ABSTRACT
Plant roots encounter numerous pathogenic microbes that often cause devastating diseases. One such pathogen, Plasmodiophora brassicae (Pb), causes clubroot disease and severe yield losses on cruciferous crops worldwide. Here, we report the isolation and characterization of WeiTsing (WTS), a broad-spectrum clubroot resistance gene from Arabidopsis. WTS is transcriptionally activated in the pericycle upon Pb infection to prevent pathogen colonization in the stele. Brassica napus carrying the WTS transgene displayed strong resistance to Pb. WTS encodes a small protein localized in the endoplasmic reticulum (ER), and its expression in plants induces immune responses. The cryoelectron microscopy (cryo-EM) structure of WTS revealed a previously unknown pentameric architecture with a central pore. Electrophysiology analyses demonstrated that WTS is a calcium-permeable cation-selective channel. Structure-guided mutagenesis indicated that channel activity is strictly required for triggering defenses. The findings uncover an ion channel analogous to resistosomes that triggers immune signaling in the pericycle.
Subject(s)
Brassica napus , Plasmodiophorida , Cryoelectron Microscopy , Lead , Brassica napus/genetics , Plasmodiophorida/physiology , Ion Channels , Plant DiseasesABSTRACT
An erratum is presented to modify a calculating error in our published manuscript ["High-power 970â nm semiconductor disk laser" Opt. Express31, 43963 (2023)10.1364/OE.506462 [CrossRef]]. All results throughout the manuscript and its conclusions are unaffected by this correction and remain valid.
ABSTRACT
Semiconductor disk lasers (SDLs) have emerged at the frontier of laser technologies. Here, the chip design, packaging process, resonator, pumping strategy, etc. are optimized for the performance improvement of a 970â nm SDL. After optimization, a power of 70.3 W is attained under continuous wave (CW) operation, and the corresponding thermal resistance is around 0.49â K/W. The laser is highly efficient with a maximum slope efficiency of 58.2% and the pump threshold is only around 1.83â kW/cm2. Furthermore, the emission performances under quasi-continuous wave (QCW) pumping are also explored. Setting the duty cycle to about 11%, the chips can output a peak power of 138 W without thermal rollover, and the single pulse energy can reach about 13.6 mJ. As far as we know, they are the best results in terms of power/energy in this wavelength SDL. These explorations may help to understand the thermal characteristics in high-power SDLs and may also be regarded as an extension and enrichment of the earlier works on this topic.
ABSTRACT
Background and Objective: Our previous study demonstrated that consistent treatment of oral cilostazol was effective in reducing levels of painful peripheral neuropathy in streptozotocin-induced type I diabetic rats. As diabetic neuropathy is characterized by hyperglycemia-induced nerve damage in the periphery, this study aims to examine the neuropathology as well as the effects of cilostazol treatments on the integrity of peripheral small nerve fibers in type I diabetic rats. Materials and Methods: A total of ninety adult male Sprague-Dawley rats were divided into the following groups: (1) naïve (control) group; (2) diabetic rats (DM) group for 8 weeks; DM rats receiving either (3) 10 mg/kg oral cilostazol (Cilo10), (4) 30 mg/kg oral cilostazol (Cilo30), or (5) 100 mg/kg oral cilostazol (Cilo100) for 6 weeks. Pain tolerance thresholds of hind paws toward thermal and mechanical stimuli were assessed. Expressions of PGP9.5, P2X3, CGRP, and TRPV-1 targeting afferent nerve fibers in hind paw skin and glial cells in the spinal dorsal horn were examined via immunohistochemistry and immunofluorescence. Results: Oral cilostazol ameliorated the symptoms of mechanical allodynia but not thermal analgesia in DM rats. Significant reductions in PGP9.5-, P2X3-, CGRP, and TRPV-1-labeled penetrating nerve fibers in the epidermal layer indicated denervation of sensory nerves in the hind paw epidermis of DM rats. Denervation significantly improved in groups that received Cilo30 and Cilo100 in a dose-dependent manner. Cilostazol administration also suppressed microglial hyperactivation and increased astrocyte expressions in spinal dorsal horns. Conclusions: Oral cilostazol ameliorated hyperglycemia-induced peripheral small nerve fiber damage in the periphery of diabetic rats and effectively mitigated diabetic neuropathic pain via a central sensitization mechanism. Our findings present cilostazol not only as an effective option for managing symptoms of neuropathy but also for deterring the development of diabetic neuropathy in the early phase of type I diabetes.
Subject(s)
Diabetes Mellitus, Experimental , Diabetes Mellitus, Type 1 , Diabetic Neuropathies , Hyperglycemia , Rats , Male , Animals , Cilostazol/therapeutic use , Cilostazol/pharmacology , Diabetic Neuropathies/drug therapy , Rats, Sprague-Dawley , Streptozocin/adverse effects , Diabetes Mellitus, Experimental/complications , Diabetes Mellitus, Experimental/chemically induced , Calcitonin Gene-Related Peptide/adverse effects , Calcitonin Gene-Related Peptide/analysis , Sciatic Nerve/pathology , Hyperalgesia/drug therapy , Hyperalgesia/etiology , Hyperalgesia/metabolism , DenervationABSTRACT
BACKGROUND: The mainstream facilitation of one-lung ventilation is using double-lumen endobronchial tubes. However, it is more difficult to be positioned properly and more likely to cause airway injuries. How to place double-lumen endobronchial tubes rapidly and correctly is important for thoracic anesthesiologists. METHODS: One hundred eight patients with an American Society of Anesthesiologists physical status of I to III were 20 years of age or over, and required one-lung ventilation for thoracic surgery. They were randomly assigned to the conventional technique group (n = 36), the flexible fiberoptic bronchoscopy group (n = 36), or the Trachway® flexible stylet group (n = 36). The primary endpoint was the time needed for intubation. T1, the time from the tip of the blade passing between the patient's lips to identification of the vocal cords; and T2, the time from identification of the vocal cords to the bronchial lumen was in the correct position. RESULTS: T1 had no significant difference between groups, but T2 was significantly shorter in the Trachway® flexible stylet group (p < 0.0001) and longer in the conventional technique group (p < 0.0001). CONCLUSIONS: Using Trachway® flexible stylet for correct placement of double-lumen endobronchial tubes not only significantly shortened the intubation time, but also reduced incidence of carinal injuries. It is an alternative, and a choice with good safety. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT02364622, 18/02/2015, Retrospectively registered.
Subject(s)
Intubation, Intratracheal , One-Lung Ventilation , Bronchi , Bronchoscopy/methods , Humans , Intubation, Intratracheal/methods , Prospective StudiesABSTRACT
Background and Objectives: The anterolateral thigh (ALT) flap is widely used in head and neck reconstruction, but the postoperative thigh sensory function lacks sufficient evaluation. The present study reports the postsurgical pain and cancer-related quality of life (QoL) in different stages of oral cancer patients receiving anterolateral thigh (ALT) flap reconstruction. Materials and Methods: Patients were subgrouped into postoperative early-, mid-, and late-recovery stages (postoperative 0.5-1 years, 1-2 years, and above 2 years) according to the time point of assessment. The QoL was examined using the EORTC C-30. Postsurgical donor and receipt site pain was evaluated through subjective reports and sensory tests. Results: Ninety-four patients were included in the final analysis. The functional and global health-related QoL significantly improved with time after surgery. However, spontaneous pain was reported in 57.7%, 72.3%, and 42% of patients in early-, mid-, and late-recovery stages, mainly in donor sites rather than in receipt sites. The highest incidence of donor site pain after ALT flap reconstruction in oral cancer surgery was in the mid-recovery stage but remained high in the late-recovery stage (56.8% and 36.7%, respectively). Conclusions: The postsurgical pain in the donor site might persist to or exhibit delayed onset one to two years postoperatively but is much improved after postoperatively two years later. A longer postsurgical follow-up for over two years for pain and sensory dysfunction is indicated.
Subject(s)
Free Tissue Flaps , Mouth Neoplasms , Humans , Mouth Neoplasms/complications , Mouth Neoplasms/surgery , Pain, Postoperative/etiology , Quality of Life , Thigh/surgeryABSTRACT
Photoelectrochemical (PEC) sensors are relatively new sensing platforms with high detection sensitivity and low cost. However, the current PEC biosensors dependent on ultraviolet or visible light as the exciting resource cause injuries to biological samples and systems, which restrains the applications in complicated matrixes. Herein, a near-infrared light (NIR)-initiated PEC biosensor based on NaYF4:Yb,Tm@NaYF4@TiO2@CdS (csUCNRs@TiO2@CdS) was constructed for sensitive detection of acute myocardial infarction (AMI)-related miRNA-133a in an immobilization-free format coupled with a hybridization chain reaction and a redox circle signal amplification strategy. A low-energy 980 nm NIR incident laser was converted to 300-480 nm light to excite the adjacent TiO2@CdS photosensitive shell to generate photocurrent by NaYF4:Yb,Tm@NaYF4 upconversion nanorods. Also, magnetic beads were employed for the homogeneous determination of target miRNA-133a to reduce the recognition steric hindrance and improve the detection sensitivity. The photocurrent response was positively correlated with the level of ascorbic acid as the energy donor to consume photoacoustic holes produced on the surface of csUCNRs@TiO2@CdS, which was generated by alkaline phosphatase catalyzation and regenerated by tris(2-carboxyethyl) phosphine reduction upon the appearance of miRNA-133a. Exerting a NIR-light-driven and immobilization-free strategy, the as-constructed biosensor displayed linearly sensitive and selective determination of miRNA-133a with a detection limit of 36.12 aM. More significantly, the assay method provided a new concept of the PEC sensing strategy driven by NIR light to detect diverse biomarkers with pronounced sensitivity, light stability, and low photodamage.
Subject(s)
Biosensing Techniques , MicroRNAs , Nanotubes , Electrochemical Techniques , Infrared Rays , Limit of DetectionABSTRACT
BACKGROUND: Preterm neonates are at higher risk of developing inguinal hernia, and have an increased risk of perioperative adverse events. Laryngeal mask airway (LMA) is claimed to be associated to decreasing perioperative respiratory complications compared to endotracheal tube (ETT) in infants under one year of age receiving minor surgery; thus, we conducted a retrospective survey in former preterm neonates below 5000 g to compare the respiratory complications between LMA and ETT in general anesthesia for inguinal hernia surgeries. METHODS: The inclusion criteria were: gestational age at birth under 37 weeks, body weight at surgery below 5000 g, and receiving scheduled inguinal hernia repair under general anesthesia with LMA or ETT. Infants who were dependent on mechanical ventilation preoperatively were excluded. The postoperative respiratory complications including delayed extubation, re-intubation, and apnea within postoperative 24 h were compared between groups. RESULTS: From July 2014 to December 2017, 72 neonates were enrolled into final analysis. There were 57 neonates managed with LMA, and only 15 neonates intubated with ETT during the study period. The gestational age at birth and post-menstrual age at surgery showed no significant difference between groups, although in the ETT group, the body weight at birth and at surgery were lower, and more infants had history of severe respiratory distress syndrome and had received oxygen therapy within two weeks prior to surgery. Surprisingly, none one of the infants developed delayed extubation, re-intubation, or postoperative apnea in the LMA group. In the ETT group, 40 percent of the neonates could not be successfully extubated in the operation theater. CONCLUSION: In preterm neonates, even in those younger than 52 weeks post-menstrual age who undergoing inguinal hernia repair in their early infancy, LMA appears feasible and safe as the airway device during general anesthesia in specific patient group. However, anesthesiologist might prefer ETT rather than LMA in some complex situation. In neonates with lower body weight at birth and at surgery, and with a history of severe RDS and oxygen-dependence, further prospective study is required.
Subject(s)
Hernia, Inguinal/surgery , Intubation, Intratracheal/methods , Laryngeal Masks , Postoperative Complications/epidemiology , Airway Extubation/statistics & numerical data , Anesthesia, General/methods , Birth Weight , Female , Humans , Infant, Newborn , Infant, Premature , Male , Retrospective StudiesABSTRACT
MicroRNAs (miRNAs) are short, endogenous, noncoding RNAs that play critical roles in physiologic and pathologic processes and are vital biomarkers for several disease diagnostics and therapeutics. Therefore, rapid, low-cost, sensitive, and selective detection of miRNAs is of paramount importance and has aroused increasing attention in the field of medical research. Among the various reported miRNA sensors, devices based on graphene and its derivatives, which form functional supramolecular nanoassemblies of π-conjugated molecules, have been revealed to have great potential due to their extraordinary electrical, chemical, optical, mechanical, and structural properties. This Review critically and comprehensively summarizes the recent progress in miRNA detection based on graphene and its derivative materials, with an emphasis on i) the underlying working principles of these types of sensors, and the unique roles and advantages of graphene materials; ii) state-of-the-art protocols recently developed for high-performance miRNA sensing, including representative examples; and iii) perspectives and current challenges for graphene sensors. This Review intends to provide readers with a deep understanding of the design and future of miRNA detection devices.
Subject(s)
Graphite/chemistry , MicroRNAs/metabolism , Animals , Biosensing Techniques/methods , Electrochemical Techniques/methods , Humans , MicroRNAs/chemistry , Nanostructures/chemistryABSTRACT
Pattern-recognition receptors (PRRs), which consist of receptor kinases (RKs) and receptor-like proteins, sense microbe- and host-derived molecular patterns associated with pathogen infection to trigger immune responses in plants. Several kinases of the 46-member Arabidopsis (Arabidopsis thaliana) receptor-like cytoplasmic kinase (RLCK) subfamily VII play important roles in pattern-triggered immunity, but it is unclear whether different RLCK VII members act specifically or redundantly in immune signaling. Here, we constructed nine higher order mutants of this subfamily (named rlck vii-1 to rlck vii-9) and systematically characterized their immune phenotypes. The mutants rlck vii-5, -7, and -8 had compromised reactive oxygen species production in response to all patterns tested, indicating that the corresponding members are broadly required for the signaling of multiple PRRs. However, rlck vii-4 was defective specifically in chitin-induced reactive oxygen species production, suggesting that RCLK VII-4 members mediate the signaling of specific PRRs. Furthermore, RLCK VII-4 members were required for the chitin-triggered activation of MAPK, demonstrating that these kinases link a PRR to MAPK activation. Moreover, we found that RLCK VII-6 and -8 also were required for RK-mediated root growth. Together, these results show that numerous RLCK VII members are involved in pattern-triggered immune signaling and uncover both common and specific roles of these kinases in plant development and immunity mediated by various RKs.
Subject(s)
Arabidopsis Proteins/immunology , Arabidopsis/physiology , Plant Immunity , Protein Kinases/immunology , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Chitin/metabolism , Disease Resistance/immunology , Gene Expression Regulation, Plant/immunology , Mitogen-Activated Protein Kinases/immunology , Mitogen-Activated Protein Kinases/metabolism , Mutation , Phylogeny , Plant Diseases/immunology , Plant Immunity/genetics , Plants, Genetically Modified , Protein Kinases/genetics , Protein Kinases/metabolism , Pseudomonas syringae/pathogenicity , Reactive Oxygen Species/immunology , Reactive Oxygen Species/metabolism , Receptors, Pattern Recognition/genetics , Receptors, Pattern Recognition/metabolism , Signal TransductionABSTRACT
The superoxide anion (O2Ë-) plays a crucial role in several physiological processes and many human diseases. Developing new methods for O2Ë- detection in biological systems is very important. A FRET-based two-photon (TP) fluorescent probe with a ratiometric signal, TFR-O, was developed. A naphthalene derivative based TP fluorescent group was selected as the energy donor group, and a rhodol fluorescent group was chosen as the energy acceptor; the trifluoromethanesulfonate group was chosen as the recognition moiety. After reacting with O2Ë-, the recognition moiety was removed and the fluorophore was released, leading to a fluorescence intensity decrease at the wavelength of 425 nm and a significant enhancement of the fluorescence intensity at 550 nm. The fluorescence intensity ratio between 550 and 425 nm (I550/I425) varied from 0.15 to 6.72, with the O2Ë- concentration increasing from 0 to 50 µM. The detection limit of the TFR-O was 83 nM. Moreover, TFR-O was applied for detecting and imaging O2Ë- in cells and liver tissues.
Subject(s)
Fluoresceins/chemistry , Fluorescent Dyes/chemistry , Mesylates/chemistry , Naphthalenes/chemistry , Superoxides/analysis , Animals , Fluoresceins/chemical synthesis , Fluoresceins/radiation effects , Fluoresceins/toxicity , Fluorescence , Fluorescence Resonance Energy Transfer/methods , Fluorescent Dyes/chemical synthesis , Fluorescent Dyes/radiation effects , Fluorescent Dyes/toxicity , Limit of Detection , Liver/metabolism , Mesylates/chemical synthesis , Mesylates/radiation effects , Mesylates/toxicity , Mice , Naphthalenes/chemical synthesis , Naphthalenes/radiation effects , Naphthalenes/toxicity , Photons , RAW 264.7 Cells , Superoxides/metabolismABSTRACT
The lumen degradation of LED lamps undergoing an accelerated aging test is investigated. The entire LED lamp is divided into three subsystems, namely, driver, lampshade, and LED light source. The parameters of output power [Watts (W)], transmittance (%), and lumen flux (lm) are adopted in the analysis of the degradation of the driver, lampshade, and LED light source, respectively. Two groups of LED lamps are aged under the ambient temperatures of 25°C and 85°C, respectively, with the aging time of 2000 h. The lumen degradation of the lamps is from 3.8% to 4.9% for the group under a temperature of 25°C and from 10.6% to 12.7% for the group under a temperature of 85°C. The LED light source is the most aggressive part of the three subsystems, which accounts for 70.5% of the lumen degradation of the LED lamp on average. The lampshade is the second degradation source, which causes 21.5% of the total amount on average. The driver is the third degradation source, which causes 6.5% under 25°C and 2.8% under 85°C of the total amount on average.
ABSTRACT
Small ubiquitin-like modifier (SUMO1-3) conjugation plays a critical role in embryogenesis. Embryos deficient in the SUMO-conjugating enzyme Ubc9 die at the early postimplantation stage. Sumo1(-/-) mice are viable, as SUMO2/3 can compensate for most SUMO1 functions. To uncover the role of SUMO2/3 in embryogenesis, we generated Sumo2- and Sumo3-null mutant mice. Here, we report that Sumo3(-/-) mice were viable, while Sumo2(-/-) embryos exhibited severe developmental delay and died at approximately embryonic day 10.5 (E10.5). We also provide evidence that SUMO2 is the predominantly expressed SUMO isoform. Furthermore, although Sumo2(+/-) and Sumo2(+/-);Sumo3(+/-) mice lacked any overt phenotype, only 2 Sumo2(+/-);Sumo3(-/-) mice were found at birth in 35 litters after crossing Sumo2(+/-);Sumo3(+/-) with Sumo3(-/-) mice, and these rare mice were considerably smaller than littermates of the other genotypes. Thus, our findings suggest that expression levels and not functional differences between SUMO2 and SUMO3 are critical for normal embryogenesis.
Subject(s)
Embryonic Development , Small Ubiquitin-Related Modifier Proteins/genetics , Ubiquitins/genetics , Animals , Female , Gene Expression , Genes, Essential , Mice, Inbred C57BL , Mice, Knockout , Small Ubiquitin-Related Modifier Proteins/metabolism , Ubiquitins/metabolismABSTRACT
OBJECTIVE: To investigate whether rs12731181 (AâG) interrupted miR-590-3p-mediated suppression of the prostaglandin F2α receptor (FP) and whether it is associated with essential hypertension in the Chinese population. APPROACH AND RESULTS: We found that miR-590-3p regulates human FP gene expression by binding to its 3'-untranslated region. rs12731181 (AâG) altered the binding affinity between miR-590-3p and its FP 3'-untranslated region target, thus reducing the suppression of FP expression, which, in turn, enhanced FP receptor-mediated contractility of vascular smooth muscle cells. Overexpression of FP augmented vascular tone and elevated blood pressure in mice. An association study was performed to analyze the relationship between the FP gene and essential hypertension in the Han Chinese population. The results indicated that the rs12731181 G allele was associated with susceptibility to essential hypertension. Carriers of the AG genotype exhibited significantly higher blood pressure than those of the AA genotype. FP gene expression was significantly higher in human peripheral leukocytes from individuals with the AG genotype than that in leukocytes from individuals with the AA genotype. CONCLUSIONS: rs12731181 in the seed region of the miR-590-3p target site is associated with increased risk of essential hypertension and represents a new paradigm for FP involvement in blood pressure regulation.
Subject(s)
Asian People/genetics , Hypertension/genetics , MicroRNAs/genetics , Receptors, Prostaglandin/genetics , 3' Untranslated Regions , Animals , Binding Sites , China/ethnology , Essential Hypertension , Genetic Predisposition to Disease , Humans , Mice , Polymorphism, Single Nucleotide , Transcription, GeneticABSTRACT
Xenon flash lamp is used by most analytical instruments and medical equipments as light sources whose spectral characteristics exert a tremendous influence on the property of instruments and medical equipments. In order to study the effects of pulsed Xenon discharge circuit parameters and geometric parameters on the spectral characteristics, theoretical analysis has been conducted to Xenon lamp emitting process with the gas discharge theory. The spectroscopic detection system has been designed to conduct experimental detection to the spectral characteristics of pulsed Xenon lamp with different parameters. The experimental results show that the emission spectra of Xenon flash lamp contain both line and continuous radiation. The line spectrum results from the electron energy gap transition, while the cw radiation related to ionic recombination and bremsstrahlung. The relative spectral intensity increases linearly with the discharge voltage, and has a positive correlation with storage capacitor. When low discharge voltage, the shorter the arc length, the higher the relative spectral intensity .while the discharge voltage becomes higher, the result is reversed. The work in this paper has great significance for xenon flashlamp on the parameters selection and industrial application.
ABSTRACT
To predict the mechanism of liver injury induced by Genkwa Flos, we investigated the effect of chloroform extract on UGTs and UGT1A1 activities of the liver microsomes in rat and human. In the present study, 4-nitrophenol(4-NP) and ß-estradiol were elected as substrates to determine activities of UGTs and UGT1A1 by UV and HPLC. The results showed that there were 1.00% of apigenin, 6.40% of hydroxygenkwanin and 18.38% of genkwanin in chloroform extract; and total diterpene mass fraction was 31.40%. Compared with the control group, chloroform extract could significantly inhibit the activity of UGTs in rat liver microsomes(RLM) system, while the inhibitory effect was not obvious in human liver microsomes(HLM) system. UGT1A1 activity was inhibited by chloroform extract in rat liver microsomes and human liver microsomes (based on genkwanin, IC50=8.76, 10.36 µmolâ¢L⻹). The inhibition types were non-competitive inhibition(RLM) and uncompetitive inhibition(HLM). In conclusion, the results indicated that chloroform extract showed different inhibitory effects on UGTs and UGT1A1 activity, which may be one of the mechanisms of liver injury induced by Genkwa Flos.
Subject(s)
Daphne/chemistry , Drugs, Chinese Herbal/pharmacology , Glucuronosyltransferase/metabolism , Microsomes, Liver/drug effects , Plant Extracts/pharmacology , Animals , Apigenin/analysis , Chromatography, High Pressure Liquid , Estradiol , Flavones/analysis , Flavonoids/analysis , Glucuronosyltransferase/antagonists & inhibitors , Humans , Microsomes, Liver/enzymology , Nitrophenols , RatsABSTRACT
In the present study, the effects of six Coptidis alkaloids (berberine, epiberberine, coptisine, jatrorrhizine, palmatine and magnoflorine) on liver microsomes UGTs and UGT1A1 activities in rats and mice were investigated in vitro and in vivo to study the mechanism of metabolic drug-drug interactions of Coptidis Rhizoma with other drugs. In vitro rat and mice liver microsomal incubation systems combined with UDPGA were applied, as well as mice liver microsomes after administration of six Coptidis alkaloids. 4-Nitrophenol and ß-estradiol were selected as substrates to determine activities of UGTs and UGT1A1 by UV and HPLC, respectively. According to the in vitro rat study, berberine, epiberberine, coptisine and jatrorrhizine significantly inhibited rat liver microsome UGTs activity, particularly epiberberine showed the strongest inhibition. UGT1A1 activity was lowly inhibited by jatrorrhizine, with IC50 at about 227 µmolâ¢L⻹, whereas coptisine and magnoflorine significantly activated UGT1A1. According to the in vitro mice study, berberine, coptisine, jatrorrhizine and palmatine significantly inhibited mice liver microsome UGTs activity, and the six alkaloids all significantly activated UGT1A1. According to the in vivo mice study, UGTs activity was significantly activated only in berberine group, while UGT1A1 activity was significantly activated only in jatrorrhizine group. In conclusion, the effects of Coptidis alkaloids on UGT activity showed significant differences in species and between in vitro and in vivo. Meanwhile, the changes in structures of Coptidis alkaloids also have a big impact on UGT activity, which may be one of the causes for the drug-drug interactions between Coptidis Rhizoma and other drugs.
Subject(s)
Alkaloids/administration & dosage , Coptis/chemistry , Drugs, Chinese Herbal/administration & dosage , Glucuronosyltransferase/metabolism , Microsomes, Liver/enzymology , Animals , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal/chemistry , Enzyme Inhibitors/administration & dosage , Enzyme Inhibitors/chemistry , Glucuronosyltransferase/antagonists & inhibitors , Glucuronosyltransferase/chemistry , Glucuronosyltransferase/genetics , Male , Mice , Microsomes, Liver/drug effects , RatsABSTRACT
To predit the mechanism of metabolic drug-drug interactions of hydroxygenkwanin with other drugs, we investigated the inhibition inhibitory effect of hydroxygenkwanin on UGTs and UGT1A1 activities of different liver microsomes. In the present study, 4-nitrophenol (4-NP) and ß-estradiol were elected as substrates to determine activities of UGTs and UGT1A1 by UV and HPLC, respectively. The results showed that, hydroxygenkwanin significantly inhibited UGTs activity in rat, mouse and human liver microsomes. UGT1A1 activity was inhibited by hydroxygenkwanin to varying degrees, with IC50 about 190, 10.93, 20.07, 76.31 µmolâ¢L⻹ in mouse liver microsome(MLM), rat liver microsome (RLM) and recombinant UGT1A1, and human liver microsome (HLM), respectively. The inhibition types were competitive inhibition (RLM, HLM) and linear mixed-typed linear inhibition (recombinant UGT1A1). The order for the inhibitory intensity was RLM>rUGT1A1>HLM>MLM. In conclusion, hydroxygenkwanin has an inhibitory effect on UGTs and UGT1A1 activities of different liver microsomes, with differences in species, indicating its potential drug interactions based on UGT1A1 enzyme. This study aims to provide a reliable experimental basis for its further research and development of hydroxygenkwanin, and provide theoretical reference for the clinic drug combination research.
Subject(s)
Enzyme Inhibitors/chemistry , Flavonoids/chemistry , Glucuronosyltransferase/antagonists & inhibitors , Animals , Chromatography, High Pressure Liquid , Glucuronosyltransferase/metabolism , Humans , Kinetics , Mice , Microsomes, Liver/chemistry , Microsomes, Liver/enzymologyABSTRACT
BACKGROUND AND PURPOSE: Small ubiquitin-like modifier (SUMO) conjugation is a post-translational modification associated with many human diseases. Characterization of the SUMO-modified proteome is pivotal to define the mechanistic link between SUMO conjugation and such diseases. This is particularly evident for SUMO2/3 conjugation, which is massively activated after brain ischemia/stroke, and is believed to be a protective response. The purpose of this study was to perform a comprehensive analysis of the SUMO3-modified proteome regulated by brain ischemia using a novel SUMO transgenic mouse. METHODS: To enable SUMO proteomics analysis in vivo, we generated transgenic mice conditionally expressing tagged SUMO1-3 paralogues. Transgenic mice were subjected to 10 minutes forebrain ischemia and 1 hour of reperfusion. SUMO3-conjugated proteins were enriched by anti-FLAG affinity purification and analyzed by liquid chromatography-tandem mass spectrometry. RESULTS: Characterization of SUMO transgenic mice demonstrated that all 3 tagged SUMO paralogues were functionally active, and expression of exogenous SUMOs did not modify the endogenous SUMOylation machinery. Proteomics analysis identified 112 putative SUMO3 substrates of which 91 candidates were more abundant in the ischemia group than the sham group. Data analysis revealed processes/pathways with putative neuroprotective functions, including glucocorticoid receptor signaling, RNA processing, and SUMOylation-dependent ubiquitin conjugation. CONCLUSIONS: The identified proteins/pathways modulated by SUMOylation could be the key to understand the mechanisms linking SUMOylation to neuroprotection, and thus provide new promising targets for therapeutic interventions. The new transgenic mouse will be an invaluable platform for analyzing the SUMO-modified proteome in models of human disorders and thereby help to mechanistically link SUMOylation to the pathological processes.
Subject(s)
Brain Ischemia/physiopathology , Ischemic Attack, Transient/physiopathology , Stroke/physiopathology , Ubiquitins/genetics , Ubiquitins/metabolism , Animals , Brain Ischemia/genetics , Brain Ischemia/metabolism , Ischemic Attack, Transient/genetics , Ischemic Attack, Transient/metabolism , Mass Spectrometry , Mice , Mice, Transgenic , Proteome/genetics , Proteome/metabolism , Proteomics , RNA Processing, Post-Transcriptional , SUMO-1 Protein/genetics , SUMO-1 Protein/metabolism , Small Ubiquitin-Related Modifier Proteins/genetics , Small Ubiquitin-Related Modifier Proteins/metabolism , Stroke/genetics , Stroke/metabolismABSTRACT
The Xenon flash lamp is a new type of light source for analytical instrument. The present paper analyzed the discharge process of xenon flash lamp, presented the discharge test system, and conducted experimental measurement of the voltage, current and optical pulse signal in the process of discharge. The results show that in the preliminary discharge, the free electron concentration was at a low level, so the energy was at a low level, then following the gas discharge, numerous free electrons formed in the lamp, resultin in the increase in the concentration of free electrons, therefore discharge current rised rapidly and voltage reduced. The lamp released photons to generate light pulse in the moment of ionic recombination, The pulse xenon lamp light energy output and spectral characteristic is related to electron energy in recombination and combination level of xenon, if the input energy and the energy consumption of the xenon lamp is inconsistent, it will lead to repeated capacitor charging and discharging and produce oscillation waveform. This paper is very useful for understanding the process of xenon lamp discharge, optimizing the driver circuit and the production of xenon flash lamp.