanied by increased SOD activity and GSH level. The increased apoptosis rate of ARPE-19 cells caused by H 2 O 2 induction was significantly decreased after scutellarin treatment. H 2 O 2 treatment resulted in significant increase in bax expression and decrease in bcl-2 expression, while the changes in the expressions of bax and bcl-2 were reversed by scutellarin treatment. In addition, scutellarin promoted the activation of JAK2/STAT3 signaling pathway in H 2 O 2 -induced ARPE-19 cells. Suppression of JAK2/STAT3 signaling pathway abolished the protective effects of scutellarin on H 2 O 2 -induced ARPE-19 cells., Conclusion: These findings suggested that scutellarin was capable for alleviating H 2 O 2 -induced oxidative damage in ARPE-19 cells, which might be ascribed to the activation of JAK2/STAT3 signaling pathway.

In donor hearts from mini pigs, overtime cold preservation and ischemia-reperfusion injury cause poor graft quality and impaired heart function. Blockage of complement, apoptosis, and inflammation is considered a strategy for attenuating ischemia-reperfusion injury and protecting cardiac function. Minipig donor hearts were perfused and preserved in Celsior solution or transfection reagent containing Celsior solution with scramble siRNA or siRNAs targeting complement 3, caspase-8, caspase-3, and nuclear factor κB-p65 genes at 4°C and subsequently hemo-reperfused ex vivo (38°C) or transplanted into recipients. The protective effect of the siRNA solution was evaluated by measuring cell apoptosis, structural alteration, protein markers for tissue damage and oxidative stress, and cardiac function. We found a reduction in cell apoptosis, myocardial damage, and tissue inflammation by reduced biochemistry and markers and protein expression of proinflammatory cytokines and improvement in cardiac function, as shown by the improved hemodynamic indices in 12-hr-preserved siRNA-treated hearts of both ex vivo and orthotopic transplantation models. These findings demonstrate that blockade of inflammation and apoptosis pathways using siRNA can prolong cold preservation time and better protect donor heart function in cardiac transplantation of large animals, which may be beneficial for human heart preservation., (Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.)

Abnormal phenotypic modulation of vascular smooth muscle cells (VSMCs) is a hallmark of cardiovascular diseases such as atherosclerosis, hypertension and restenosis after angioplasty. Transcription factors have emerged as critical regulators for VSMCs function, and recently we verified inhibiting transcription factor Gax was important for controlling VSMCs proliferation and migration. This study aimed to determine its role in phenotypic modulation of VSMCs. Western blot revealed that overexpression of Gax increased expression of VSMCs differentiation marker genes such as calponin and SM-MHC 11. Then, Gax overexpression potently suppressed proliferation and migration of VSMCs with or without platelet-derived growth factor-induced-BB (PDGF-BB) stimuli whereas Gax silencing inhibited these processes. Furthermore, cDNA array analysis indicated that Rap1A gene was the downstream target of Gax in human VSMCs. And overexpression of Gax significantly inhibited expression of Rap1A in VSMCs with or without PDGF-BB stimuli. Moreover, overexpression of Rap1A decreased expression of VSMCs differentiation marker genes and increased proliferation and migration of VSMCs with or without PDGF-BB stimuli. Finally, Gax overexpression significantly inhibited the neointimal formation in carotid artery injury of mouse models, specifically through maintaining VSMCs contractile phenotype by decreasing Rap1A expression. In conclusion, these results indicated that Gax was a regulator of human VSMCs phenotypic modulation by targeting Rap1A gene, which suggested that targeting Gax or its downstream targets in human VSMCs may provide an attractive approach for the prevention and treatment of cardiovascular diseases.

The aim of the present study was to investigate the expression of nesprin-1 protein in MSCs and its effects on the differentiation of rat bone-marrow mesenchymal stem cells (MSCs). Surface-associated antigens of MSCs were detected by flow cytometry. MSC differentiation was induced by treatment with 10 µmol/l 5-azacytidine. Sprague-Dawley rats were anesthetized prior to thoracotomy and subsequent ligation of the left anterior descending coronary artery to establish a model of myocardial infarction. Two weeks following myocardial infarction, DAPI-marked MSCs were injected into the infarcted region in the experimental group, while DMEM was injected into the infarcted region of the control group. Characteristics of the putative cardiac-myogenic cells were evaluated using immunohistochemical and immunofluorescent analysis. The messenger RNA expression levels of cardiac-myogenic specific genes; desmin, α-actinin and cardiac troponin I (cTnI) were detected by reverse transcription quantitative polymerase chain reaction. The expression of nesprin-1 protein in MSCs was identified by immunofluorescence and western blot analysis, prior to and following MSC differentiation. Following differentiation, the MSCs appeared spindle-shaped with irregular processes and were positive for CD90 and CD29, but negative for CD45. Cardiomyocyte-like cells were positive for desmin, α-sarcomeric actin and cTnI. Nesprin protein was detected in the nuclear membrane via immunofluorescence, and following MSC differentiation into cardiomyocyte-like cells, the expression of nesprin protein was significantly higher (*P=0.03<0.05). The results of the present study indicated that MSCs may be differentiated in vitro and in vivo into cells with characteristics commonly attributed to cardiomyocytes. Cardiomyocyte-like cells cultured from bone marrow sources may be potentially useful for repairing the injured myocardium. The results also suggested that, consistent with the results of previous studies, the expression of nesprin-1 protein was higher during the differentiation process of MSCs and may have an important role in mediating MSC differentiation. Elucidation of the role of nesprin-1 in MSC differentiation may aid in the development of novel therapies for the treatment of myocardial ischemia and nesprin-1 genetic deficiencies.

Wnt/β-catenin signalling plays a rate-limiting role in early development of many different organs in a broad spectrum of organisms. In the developing Müllerian duct, Wnt/β-catenin signalling is important for initiation, outgrowth, patterning and differentiation into vagina, cervix, uterus and oviducts. In adult life, sex hormones modulate Wnt/β-catenin signalling in the endometrium to maintain the monthly balance between estrogen-induced proliferation and progesterone-induced differentiation, and enhanced Wnt/β-catenin signalling seems to be involved in endometrial carcinogenesis. However, early in pregnancy enhanced Wnt/β-catenin signalling is prerequisite for proper implantation and invasion of trophoblast cells into endometrium and myometrium thus helping to form a placenta. Overall, it seems that tight control of Wnt/β-catenin signalling in time and space is important for initiation, development and normal function of the female reproductive tract. However, if Wnt/β-catenin signalling is not kept in check, it easily seems to initiate or contribute to development of a number of uterine disorders., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

The authors studied contributors to stress among undergraduate residence hall students at a midwestern, land grant university using a 76-item survey consisting of personal, health, academic, and environmental questions and 1 qualitative question asking what thing stressed them the most. Of 964 students selected at random, 462 (48%) responded to the survey. The authors weighted data to reflect the overall university-wide undergraduate population (55% men, 12% minority or international, and 25% freshmen). Women and US citizens experienced greater stress than did men and non-US citizens, respectively. Frequency of experiencing chronic illness, depression, anxiety disorder, seasonal affective disorder, mononucleosis, and sleep difficulties were significant stress predictors. Although alcohol use was a positive predictor, drug use was a negative predictor of stress. Both a conflict and a satisfactory relationship with a roommate, as well as a conflict with a faculty or staff member, were also significant predictors of stress.