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Aromatic carboxylic acids (ACAs) may be as transformed key metabolites via gut microbiome for playing better pharmacological effects. However, it's rare to achieve high-specificity, high-sensitivity, and high-throughput detection simultaneously, especially, for tracing trace ACAs in gut microbiome. In this work, firstly, a novel dual-template and double-shelled molecularly imprinted 96-well microplates (DDMIPs) was designed and amplified signal for p-hydroxybenzoic acid (PBA) and 3,4,5-trimethoxycinnamic acid (TMA). Additionally, the DDMIPs and a stable isotope labeling derivatization (SILD) method combined with the ultra-high performance liquid chromatography triple quadrupole tandem mass spectrometry (UHPLC-TQ MS) was firstly stepwise integrated, achieving high-effective, high-sensitive, and high-throughput study of gut microbiome metabolism. The whole strategy showed lower limits of detections (LODs) up to 1000 folds than the traditional method, and revealed a more real metabolism-time profile of PBA and TMA by 3-step signal amplification. The platform also laid the foundation for fast, simple, high-selective, high-effective, and high-throughput metabolism and pharmacological research.

To explore the relationship between the severity of hypertension and the imbalanced intestinal flora, inflammatory factors, adiponectin (ADPN) and vascular endothelial function in primary hypertension patients. According to the grading criteria for hypertension, in total of 60 patients with primary hypertension in our hospital from April to July, 2020 were divided into Grade 1 group (n = 20), Grade 2 group (n = 20), and Grade 3 group (n = 20). The feces of the research subjects were collected to extract the deoxyribonucleic acid (DNA) and detect its composition of intestinal flora. Subsequently, the peripheral blood was collected to determine the changes in inflammatory factors interleukin‐2 (IL‐2), IL‐4, tumor necrosis factor‐α (TNF‐α) and IL‐1β, serum immunoglobulin G (IgG) and IgM, ADPN and vascular endothelial function‐related endothelin‐1 (ET‐1), nitric oxide (NO), vascular endothelial growth factor (VEGF), and intercellular adhesion molecule‐1 (ICAM‐1). There were no significant differences in the gender, age, and body mass index (BMI), the proportion of smokers, diet habit, probiotics and antihypertensive medication use, and number of diabetic cases among groups (p > 0.05). We found an inverse association between blood pressure measures and microbial diversity, in particular microbial richness (p < 0.05). Among the four major kinds of intestinal flora, the composition of firmicutes (p < 0.05) and bacteroidetes (p < 0.05) showed obvious differences among the three groups, and they had consistent trends with the changes in the abundance of firmicutes and bacteroidetes. Intestinal flora imbalance is closely related to the severity of hypertension, inflammatory factors, ADPN, and vascular endothelial function.

Organic adhesives (e.g. epoxy resin) have been widely used in the near-surface-mounted (NSM) fibre reinforced polymer (FRP) strengthening system for reinforced concrete (RC) structures. However, the drawbacks of the organic adhesives in terms of high-temperature resistance and compatibility with concrete urge the development of inorganic adhesives (e.g. cement-based and alkali-activated materials). Moreover, a premature debonding of NSM FRPs from concrete substrate adversely affects the performance of strengthened RC structures, as well as the efficiency of NSM FRPs. Various anchorage strategies have been consequently proposed to improve the bond between the NSM FRPs and the concrete. This paper presents an overview on the bond behaviour between inorganic adhesive based NSM FRPs and concrete. Flexural strengthening of RC beams with inorganic adhesive based NSM FRPs, and the anchorage strategies for NSM FRPs are also summarized. Different from epoxy-based adhesives, the bond strength of inorganic adhesive based NSM FRPs decreases as the groove dimension increases. The RC beams strengthened by inorganic adhesive based NSM FRPs tend to fail due to an intermediate crack debonding of FRPs, which is probably caused by the low deformation capacity of inorganic adhesives. Anchorage strategies, such as adding external wrapping FRP fabrics or internal additional ribs, could significantly enhance the strengthening efficiency and the constructability of NSM FRPs. In the end, recommendations for future studies on the use of inorganic adhesive based NSM FRPs for structural strengthening are outlined.

Cancer and its associated conditions have significant impacts on public health at many levels worldwide, and cancer is the leading cause of death among adults. Peroxisome proliferator-activated receptor α (PPARα)-specific agonists, fibrates, have been approved by the Food and Drug Administration for managing hyperlipidemia. PPARα-specific agonists exert anti-cancer effects in many human cancer types, including glioblastoma (GBM). Recently, we have reported that the hypoxic state in GBM stabilizes hypoxia-inducible factor-1 alpha (HIF-1α), thus contributing to tumor escape from immune surveillance by activating the expression of the pH-regulating protein carbonic anhydrase IX (CA9). In this study, we aimed to study the regulatory effects of the PPARα agonist fibrate on the regulation of HIF-1α expression and its downstream target protein in GBM. Our findings showed that fenofibrate is the high potency compound among the various fibrates that inhibit hypoxia-induced HIF-1α and CA9 expression in GBM. Moreover, fenofibrate-inhibited HIF-1α expression is mediated by HO-1 activation in GBM cells through the AMP-activated protein kinase (AMPK) pathway. In addition, fenofibrate-enhanced HO-1 upregulation activates SIRT1 and leads to subsequent accumulation of SIRT1 in the nucleus, which further promotes HIF-1α deacetylation and inhibits CA9 expression. Using a protein synthesis inhibitor, cycloheximide, we also observed that fenofibrate inhibited HIF-1α protein synthesis. In addition, the administration of the proteasome inhibitor MG132 showed that fenofibrate promoted HIF-1α protein degradation in GBM. Hence, our results indicate that fenofibrate is a useful anti-GBM agent that modulates hypoxia-induced HIF-1α expression through multiple cellular pathways.

An accurate global potential energy surface (PES) for the HBr2 system has been constructed using the fundamental invariant neural network fitting method based upon 11 698 ab initio energies at the UCCSD(T)/CBS level of theory, with the spin-orbit coupling of the 2P3/2 orbit of the Br atom properly included. The time-dependent wave packet calculations have been performed to study the H + Br2 → HBr + Br reaction on the new PES. The total reaction probabilities for total angular momentum J = 0 for the ground initial state show no threshold due to the submerged barrier height (-0.351 kcal/mol) of the PES. The total integral cross sections (ICS) for reactant Br2 in ro-vibrational states (v0 = 0, j0 = 0, 10, 20, 30; v0 = 1-5, j0 = 0) were calculated for collision energy of up to 0.5 eV. It is found that the initial rotational excitation has a negligible effect on the ICS, and the initial vibrational excitation depresses the reactivity to some extent. The thermal rate constants for the title reaction in the temperature range of 100-1000 K were calculated from the Boltzmann averaging of the v0 = 0-5 rate constants, which overestimated the experimental results to some extent.

Oxazole is an important pharmacophore and exists in the backbone of many bioactive peptide natural products and peptidomimetics. Efficient methods for the synthesis and direct functionalization of complex oxazole-containing peptides are in high demand. Herein, we report the late-stage site-selective functionalization of oxazole-containing peptides via palladium-catalyzed δ-C(sp2)-H olefination of phenylalanine, tryptophan, and tyrosine residues. This strategy utilizes oxazole motifs as internal directing groups and provides access to oxazole-containing peptide macrocycles with bioactivities.

This study aimed to investigate the role of long noncoding RNA (lncRNA) nuclear-enriched abundant transcript 1 (NEAT1) in the development of ALF. We collected blood samples from patients with acute liver failure (ALF) and established an ALF mouse model induced by D-galactosamine/Lipopolysaccharide (D-GalN/LPS) for in vivo studies. Peripheral blood mononuclear cells (PMBCs) induced with LPS were isolated for in vitro experiments. Survival tests, histological analysis, and biochemical indicator assays were conducted. Luciferase assay was performed to determine the binding affinity between microRNA-139 (miR-139) and p53-upregulated modulator of apoptosis (PUMA). Expression of lncRNA NEAT1, enhancer of zeste homolog 2 (EZH2), and PUMA was upregulated, while the expression of miR-139 was downregulated in clinical samples and D-GalN/LPS induced ALF mouse model. LncRNA NEAT1 promoted the enrichment of H3K27me3 on the promoter region of miR-139 via EZH2, which led to suppression of miR-139. The inhibition of miR-139 resulted in the upregulation of its downstream target PUMA. The NEAT1/miR-139/PUMA pathway upregulated the production of pro-inflammatory cytokines, tumor necrosis factor alpha, interleukin (IL)-6, and IL-1β, thereby mediating the progression of ALF. In conclusion, silencing lncRNA NEAT1 upregulated the expression of miR-139 through EZH2, leading to the downregulation of PUMA, which alleviated the development of ALF.

Multidrug resistance (MDR) is one of the major obstacles for clinical effective chemotherapy. In this study, the effects and possible mechanisms of aloe-emodin (AE) were investigated on reversing the adriamycin (ADR)-induced resistance of MCF-7/ADR cells. AE could significantly reverse the ADR resistance in MCF-7/ADR cells. The combination of AE (20 μM) and ADR had no effect on the P-glycoprotein (P-gp) level, but notably promoted the accumulation of ADR in drug-resistant cells. The efflux function of P-gp required ATP, but AE reduced the intracellular ATP level. AE played a reversal role might through inhibiting the efflux function of P-gp. The research result of energy metabolism pathways indicated that combination of AE and ADR could inhibit glycolysis, tricarboxylic acid (TCA) cycle, glutamine metabolism, and related amino acid synthesis pathways. Moreover, we found AE not only reversed ADR-induced resistant but also induced autophagy as a defense mechanism. In addition, the combination of AE and ADR arrested G2/M cell cycle and induced apoptosis through DNA damage, ROS generation, caspase-3 activation. Our study indicated that AE could be a potential reversal agent to resensitize ADR resistant in tumor chemotherapy and inhibiting autophagy might be an effective strategy to further enhance the reversal activity of AE.

The boronate affinity-based oriented and double-shelled surface molecularly imprinted polymers on 96-well microplates (BDMIPs) were designed and applied to high-specific and high-throughput pharmacokinetic (PK) study of rutin and its metabolites from rat plasma without concentration and redissolution. It integrated the advantages of covalent effects-based boronate affinity, noncovalent effects of ethylene imine polymer (PEI) dendrimer, multiple cavities-based double-shelled layers, and multiparallel wells-based 96-well microplates. Furthermore, ultrahigh-performance liquid chromatography triple quadrupole tandem mass spectrometry (UHPLC-MS/MS) was used to accurately quantify targets. It showed lower limits of detection (LODs) up to 100-fold than the conventional method. And PKs of rutin and trace isoquercetin (IQC) were first reported at the same time. The platform can provide a fast, simple, low-cost, high-selective, high-effective, and high-throughput methodological reference for analysis of large-scale samples in the fields of agriculture and food.

Metal complexes exhibit higher stability in water, making metal removal more challenging. In this study, a low-cost, high-efficiency, and multifunctional nanomanganese oxide-modified biochar (BC-Mn...

VPS33B is reported to be a tumor suppressor in hepatocellular carcinoma, nasopharyngeal carcinoma, colon cancer, and lung adenocarcinoma. Here, we observed that reduced VPS33B protein level was an unfavorable factor that promoted the pathogenesis of non-small cell lung cancer (NSCLC) in clinical specimens. We achieved lentivirus-mediated stable overexpression of VPS33B in NSCLC cells. Increased VPS33B reduced cell cycle transition and cell proliferation of NSCLC cells in vivo and in vitro. Knocking down VPS33B restored cell growth. Mechanism analysis indicated that miR-192-3p was induced by VPS33B and acted as a tumor suppressor of cell growth in NSCLC. Further, c-Myc or p53 was identified as a transcription factor that bound to the miR-192-3p promoter and regulated its expression. miR-192-3p directly targeted cell cycle-promoted factor CCNB1 and suppressed NSCLC cell growth. VPS33B modulated c-Myc/p53/miR-192-3p signaling to target CCNB1 by reducing activation of the Ras/ERK pathway. Our study reveals a novel molecular basis for VPS33B as a tumor suppressor to participate in the pathogenesis of NSCLC., Graphical Abstract, The dysfunction of a large number of genes participates in the pathogenesis of NSCLC. Liu and colleagues demonstrated that VPS33B acts as a tumor suppressor to halt NSCLC pathogenesis and elucidated the underlying mechanism by which VPS33B regulates c-Myc/p53/miR-192-3p to target CCNB1 via inactivating the Ras/ERK signal.

In this study, catalytic activity and deactivation mechanism of a carbon-based solid-acid catalyst in the esterification of oleic acid and methanol were investigated. The experimental results showed that the S150-4 catalyst, prepared by sulphonating at 150 °C for 4 h, yielded the highest sulfonic acid density. The conversion yield reached 97.98% under the optimum esterification conditions (i.e. catalyst loading, 10 wt%; methanol/oleic acid molar ratio, 8:1; reaction temperature, 65 °C; reaction time, 8 h) and decreased to 79.19% after four cycles. In the first batch of esterification, the formation of sulfonic esters was the main reason for the deactivation of the catalyst, whereas the leaching of sulphur was the main contributor in the second to fourth batches. Therefore, it is believed that reducing the leaching of sulphur and the formation of sulfonic esters by changing the addition method of methanol is an effective method to extend the service life of the catalyst.

The determination of trace fluorine and chlorine in iron ore commonly requires complicated and time-consuming pretreatment involving alkali fusion, combustion in bombs, and pyrohydrolysis that prev...

Quick identification of the complex composition of traditional Chinese medicine only through liquid-mass spectrometry technology is difficult. Especially the identification of isomers and co-eluting compounds is even more difficult. In this study, an approach of multidimensional data modes based on ultra-performance liquid chromatography coupled with traveling wave ion mobility quadrupole time-of-flight mass spectrometry was proposed to quickly and comprehensively identify the compounds in Platycodi Radix. First, data-independent acquisition, high-definition acquisition, and tandem mass spectrometry acquisition modes were used to acquire integrated multidimensional mass spectral data. Second, summarize the diagnostic ions of compounds according to the fragmentation pathway of references. Third, unknown compounds and isomers were identified via the UNIFI™ software with an in-house library. Finally, a total of 87 compounds were identified, seven compounds were explicitly identified by comparing the retention time and fragment ions with the references. Fourteen compounds were first detected in the Platycodi Radix, four of them tentatively were identified by comparing with previous literature, eight compounds were observed and reported for the first time by comparing typical fragmentation pathway with the known standard substances in this paper. This research strategy has a certain potential for the analysis of complex components of other traditional Chinese medicine.

Hepatocellular carcinoma (HCC) is the most common form of hepatic malignancies. The diagnosis of HCC remains challenging due to the low sensitivity and specificity of the diagnostic method. Exosomes, which are abundant in various proteins from parent cells, play pivotal roles in intercellular communication and have been confirmed as promising sources of disease biomarkers. Herein, we performed a simple but robust proteomic profiling on exosomes derived from 1 μL of serum using a data-independent acquisition (DIA) method for the first time, to screen potential biomarkers for the diagnosis of HCC. Ten pivotal differentially expressed proteins (DEPs) (von Willebrand factor (VWF), LGALS3BP, TGFB1, SERPINC1, HPX, HP, HBA1, FGA, FGG, and FGB) were screened as a potential candidate biomarker panel, which could completely discriminate patients with HCC from normal control (NC). Interestingly, Gene Expression Profiling Interactive Analysis (GEPIA) revealed that the expression levels of four genes increased and those of six genes decreased in HCC tissues compared with normal tissues, which were in concordance with protein expression levels. In conclusion, we screened 10 exosomal proteins holding promise for acting as a potential candidate biomarker panel for detection of HCC through a simple but robust proteomic profiling.

Quercetin is a widely-occurring flavonoid that protects against cancer, and improves memory and cardiovascular functions. However, whether quercetin exhibits therapeutic effects in diabetic retinopathy remains unclear. In this study, we established a rat model of streptozocin-induced diabetic retinopathy. Seventy-two hours later, the rats were intraperitoneally administered 150 mg/kg quercetin for 16 successive weeks. Quercetin markedly increased the thickness of the retinal cell layer, increased the number of ganglion cells, and decreased the overexpression of the pro-inflammatory factors interleukin-1β, interleukin-18, interleukin-6 and tumor necrosis factor-α in the retinal tissue as well as the overexpression of high mobility group box-1 and the overactivation of the NLRP3 inflammasome. Furthermore, quercetin inhibited the overexpression of TLR4 and NF-κBp65, reduced the expression of the pro-angiogenic vascular endothelial growth factor and soluble intercellular adhesion molecule-1, and upregulated the neurotrophins brain-derived neurotrophic factor and nerve growth factor. Intraperitoneal injection of the heme oxygenase-1 inhibitor zinc protoporphyrin blocked the protective effect of quercetin. These findings suggest that quercetin exerts therapeutic effects in diabetic retinopathy possibly by inducing heme oxygenase-1 expression. This study was approved by the Animal Ethics Committee of China Medical University, China (approval No. 2016PS229K) on April 8, 2016.

Schisandra chinensis (Turcz.) Baill Fructus (SCF) is the ripe fruit of Schisandra chinensis (Turcz.) Baill, and is often used as a neuroprotective drink. Modern pharmacological studies have shown that lignans are the main bioactive components responsible for neuroprotection and have potential in the treatment of Alzheimer's disease (AD). However, the mechanism of action of SCF in the treatment of AD from the pharmacokinetics-pharmacodynamics (PK-PD) perspective remains not well established. The purpose of this study is to investigate and compare the pharmacokinetic differences of lignans in normal and AD rats, as well as to investigate their effects on neurotransmitters and their role in the treatment of AD. To achieve this goal, an integrated strategy using LC-MS/MS combined with in vivo microdialysis for the simultaneous determination of lignans of SCF and endogenous neurotransmitters has been developed and validated. The results show that the pharmacokinetic behaviors of ten lignans in the AD group were significantly different from those in the normal group. The AD group had better absorption and slower elimination than the normal group. In addition, the pharmacodynamic results of the Morris water maze (MWM) test, biochemical tests, histopathological examination, as well as immunohistochemistry analysis showed that lignans could improve the learning and memory of AD rats. The oral administration of SCF could restore the levels of the neurotransmitter parameters; seven neurotransmitters showed clockwise or counterclockwise changes with the four lignans in the hippocampal region. Taken together, the PK and PD studies based on in vivo microdialysis sampling might offer novel insights into the mechanisms of action of SCF against AD.

Eleutherococcus senticosus (Rupr. & Maxim.) Maxim. leaves (ESL) were reported to have neuroprotective function and are also used to treat cranial and cerebral traumas as a traditional Chinese medicine and food herbage plant. However, there has been no previous study on ESL treatment for stroke at the level of lipid disorders. To clarify the mechanism of ESL in treating ischemic stroke, this study was carried out from 3 aspects, namely, the regulation of lipid disorders, protection of the nervous system, as well as anti-inflammatory and antioxidant actions. This study established a lipidomics research strategy that was developed by UPLC-Q-TOF/MS analysis. The quantification of neurotransmitters in the serum and brain tissue of rats was performed using UPLC-TQ/MS. Also, we quantified the oxidative stress and inflammatory reaction by measuring the contents of SOD, MDA, TNF-α, IL-6, and IL-10 via the ELISA kits for serum and brain tissue. According to UPLC-Q-TOF/MS-based lipidomics analysis, 27 lipidomics biomarkers were identified in this study, including PC, PE, SM, and TG, which were distributed in various lipid metabolic pathways, including glycerophospholipid, linoleic acid, alpha-linolenic acid, glycerolipid, sphingolipid, and arachidonic acid metabolism pathways. By reversing the changes in the lipid content caused by the disease, ESL has a therapeutic effect on ischemic stroke. Furthermore, quantitative results of neurotransmitters indicated that they can be regulated by ESL. Finally, the results of ELISA showed that ESL can treat ischemic stroke to a certain extent by reducing the oxidative and inflammatory damage. Therefore, ESL may play a therapeutic role in the treatment of ischemic stroke in different ways. This research preliminarily revealed the mechanism of ESL in the treatment of ischemic stroke and provided support for the further application of ESL.

Scutellaria baicalensis Georgi (S. baicalensis) is a traditional Chinese medicine that has been used in the treatment of diabetes for centuries. Although it has curative effects on diabetic nephropathy (DN), the critical role and mechanisms of S. baicalensis in the prevention and management of DN have not been fully elucidated. Thus, in the current study, metabolomics was integrated with a network pharmacology approach to explore the mechanisms of S. baicalensis against DN. To illustrate the metabolic alterations regulated by S. baicalensis during DN progression, an untargeted serum metabolomics approach using ultra-high- performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) was developed. It revealed that 14 serum metabolic biomarkers identified in DN rats were partially reversed by S. baicalensis treatment, which was primarily involved in glycerophospholipid metabolism, arachidonic acid metabolism and retinol metabolism pathways, with 536 extended metabolite-target genes (M-T) interactions observed. Based on network pharmacology analysis, a total of 31 active compounds and 123 compound-target genes (C-T) interactions of S. baicalensis were identified. By integrating M-T with C-T co-network analysis, 18 shared genes were identified as hub targets against DN, including MAPK14, MPO, MMP9, FN1, NOS2 and CASP3, which primarily contributed to biological processes related to inflammatory responses. The molecular docking analysis further verified the interactions between the major active compounds and six predicted proteins encoded by the hub targets, indicating that S. baicalensis may function by targeting inflammatory responses process in DN. Overall, the current work yielded new insights into the molecular mechanisms of active compounds in the action of S. baicalensis against DN progression and suggested a strategy for clarifying the treatment mechanisms of traditional herbal medicines.

Qishen granules, derived from clinical experience formula, has been widely used to improve and treat myocardial ischemic chronic heart failure in China. However, the mechanism of action of Qishen granules in the treatment of chronic heart failure is unclear. This study aimed to discover potential biomarkers of isoproterenol-induced chronic heart failure rats and investigate the potential mechanism of Qishen granules treatment of chronic heart failure. The fecal metabolomics method based on ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry was used to analyze the therapeutic effect and metabolic changes of Qishen granules on chronic heart failure rats. Totally, 17 potential biomarkers were identified, involving bile acid metabolism, fatty acid metabolism, inflammatory response, and amino acid metabolism. For bile acid metabolism, we selected 12 bile acids (two of which were potential biomarkers in nontargeted metabolomics) for quantitative analysis. The quantitative results of bile acids showed that after Qishen granules treatment, the contents of bile acids such as ursodeoxycholic acid and glycodeoxycholic acid were similar to those of health group. This study helps to understand the pathogenesis of isoproterenol-induced chronic heart failure and the therapeutic mechanism of Qishen granules from the perspective of metabolic pathways.

Background: Alzheimer's disease (AD) is the most well-known reason for disability in persons aged greater than 65 years worldwide. AD influences the part of the brain that controls cognitive and non-cognitive functions. Objective: The study focuses on the screening of natural compounds for the inhibition of AChE and BuChE using a computational methodology. Methods: We performed a docking-based virtual screening utilizing the 3D structure of AChE and BuChE to search for potential inhibitors for AD. In this work, a screened inhibitor Ajmalicine similarity search was carried out against a natural products database (Super Natural II). Lipinski rule of five was carried out and docking studies were performed between ligands and enzyme using ‘Autodock4.2’. Results: wo phytochemical compounds SN00288228 and SN00226692 were predicted for the inhibition of AChE and BuChE, respectively. The docking results revealed Ajmalicine, a prominent natural alkaloid, showing promising inhibitory potential against AChE and BuChE with the binding energy of -9.02 and -8.89 kcal/mole, respectively. However, SN00288228- AChE, and SN00226692-BuChE were found to have binding energy -9.88 and -9.54 kcal/mole, respectively. These selected phytochemical compounds showed better interactions in comparison to Ajmalicine with the target molecule. Conclusion: The current study verifies that SN00288228 and SN00226692 are more capable inhibitors of human AChE and BuChE as compared to Ajmalicine with reference to ΔG values.

Introduction The formula of Chinese medicine, Ding-Zhi-Xiao-Wan (DZXW), has the distinct feature of compatibility therapy, which is attributed to the interactions of multi-herbs. However, the quantification problem caused by the absence of pure reference standards is a bottleneck to clarify the compatibility advantages from the perspective of pharmacokinetics (PKs). Objective This study aimed to develop a putative multiple-reaction monitor (PMRM) strategy for exploring the comparative PKs of DZXW and its single herbs. Methods First, precursor ion and tandem mass spectrometry (MS/MS) chromatograms were obtained via ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight MS (UHPLC-Q-TOF-MS) under different collision energy (CE) values. Then, the two most abundance ions in the MS/MS chromatograms were chosen as product ions, and CE values were selected according to the abundance of the product ion peaks. Next, a PMRM strategy consisting of optimal MRM parameters was constructed. Finally, the established PMRM parameters were imported to UHPLC coupled with triple quadrupole MS (UHPLC-TQ-MS) for quantification. Results The strategy was exemplified by the comparative PK study of DZXW and its single herbs. This strategy could extend the PK scopes of multi-components. The quantitative results displayed substantial variations in PK parameters between DZXW and its single herbs. Conclusion The PK parameters indicated that the DZXW formula could increase the exposure levels of most ingredients and reduce the maximum concentration (Cmax ) of Radix Polygala, indicating that herb compatibility could produce synergistic effects and diminish possible toxic effects. This study provides a viable orientation for the compatibility investigation of traditional Chinese medicine.

Background Inflammation and angiogenesis are the two dominant mechanisms of diabetic retinopathy (DR), which act more as mutual pathways rather than individual processes. However, the underlying mechanism of their interactions is still unclear. Here, we explored the potential crossing points between these pathways and the targeted therapeutic method in rats with DR. Materials and methods Sprague-Dawley rats were randomly assigned to four groups: normal control group, streptozocin (STZ)-induced diabetes mellitus (DM) group, DM+shNC (non-specific negative control shRNA) group, and DM+shNLRP3 group. Silencing the NLR family pyrin domain containing 3 (NLRP3) protein was performed by intravitreal injections of NLRP3-targeted shRNA (shNLRP3) for rats in the DM+shNLRP3 group. All the rats' retinas were collected for further morphological examination and pro-inflammatory and pro-angiogenic cytokine detection. Human retinal endothelial cells (HRECs) were also employed to explore the underlying mechanism. Results NLRP3-targeted shRNA given by intravitreal injection effectively alleviated the retinal histopathological changes in STZ-induced diabetic rats, which reduced the activation of the NLRP3 inflammasome and suppressed the expressions of hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF), and inflammatory cytokines in diabetic rats' retinas. In HRECs, NLRP3 over-expressing plasmid evoked an increase in pro-inflammatory cytokines and VEGF. In addition, YC-1, a HIF-1α inhibitor, could reverse the NLRP3 over-expression-induced VEGF production but not the pro-inflammatory cytokine expressions. Conclusion Our results suggest NLRP3 inflammasome as the potential cross-point between inflammation and pro-angiogenesis in DR and support the effectiveness of NLRP3-targeted shRNA administrated by intravitreal injection in animal models of DR. The protective effect of NLRP3-targeted shRNA may stem from the inhibition of both pro-inflammatory cytokines and HIF-1α/VEGF axis.

It has been 36 years since ISO 4298:1984 was published, yet the role of sodium pyrophosphate in the oxidation of manganese(II) by potassium permanganate in this standard has not been fully recognized. The sample and reagent dosages exceed the actual detection needs, which creates challenges in the test operation and waste liquid treatment. In this study, a more scientific expression for the principle and reaction and the dosage of pyrophosphate in ISO 4298:1984 are discussed. The pyrophosphate ligand is involved in the oxidation of manganese(II) by potassium permanganate, and plays a role in preventing Mn(II) precipitation or Mn(III) dismutation during the whole reaction. The most appropriate amount of saturated sodium pyrophosphate is 150 mL for the test of manganese ores and concentrates with manganese content from 15% to 60%. The method developed by reducing sample size and reagent consumption was evaluated with ten certified reference materials, showing high accuracy and meeting the analytical performance requirements. These results can provide technical support for the revision of ISO 4298:1984. Graphical abstract.

Organic solvent-tolerant proteases have many applications in the synthesis of peptides. In this study, we have developed a low-cost and convenient method to produce highly concentrated organic solvent-tolerant protease. Organic solvent tolerant protease (OSP) gene from Bacillus sphaericus DS11 was cloned and expressed in Bacillus subtilis WB800. The optimum pH of the recombinant protease was 9.0. The optimum temperature of the recombinant protease was 40 °C. The recombinant protease was purified by ethanol with the yield of (87.33%). The yield of OSP enriched by ethanol was higher than that of by Ni-chelating affinity chromatography, which indicated that precipitation of the recombinant OSP with ethanol is a relatively low-cost and fast method for organic solvent -tolerant protease preparation. These results showed that this enzyme could be very useful in different industrial applications.

Aims/Introduction To assess the efficacy and safety of metformin/sitagliptin‐based dual/triple therapy in elderly Chinese patients with type 2 diabetes mellitus. Materials and Methods This subgroup analysis included individuals aged ≥65 years from the STRATEGY study, a two‐stage study in which type 2 diabetes mellitus patients with unsatisfactory glycemic control on metformin were first treated with the dual combination of metformin and sitagliptin for 16 weeks (n = 681), and then, if glycemic control had not been achieved, were treated with a third add‐on oral antihyperglycemic drug for another 24 weeks (n = 291). The efficacy end‐point was change in glycated hemoglobin (HbA1c) in each stage, and the safety end‐point was adverse events with a focus on hypoglycemia. Results At week 16, the change in HbA1c was −0.81% from baseline, and the percentages of patients who achieved HbA1c targets of, Our findings provide evidence that metformin/sitagliptin‐based dual therapy can significantly improve glycemic control in elderly Chinese type 2 diabetes mellitus patients with unsatisfactory glycemic control on metformin, even in those with a long duration of diabetes, mild chronic kidney disease or overweight/obesity. The addition of a third add‐on agent, including glimepiride, gliclazide, repaglinide or acarbose, was relatively well‐tolerated, with a neutral effect on weight, and can further improve glycemic control in elderly type 2 diabetes mellitus patients.

The chitin deacetylase produced by marine strain Nitratireductor aquimarinus MCDA3-3 (NaCDA) was purified by using ammonium sulfate precipitation, Q Sepharose, and Superdex column chromatography. The purified NaCDA showed 75-fold purity, 50 U/mg specific activity with 28.5% yield. The purified CDA molecular weight was about 36 kDa. The temperature and pH of the purified enzyme were suiting at 30 °C and 8.0, respectively. The NaCDA was highly stable for a wide range of temperature 4 °C–25 °C and pH 6.0–9.0. Besides, increased enzyme activity was observed by introducing metal ions mainly Sr2+, Mg2+, and Na+. The enzyme was founded inhibited by Co2+, Ba2+, and EDTA at the value of 1 mM concentrations. On the other hand, NaCDA was shown an active activity behavior toward glycol chitin and chitin oligomers with a degree of polymerization more than four, any polymer below the chain such as trimer and dimer significantly reduce in the activity rate, and inactive with N-acetylglucosamine. Interestingly, NaCDA showed a high activity rate against insoluble chitins and converting acetyls to deacetylated. The reduction of acetyls from 56.26% and 22.88% of acetyl groups from ɑ-chitin and β-chitin, respectively. Hence, the NaCDA would be applicable in production chitosan toward mass production.

Serpinb6b is a novel member of Serpinb family and found in germ and somatic cells of mouse gonads, but its physiological function in uterine decidualization remains unclear. The present study revealed that abundant Serpinb6b was noted in decidual cells, and advanced the proliferation and differentiation of stromal cells, indicating a creative role of Serpinb6b in uterine decidualization. Further analysis found that Serpinb6b modulated the expression of Mmp2 and Mmp9. Meanwhile, Serpinb6b was identified as a target of Bmp2 regulation in stromal differentiation. Treatment with rBmp2 resulted in an accumulation of intracellular cAMP level whose function in this differentiation program was mediated by Serpinb6b. Addition of PKA inhibitor H89 impeded the Bmp2 induction of Serpinb6b, whereas 8‐Br‐cAMP rescued the defect of Serpinb6b expression elicited by Bmp2 knock‐down. Attenuation of Serpinb6b greatly reduced the induction of constitutive Wnt4 activation on stromal cell differentiation. By contrast, overexpression of Serpinb6b prevented this inhibition of differentiation process by Wnt4 siRNA. Moreover, blockage of Wnt4 abrogated the up‐regulation of cAMP on Serpinb6b. Collectively, Serpinb6b mediates uterine decidualization via Mmp2/9 in response to Bmp2/cAMP/PKA/Wnt4 pathway.

It has long been predicted that oscillatory behavior exists in reactivity as a function of collision energy for heavy-light-heavy (HLH) chemical reactions in which a light atom is transferred between two heavy atoms or groups of atoms, but direct observation of such a behavior in bimolecular reactions remains a challenge. Here we report a joint theoretical and crossed-molecular-beam study on the Cl + CH4 → HCl + CH3 reaction. A distinctive peak at a collision energy of 0.15 eV for the CH3(v = 0) product was experimentally detected in the backward scattering direction. Detailed quantum-dynamics calculations on a highly accurate potential energy surface revealed that this feature originates from the reactivity oscillation in this HLH polyatomic reaction. We anticipate that such reactivity oscillations exist in many HLH reactions involving polyatomic reagents.

In this work, UV irradiation was introduced to promote the etching process of ZnO nanorods towards nanotubes. The ZnO nanorods with different diameters were grown on Au cylindrical spirals via hydrothermal decomposition, and then in the presence of UV irradiation, they were chemically etched to nanotubes. The results indicate that the ZnO nanorods with the larger diameter are easily etched to nanotubes due to enough oxygen vacancies (a kind of defect) within the centre parts, but the ZnO nanorods with the smaller diameter are difficult to form tubular structures because of the insufficient oxygen vacancies. With the introduction of UV irradiation, more oxygen vacancies are produced in the ZnO nanorods and hence, the nanorods with the larger diameter are etched more quickly, and the nanorods with the smaller diameter are etched more easily. Additionally, the UV light with the shorter wavelength is more beneficial for the formation of the ZnO nanotubes than the one with the longer wavelength. The results benefit the wet etching of ZnO nanorods towards nanotubes, especially those with smaller diameters.

Enrichment of trace bioactive constituents and metabolites from complex biological samples is challenging. This study presented a one-pot synthesis of magnetic polydopamine nanoparticles (Fe3O4@SiO2@PDA NPs) with multiple recognition sites for the magnetic dispersive solid-phase extraction (MDSPE) of ginsenosides from rat plasma treated with white ginseng. The extracted ginsenosides were characterized by combining an ultra-high-performance liquid chromatography coupled to a high-resolution mass spectrometry with supplemental UNIFI libraries. Response surface methodology was statistically used to optimize the extraction procedure of the ginsenosides. The reusability of Fe3O4@SiO2@PDA NPs was also examined and the results showed that the recovery rate exceeded 80% after recycling 6 times. Furthermore, the proposed method showed greater enrichment efficiency and could rapidly determine and characterize 23 ginsenoside prototypes and metabolites from plasma. In comparison, conventional methanol method can only detect 8 ginsenosides from the same plasma samples. The proposed approach can provide methodological reference for the trace determination and characterization of different bioactive ingredients and metabolites of traditional Chinese medicines and food., Graphical abstract Image 1, Highlights • The Fe3O4@SiO2@PDA NPs were synthesized through one-pot method. • The RSM was designed to promote the extraction of trace active ingredients. • The MDSPE, UPLC-MS and UNIFI software were integrated into an analytical platform. • The synergetic strategy was applied to enrich ginsenosides from rat plasma. • The synergetic strategy provided an easy, rapid and sensitive method for analytes.

As an important Chinese medicine decoction, Wu-tou decoction has been used to treat rheumatic arthritis for more than a thousand years. We previously reported that the Wu-tou decoction could change the urinary and serum metabolites in adjuvant-induced arthritis rats significantly. The purpose of this research was to confirm the potential biomarkers obtained by previous non-targeted metabolomics study through quantitative analysis by liqui chromatography with tandem mass spectrometry, in the meantime, to further study the effective material basis of Wu-tou decoction. Firstly, the important compounds in the tryptophan metabolism pathway, the arginine and proline metabolism pathway, the amino acid metabolism pathway, the tricarboxylic acid cycle, the vitamin B6 metabolism pathway, and the phenylalanine metabolism pathway, which were identified as potential biomarkers in previous study, were selected for quantitative analysis. Then the linearity, limit of detection, limit of quantification, selectivity, accuracy, precision, stability, recovery, and matrix effect of the quantitative method were examined. Finally, ten and eighteen metabolites were quantitatively analyzed in the serum and urine, respectively. The results showed that seven out of ten serum potential biomarkers and ten out of eighteen urine potential biomarkers were confirmed as real biomarkers. This research provides a powerful reference for the study on effective material basis of Wu-tou decoction.

Citri Exocarpium Rubrum (CER) and Citri Reticulatae Pericarpium (CRP) are used as common functional foods and traditional Chinese medicine (TCM). As different parts of the same fruit, CER and CRP have different effects in clinical applications. However, they are commonly confused due to the similarity of the chemical compounds and a lack of scientific method to distinguish them in the finished product. In this study, an ultra-high performance liquid chromatography-diode array detector (UHPLC-DAD) technique and an ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF MS) method were employed to generate the characteristic fingerprint under the optimum analytical conditions. Principal component analysis (PCA) and orthogonal partial least squares discrimination analysis (OPLS-DA) were applied to represent different chemical markers for CER and CRP. 44 potential markers including 15 polymethoxylated flavanones (PMFs), 5 flavone-C-glycosides, 6 flavanone-O-glycosides, 3 flavonoid-O-glycosides, 8 organic acids, 5 limonoids and 2 alkaloids were successfully identified by using UNIFI software. The heat map showed that there were significant differences in the CER and CRP samples. Furthermore, 12 potential markers were screened out by characteristic fingerprint and UHPLC-Q-TOF MS methods and were analyzed by quantitative analysis of multicomponents by a single marker (QAMS) method. Finally, a prediction model based on the discovered chemical markers was established for discrimination between CER and CRP. Using these markers can significantly distinguish the unknown processed products of CER and CRP. In conclusion, an effective way to quickly and easily distinguish CER and CRP was successfully established based on the characteristic fingerprint and UHPLC-Q-TOF MS. It could also be a new strategy for analysis of different processed products of the same plant source.

The environmental behavior and toxicity of nanomaterials can be significantly affected by the ubiquitous natural organic matter (NOM) of aquatic environments. However, general mechanisms governing the effects of different organic matter samples on nanomaterials remain elusive at present. Thus, the environmental behavior and toxicity of organic matter with different components or conformation, such as humic acid (HA), bovine serum albumin (BSA) and denatured BSA (DB), on five nanomaterials (i.e., Ag nanoparticles (NPs), polystyrene nanoparticles (PS NPs), TiO2 NPs, nano-zerovalent iron (NZVI), and multiwall carbon nanotubes (MWCNTs)) to Tetrahymena thermophila (T. thermophila) were studied. BSA and DB were found to significantly increase (up to 99%) the bioaccumulation of nanomaterials (Ag NPs and PS NPs) in T. thermophila by specific recognition. This increase was lower in DB-coated nanomaterials, compared to BSA-coated nanomaterials, revealing the importance of protein conformation in the specific recognition process. The toxicity results revealed that different organic matter samples mostly affected Ag NPs and NZVI (toxicity by dissolved metal ions) by altering their biological uptake. In the case of nanomaterials whose toxicity was dominated by suspended particles, the organic matter could alter the toxicity of these materials by changing their surface properties and their state in environmental media, and by scavenging free radicals, instead of altering the bioaccumulation process. In general, the toxicity of HA-coated nanomaterials was significantly lower than those of DB- or BSA-coated nanomaterials. Our results suggest that it is essential to consider the type and characteristics of the organic matter when evaluating the toxicity of NPs, since this represents a more realistic state of exposure.

Leaves of Acanthopanax senticosus (Rupr. et Maxim.) Harms (ASL) have revealed significant biological activity in the treatment of ischemic stroke diseases. However, there was no in-depth study of the therapeutic material basis and effect of ASL from the pharmacokinetics-pharmacodynamics (PK-PD) analysis level. In this study, a method based on microdialysis coupled with ultra-performance liquid chromatography combined with triple quadruple mass spectrometry (MD-UPLC-QQQ-MS) was established to simultaneously and continuously collect and quantify the active compounds and endogenous neuroactive substances related to therapeutic effect in plasma and hippocampus of fully awake ischemic stroke rats. The acquired data were analyzed by the PK-PD analysis method. It was found that hyperoside, quercitrin, quercetin, and caffeic acid could pass through the blood-brain barrier, and quercetin needed a longer intake time than quercitrin and hyperoside, but the passage rate was higher. The exposure of the four compounds in the hippocampus affected the contents of seven neuroactive substances in different ways and was depicted graphically (concentration-time effect). In addition, the study found that the brain index and brain water content of ischemic stroke rats were significantly reduced after the oral administration of ASL. ASL observably regulated the content or activity of six important biochemical indexes in rats. On the one hand, this study verified that ASL could regulate ischemic stroke in many aspects. On the other hand, a visualized method to express the relationship between pharmacokinetics and pharmacodynamics in the hippocampus of cerebral ischemic areas was established. This research gives a hand to the study on the therapeutic material basis and effect of traditional Chinese medicine mechanism.

Nanoliposome is a useful dosage form to increase solubility and absorption of simvastatin (SMV), and consequently improves its therapeutic effects. However, in vivo toxicity of SMV could also be elevated accompanied by the absorption enhancement, which is a decisive factor for the clinical application of SMV nanoliposome (SMV-Lipo), but has not been studied systematically and reported so far. In this study, organ toxicity of SMV-Lipo was evaluated in mice in the presence and absence of isoproterenol and compared to those of free SMV. Results demonstrated that compared to free SMV, the SMV-Lipo administrated at an equal dose of 25 mg/kg/d led to severe myocardiotoxicity, hepatotoxicity at baseline and more pronounced liver injury with elevation of alanine aminotransferase. In addition, muscular adverse effect was also observed in SMV-Lipo treated group but not in SMV group. Pharmacokinetic studies revealed that compared to free SMV, the SMV-Lipo administration significantly improved the plasma SMV concentration, and the oral bioavailability was 6.5 times of free SMV. Notably, when the dosage of free SMV increased to 50 mg/kg/d, yielding the comparable plasma concentration as SMV-Lipo given at 25 mg/kg/d, the myocardiotoxicity was observed in free SMV treated mice as well, which further confirmed that the enhanced absorption of SMV by the nanoliposomal formulation resulted in more severe myocardiotoxicity than the equal dose of free SMV., Graphical abstract When given at the same dosage and administration regimen, the SMV-Lipo presented significant myocardiotoxicity due to the significant enhancement of the plasma SMV concentration by nanoliposomes.Image, graphical abstract