Integrated serum pharmacochemistry and serum pharmacology to investigate the active components and mechanism of Bushen Huoxue Prescription in the treatment of diabetic retinopathy.

This study aimed to investigate the active components of Bushen Huoxue Prescriptions (BHP), and further clarify the mechanism by the integration of serum pharmacochemistry and serum pharmacology based on spectrum-effect relationship in vivo. In this paper, the components absorbed into serum were analyzed by ultra-high performance liquid chromatography-quadrupole-Exactive Orbitrap-high resolution mass spectrometry (UPLC-Q-Exactive Orbitrap-HRMS). And Müller cells were chosen as target cells to further investigate the mechanism. After cell purification, the well-grown cells were identified by Hematoxylin-eosin staining (HE) staining and immunofluorescence assay, such as glial fibrillary acidic protein (GFAP) and glutamine synthetase (GS). The logarithmic phase cells were divided into normal group, model group and 12 BHP groups. The hyperglycemic and hypoxic model was induced by 50 mmol/L glucose and 1 mmol/L sodium disulfite. Enzyme-linked immunesorbnent assay (ELISA) was used to detect the expressions of five factors closely related to DR, named vascular endothelial growth factor (VEGF), hypoxia-inducible factor1-alpha (HIF-1α), protein kinase C-ß (PKC-ß), angiopoietin-2 (ANG-2) and transforming growth factor-ß (TGF-ß). Finally, the spectrum-effect relationship was investigated to screen the active components of BHP by partial least squares regression (PLSR). The results showed that 83 metabolic components, containing 30 prototypes and 53 metabolites were found in BHP serum. 12 characteristic common peaks were chosen to establish spectrum-effect relationship. Significantly, all the 12 BHP serum exhibited stronger inhibition on the expression of VEGF, PKC-ß, and ANG-2, and the expression of VEGF, PKC-ß, ANG-2 was chosen to establish the spectrum-effect relationship in vivo. The results of PLSR revealed that the content of methylation and sulfuration of caffeic acid, dehydroxylation and sulfation of Danshensu, daidzein, O-demethylangolanolin, cryptotanshinone, tanshinone IIA and protopanaxatriol were inversely correlated with VEGF expression of Müller cells; the areas of dihydrocaffeic acid, methylation and sulfuration of caffeic acid, dehydroxylation and sulfation of Danshensu, daidzein, cryptotanshinone, tanshinone IIA were negative correlation with the expression of PKC-ß; while the coefficient of hydroxytyrosol sulfation, R-equol, O-demethylangolanolin, dihydrotanshinone IIA, hydrated cryptotanshinone, protopanaxatriol showed negative correlation with the expression of ANG-2. The above results indicated that cryptotanshinone, tanshinone IIA, daidzein and protopanaxatriol need be focused in our future research. In addition, this research idea provides feasible ways to investigate and determine pharmacodyamic material basis and screen the Q-markers of TCM and its formulas.

Exploring the quality markers and mechanism of Bushen Huoxue Prescription in prevention and treatment of diabetic retinopathy based on Chinmedomics strategy.

ETHNOPHARMACOLOGICAL RELEVANCE: Chinese herbal medicines have complex chemical composition; therefore, revealing the effective substances of Chinese herbal medicine becomes a prerequisite for scientific elucidation of the mechanism of action of Bushen Huoxue Prescription (BHP) against diabetic retinopathy (DR) and the development of new drugs. AIM OF THE STUDY: The Chinmedomics technique was used to evaluate the pharmacodynamic ingredients and mechanism of action of BHP against DR rats. MATERIALS AND METHODS: The overall physiological condition of the rats, including body weight, blood glucose, inflammatory factor levels, histological staining, and urine metabolic profile were examined to evaluate the model and its effects. The chemical composition of BHP in vivo and ex vivo was fully analyzed utilizing UPLC-Q-Exactive Orbitrap MS in conjunction with TCM serum pharmacochemistry. Finally, correlation analysis between biomarkers, and serum migration components was used to identify Quality markers (Q-markers) that were significantly associated with effectiveness. RESULTS: The UPLC-Q-Exactive Orbitrap MS platform was used to identify a total of 29 chemicals in serum, 17 of which were highly linked with effectiveness and can be potentially employed as pharmacodynamic substances for BHP against DR. In addition, 14 biomarkers related to galactose metabolism, starch and sucrose metabolism, pantothenate and CoA biosynthesis, glycine, serine, and threonine metabolism were identified. These pathways reveal that DR may be inextricably linked to levels of oxidative stress and inflammation in the organism. Finally, five active ingredients were identified as potential Q-markers of BHP against DR, namely ajugol, protocatechuic acid, tanshinone IIA, panaxatriol and puerarin. CONCLUSION: This study successfully clarified the efficacy and Q-markers of BHP through the Chinmedomics strategy, which is of great significance in determining the quality standards of BHP.

Accelerated oxidation of 2,4,6-trichlorophenol in Cu(II)/H2O2/Cl- system: A unique "halotolerant" Fenton-like process?

The roles of chloride in enhanced oxidative degradation of refractory organic pollutants are recently identified in the Cu(II)/H2O2/Cl- system, but the identity of the reactive oxidants and potential conversion of inorganic chloride to organochlorine in such oxidizing environment remain obscure. Here we report that Cu(II)/H2O2/Cl- system is a unique "halotolerant" Fenton-like process that works most efficiently in saline water among the five tested redox-active metals ions (i.e. Cr(VI), Ce(III), Co(II), Mn(II) and Cu(II)). The observed pseudo first-order rate constant for 2,4,6-trichlorophenol (TCP) degradation was linearly correlated with the elevated Cl- content. The TCP degradation rate at [Cl-]0 = 1000 mM by the Cu(II)/H2O2 system was approximately 46-fold higher than that at [Cl-]0 = 5 mM. The optimal mineralization rate of TCP and percentage of absorbable organic halogens (AOX) decrease were 31.6% and 63.8%, respectively, in the tested Cu(II)/H2O2/Cl- system. However, the detection of fused chlorinated byproducts (i.e. chloro-anthracene-pentaol, dioxine, chlorinated dibenzofuran) reminds us of cautiousness in evaluating the applicability of Cu(II)-catalyzed Fenton-like reaction, particularly while it is to be applied to the treatment of wastewater contaminated with chlorophenols. Two independent models (i.e. "Cu(III) model" and "OH model") were developed to describe the kinetics of Cu(II)/H2O2/Cl- system. The failure of "OH model" to rationalize the observed AOX decay has disproved the "OH model" through reductio ad absurdum. The ability of "Cu(III) model" to adequately explain the experimental data demonstrates that Cu(III)-chloro complexes, rather than OH, is the major product resulting from reactions between Cu(I)-chloro complexes and H2O2 at neutral pH.

An often-overestimated adverse effect of halides in heat/persulfate-based degradation of wastewater contaminants.

Halides (X-) in the industrial wastewater are usually thought to adversely affect the degradation kinetics and mineralization rates in several SO4--based advanced oxidation processes. However, their unfavorable effects might be overestimated, particularly the heat/persulfate (PS) system as tested in the present study. Here the degradation of phenol, benzoic acid, coumarin and acid orange 7 (AO7) was examined with the presence of chloride or bromide in a heat/PS process. Cl- was found to have a dual effect (inhibition followed by enhancement) on the decomposition rates of organic pollutants, whereas the effects of Br- are insignificant within the tested concentration (0-0.2 mM). However, some chlorinated or brominated compounds were still identified in this heat/PS system. Unexpectedly, the mineralization rates of AO7, phenol, benzoic acid and coumarin were not apparently inhibited. In addition, the formation of adsorbable organic halogen (AOX) in the heat/PS system was much less than those in the peroxymonosulfate (PMS)/Cl- or PMS/Br- systems. According to the results of kinetic modeling, SO4- was the dominating radical for AO7 degradation without Cl- or Br-, but Cl2- was the main oxidant in the presence of Cl-, SO4-, Br and Br2- were responsible for the oxidation of AO7 in the presence of Br-. The present study assumes that X2/HOX, rather than halogen radicals, is responsible for the enhanced formation of organohalogens. These findings are meaningful to evaluate the PS-based technologies for the high-salinity wastewater and to develop useful strategies for mitigating the negative effects of halides in advanced oxidation processes (AOPs).

Chlorine incorporation into dye degradation by-product (coumarin) in UV/peroxymonosulfate process: A negative case of end-of-pipe treatment.

Recently, UV/peroxymonosulfate (PMS) seems as a panacea for the treatment of recalcitrant organic pollutants; however, the presence of high concentration of chloride in saline wastewater indeed complicates this end-of-pipe technology. Here a negative case of UV/PMS for the treatment of one of secondary degradation byproducts of dyes (coumarin, COU) is demonstrated. The removal rate of COU is reduced by addition of Cl- (0-10 mM). Further increase in Cl- content favors a rapid COU degradation, whereas Cl- involvement seems to open a "Pandora's box": 1) a variety of chlorinated organic intermediates such as 4-chloroisocoumarin and 5-chloro-2-hydroxy-benzaldehyde are identified; 2) Accumulation and relative increase of absorbable organic halogen (AOX) with reaction time in the presence of high levels of chloride are observed; 3) the acute toxicity of the treated COU solution increases; 4) mineralization rate of COU decreases with the increasing [Cl-]. The fluorescence intensity in the UV/PMS/COU system declines with the addition of Cl-, implying the scavenging effects of chloride on hydroxyl radicals. The possible reaction pathways of COU are discussed. These findings highlight the imperativeness of minimizing auxiliary salt dosages in dyeing processes (i.e., source reduction) and developing new end-of-pipe technologies that can work in a saline environment.

Removal of EMG Artifacts from Multichannel EEG Signals Using Combined Singular Spectrum Analysis and Canonical Correlation Analysis.

Electroencephalography (EEG) signals collected from human scalps are often polluted by diverse artifacts, for instance electromyogram (EMG), electrooculogram (EOG), and electrocardiogram (ECG) artifacts. Muscle artifacts are particularly difficult to eliminate among all kinds of artifacts due to their complexity. At present, several researchers have proved the superiority of combining single-channel decomposition algorithms with blind source separation (BSS) to make multichannel EEG recordings free from EMG contamination. In our study, we come up with a novel and valid method to accomplish muscle artifact removal from EEG by using the combination of singular spectrum analysis (SSA) and canonical correlation analysis (CCA), which is named as SSA-CCA. Unlike the traditional single-channel decomposition methods, for example, ensemble empirical mode decomposition (EEMD), SSA algorithm is a technique based on principles of multivariate statistics. Our proposed approach can take advantage of SSA as well as cross-channel information. The performance of SSA-CCA is evaluated on semisimulated and real data. The results demonstrate that this method outperforms the state-of-the-art technique, EEMD-CCA, and the classic technique, CCA, under multichannel circumstances.

Deciphering the degradation/chlorination mechanisms of maleic acid in the Fe(II)/peroxymonosulfate process: An often overlooked effect of chloride.

In recent years, a significant effort has been devoted into investigating the effects of chloride on the degradation kinetics of aromatic pollutants. The impact of chloride on the decomposition of short-chain carboxylic acid intermediates from aromatics degradation has often been overlooked. In this study the roles of chloride in the oxidation of maleic acid (MA) in the Fe(II)/peroxymonosulfate (PMS) process was investigated. Degradation efficiency, reaction intermediates, adsorbable organic halogen (AOX) accumulation and mineralization were examined. The chloride ion (Cl-) was found to have an overall negative impact on MA degradation and mineralization in the Fe(II)/PMS system. The presence of Cl- led to the formation of chlorinated by-products and a high production of AOX. The mineralization of MA was decreased with increasing Cl- concentrations. Kinetic modeling demonstrated the impact of various radicals largely depended on the concentration of Cl-. The significance of Cl2•- or Cl2 for MA destruction was enhanced with increasing Cl- content, and overwhelmed that of SO4•- when the Cl- concentration was over 5 mM. In the absence of Cl-, SO4•- was the primary radical responsible for MA oxidation. A possible degradation pathway is proposed (cis-trans isomerization, decarboxylation and halogenations processes). These results may help to understand the full oxidation pathways of refractory aromatic compounds and the mechanism of chlorinated by-products formation in industrial saline wastewater treatment.

Effects of chloride on PMS-based pollutant degradation: A substantial discrepancy between dyes and their common decomposition intermediate (phthalic acid).

A considerable effort has been devoted to elucidating the roles of chloride in oxidative degradation and chlorination of dyes. However, few investigations are available on kinetic analysis and transformation pathways of secondary degradation byproducts of dyes in saline wastewater treatment. Here the impact of chlorine on the degradation rate of phthalic acid, a typical dye degradation intermediate, by the Co2+/peroxymonosulfate (PMS) process was examined. Degradation efficiency, intermediate products, AOX (adsorbable organic halogen) formation and mineralization were considered. An overall negative impact was observed within the concentration of Cl- up to 100 mM, differing from the dual effect of chloride on dye degradation process as previously observed. The presence of high levels of Cl- led to a low production of AOX and a reduction of the formation of chlorinated by-products. The mineralization was also restrained when the Cl- concentration was increased. Degradation pathways for these processes are proposed. These findings provide valuable information about the degradation pathways of dyes and about the formation mechanism of chlorinated by-products in industrial saline wastewater treatment.