A carlactonoic acid methyltransferase that contributes to the inhibition of shoot branching in Arabidopsis.

SignificanceStrigolactones (SLs) are a group of apocarotenoid hormones, which regulates shoot branching and other diverse developmental processes in plants. The major bioactive form(s) of SLs as endogenous hormones has not yet been clarified. Here, we identify an Arabidopsis methyltransferase, CLAMT, responsible for the conversion of an inactive precursor to a biologically active SL that can interact with the SL receptor in vitro. Reverse genetic analysis showed that this enzyme plays an essential role in inhibiting shoot branching. This mutant also contributed to specifying the SL-related metabolites that could move from root to shoot in grafting experiments. Our work has identified a key enzyme necessary for the production of the bioactive form(s) of SLs.

Quantification and Characterization of Fine Plastic Particles as Considerable Components in Atmospheric Fine Particles.

The negative effects of air pollution, especially fine particulate matter (PM2.5, particles with an aerodynamic diameter of ≤2.5 µm), on human health, climate, and ecosystems are causing significant concern. Nevertheless, little is known about the contributions of emerging pollutants such as plastic particles to PM2.5 due to the lack of continuous measurements and characterization methods for atmospheric plastic particles. Here, we investigated the levels of fine plastic particles (FPPs) in PM2.5 collected in urban Shanghai at a 2 h resolution by using a novel versatile aerosol concentration enrichment system that concentrates ambient aerosols up to 10-fold. The FPPs were analyzed offline using the combination of spectroscopic and microscopic techniques that distinguished FPPs from other carbon-containing particles. The average FPP concentrations of 5.6 µg/m3 were observed, and the ratio of FPPs to PM2.5 was 13.2% in this study. The FPP sources were closely related to anthropogenic activities, which pose a potential threat to ecosystems and human health. Given the dramatic increase in plastic production over the past 70 years, this study calls for better quantification and control of FPP pollution in the atmosphere.

Oxycodone attenuates lipopolysaccharide-induced myocardial injury by inhibiting inflammation, oxidation and pyroptosis via Nrf2/HO-1 signalling pathway.

Myocardial injury and cardiovascular dysfunction are the most common complications of sepsis, and effective therapeutic candidate is still lacking. This study aims to investigate the protective effect of oxycodone in myocardial injury of lipopolysaccharide-induced sepsis and its related signalling pathways. Wild-type and nuclear factor erythroid 2-related factor 2 (Nrf2)-knockout mice, as well as H9c2 cardiomyocytes cultures treated with lipopolysaccharide (LPS) were used as models of septic myocardial injury. H9c2 cardiomyocytes culture showed that oxycodone protected cells from pyroptosis induced by LPS. Mice model confirmed that oxycodone pretreatment significantly attenuated myocardial pathological damage and improved cardiac function demonstrated by increased ejection fraction (EF) and fractional shortening (FS), as well as decreased cardiac troponin I (cTnI) and creatine kinase isoenzymes MB (CK-MB). Oxycodone also reduced the levels of inflammatory factors and oxidative stress damage induced by LPS, which involves pyroptosis-related proteins including: Nod-like receptor protein 3 (NLRP3), Caspase 1, Apoptosis-associated speck-like protein contain a CARD (ASC), and Gasdermin D (GSDMD). These changes were mediated by Nrf2 and heme oxygenase-1 (HO-1) because Nrf2-knockout mice or Nrf2 knockdown in H9c2 cells significantly reversed the beneficial effect of oxycodone on oxidative stress, inflammatory responses and NLRP3-mediated pyroptosis. Our findings yielded that oxycodone therapy reduces LPS-induced myocardial injury by suppressing NLRP3-mediated pyroptosis via the Nrf2/HO-1 signalling pathway in vivo and in vitro.

Clinical evaluation of photodynamic therapy for oral leukoplakia: a retrospective study of 50 patients.

BACKGROUND: Topical photodynamic therapy (PDT) has demonstrated encouraging results in the treatment of oral leukoplakia (OLK). However, data on the clinical efficacy of PDT in Chinese patients with OLK are still limited. METHODS: Fifty patients diagnosed with OLK were enrolled, including patients with various dysplastic tissues. All patients received topical PDT with 5-aminolevulinic acid (5-ALA) as a photosensitizer. Clinical efficacy was evaluated 4 weeks after treatment. Follow-up was performed every 3 months during the first year and every 6 months during the second year. RESULTS: The overall response rate was 68% (34/50): 12% (n = 6) complete and 56% (n = 28) partial responses. Aneuploidy was reduced in the patients with dysplastic lesions. Oral pain and local ulcers developed in 52% of the patients (n = 26). Patients with a long history of OLK including hyperplasia and dysplastic lesions, as well as those with non-homogenous lesions, were more likely to develop pain and ulcer. During follow-up, the recurrence rate of hyperplasia and dysplastic lesions was 32% (n = 16) and the malignant transformation rate of dysplastic lesions was 4% (n = 2). Lesions on the buccal mucosa were associated with recurrence (P = 0.044; OR: 0.108, 95% CI: 0.013-0.915). CONCLUSION: Topical 5-ALA-mediated PDT is an effective treatment for OLK, particularly for homogenous leukoplakia, with few side effects. The buccal mucosa may be a protective factor that can reduce recurrence.

Tomato geranylgeranyl diphosphate synthase isoform 1 is involved in the stress-triggered production of diterpenes in leaves and strigolactones in roots.

Carotenoids are photoprotectant pigments and precursors of hormones such as strigolactones (SL). Carotenoids are produced in plastids from geranylgeranyl diphosphate (GGPP), which is diverted to the carotenoid pathway by phytoene synthase (PSY). In tomato (Solanum lycopersicum), three genes encode plastid-targeted GGPP synthases (SlG1 to SlG3) and three genes encode PSY isoforms (PSY1 to PSY3). Here, we investigated the function of SlG1 by generating loss-of-function lines and combining their metabolic and physiological phenotyping with gene co-expression and co-immunoprecipitation analyses. Leaves and fruits of slg1 lines showed a wild-type phenotype in terms of carotenoid accumulation, photosynthesis, and development under normal growth conditions. In response to bacterial infection, however, slg1 leaves produced lower levels of defensive GGPP-derived diterpenoids. In roots, SlG1 was co-expressed with PSY3 and other genes involved in SL production, and slg1 lines grown under phosphate starvation exuded less SLs. However, slg1 plants did not display the branched shoot phenotype observed in other SL-defective mutants. At the protein level, SlG1 physically interacted with the root-specific PSY3 isoform but not with PSY1 and PSY2. Our results confirm specific roles for SlG1 in producing GGPP for defensive diterpenoids in leaves and carotenoid-derived SLs (in combination with PSY3) in roots.

Amide Proton Transfer Weighted MR Imaging for Predicting Meningioma Stiffness: A Feasibility Study.

BACKGROUND: Stiffness of meningioma is an important factor affecting the surgical resection and the prognosis of patients. PURPOSE: To examine the feasibility of APTw-magnetic resonance imaging (MRI) in evaluating meningioma stiffness. STUDY TYPE: Retrospective. POPULATION: Seventy-one patient with meningiomas, 39 were male and 32 were female; the mean age was 51 ± 10 years. FIELD STRENGTH/SEQUENCE: 3.0T; Turbo-spin-echo T1 -weighted and Gd-T1 -weighted sequence; Turbo-spin-echo T2 -weighted sequence; 2D fat-suppressed, turbo-spin-echo APTw pulse sequence. ASSESSMENT: The T1 WI signal intensity score, T2 WI signal intensity score, APTwmin , APTwmax , and APTwmean values were compared between soft, medium stiff and stiff meningiomas or non-stiff meningiomas and stiff meningiomas group. STATISTICAL TESTS: Chi-square test, one-way ANOVA analysis, independent-samples t-test, intra-class correlation coefficient, rank-sum test, receiver operating characteristic curve analysis. P < 0.05 was considered statistically significant in all tests. RESULTS: APTwmin and APTwmean in the stiff group were significantly lower than that in the non-stiff group (2.79% ± 0.42% vs. 1.90% ± 0.60% and 3.20% ± 0.31% vs. 2.55% ± 0.61%). APTwmin and APTwmean in the stiff group were significantly lower than that in the medium stiff and soft groups (1.90% ± 0.60% vs. 2.69% ± 0.40% and 3.12% ± 0.32%, 2.55% ± 0.61% vs. 3.17% ± 0.33% and 3.39% ± 0.18%), APTwmin in the medium stiff group was significantly lower than in the soft group, there was no significant difference in APTwmean between the medium stiff and soft groups (P = 0.190). APTwmin showed the best diagnostic performance for evaluating meningioma stiffness with an area under the curve of 0.913, when the APTwmin was lower than 2.4%, the meningioma was defined as a stiff tumor, the sensitivity, specificity, and accuracy were 87.1%, 87.5%, and 85.9%, respectively. DATA CONCLUSION: APTw-MRI could be used to evaluate meningioma stiffness, with APTwmin having the best evaluative efficiency. LEVEL OF EVIDENCE: 3 TECHNICAL EFFICACY: Stage 1.

Synthesis and Study of a Metal-Organic Framework-based Sulfite Fluorescence Sensor Modified with Urushiol.

Sulfites can pollute the environment and pose a great risk to human health in daily life, so there is an urgent need to develop efficient and lightweight sulfite detection materials. In this study, metal-organic framework-5-NH2/urushiol/PVP nanofiber composite films were prepared by an electrospinning technique for the fluorescence detection of sulfites. The results showed that the composite film could resist sulfuric acid corrosion at a concentration of 80% and inactivate Escherichia coli and Staphylococcus aureus at a concentration of 99%, and its maximum tensile strength was increased from the initial 2.753 to 4.145 N. The composite film was sensitive and specific for the fluorescence detection of sulfite.

Impact of New-Onset Conduction Disturbances following Transcatheter Aortic Valve Replacement on Outcomes: A Single-Center Study.

Background: Transcatheter aortic valve replacement (TAVR) is known to increase the incidence of conduction disturbances compared to surgical aortic valve replacement; however, there are limited data on the impact and duration of these conduction disturbances on longer term outcomes. Objective: To determine the differential impact of persistent versus nonpersistent new-onset conduction disturbances on TAVR-related complications and outcomes. Methods: This is a single-center retrospective analysis of 927 consecutive patients with aortic stenosis who underwent TAVR at Yale New Haven Hospital from July 2012 to August 2019. Patients with new-onset conduction disturbances within 7 days following TAVR were selected for this study. Persistent and nonpersistent disturbances were, respectively, defined as persisting or not persisting on all patient ECGs for up to 1.5 years after TAVR or until death. Results: Within 7 days after TAVR, conduction disturbances occurred in 42.3% (392/927) of the patients. Conduction disturbances persisted in 150 (38%) patients and did not persist in 187 (48%) patients, and 55 (14%) patients were excluded for having mixed (both persistent and nonpersistent) disturbances. Compared with nonpersistent disturbances, patients with persistent disturbances were more likely to receive a PPM within 7 days after the TAVR procedure (46.0% versus 4.3%, p < 0.001) and had a greater unadjusted 1-year cardiac-related and all-cause mortality risk (HR 2.54, p=0.044 and HR 1.90, p=0.046, respectively). Conclusion: Persistent conduction disturbances were associated with a greater cardiac and all-cause mortality rate at one year following TAVR. Future research should investigate periprocedural factors to reduce persistent conduction disturbances and outcomes beyond one year follow-up.

Phase coexistence in [C22/C1MIm]+[NO3]- ionic-liquid mixtures and first-order phase transitions from homogeneous liquid to smectic B by varying the cation ratio.

We perform molecular dynamics simulations to investigate the transition processes of [C22/C1MIm]+[NO3]- binary mixtures by varying the cation ratio of C22 to C1 at a fixed temperature of 400 K. The cation ratio is tuned by ranging C22 percentage from 0% to 100% with a fixed number of 4096 total simulated ion pairs. Our simulated-annealing results indicate that, at 400 K, pure C1 is a homogeneous liquid whilst pure C22 is an ionic liquid crystal (ILC) of smectic-B (SmB) type. With increasing C22 percentage, the system goes through a first-order phase transition from homogeneous liquid to nano-fragment liquid in the range from 15% to 17.5%, during which some of the individual cationic alkyl side chains locally aggregate to form small bundles "floating" in the polar "solvent" composed of anions and cationic head groups. Although the side chains in each bundle are parallelly aligned, the bundles distribute randomly without a global orientation. As the C22 percentage further increases, another first-order phase transition occurs to bring the system into the SmB ILC phase. Particularly, when the C22 percentage is in the range from 45% to 50%, the SmB phase coexists with the liquid phase containing both individual and bundled alkyl side chains.

The impacts of knowledge and attitude on behavior of antibiotic use for the common cold among the public and identifying the critical behavioral stage: based on an expanding KAP model.

BACKGROUND: This study aims to explore the impacts of knowledge and attitude on the behavior of antibiotic use during the treatment of the common cold based on the expanding KAP model, and then identify the critical behavioral stage. METHODS: A cross-sectional study was conducted on 815 public from 21 community health centers (CHCs) in Chongqing, China. Based on the expanding KAP model, a self-administered questionnaire was designed to measure knowledge, attitude, multi-stage behavior, and perceived threat, in which multi-stage behavior was divided into pre-use antibiotic behavior, during-use antibiotic behavior, and post-use antibiotic behavior. A structural equation model was used to examine the model fit and the direct, indirect, mediating effects, and moderating effect of the variables. RESULTS: The expanding KAP showed good model fit indices with χ²/df = 0.537, RMSEA = 0.033, CFI = 0.973, GFI = 0.971, NFI = 0.934, TLI = 0.979. Knowledge had a positive effect on attitude (ß = 0.503, p < 0.05), pre-use antibiotic behavior (ß = 0.348, p < 0.05), during-use antibiotic behavior (ß = 0.461, p < 0.001), and post-use antibiotic behavior (ß = 0.547, p < 0.001). Attitude had a positive effect on during-use antibiotic behavior (ß = 0.296, p < 0.001), and post-use antibiotic behavior (ß = 0.747, p < 0.001). The mediating effect of attitude was positive among knowledge, during-use antibiotic behavior (ß = 0.149, p < 0.05), and post-use antibiotic behavior (ß = 0.376, p < 0.001). Perceived threat also had a positive moderating effect between knowledge and post-use antibiotic behavior (ß = 0.021, p < 0.001). CONCLUSIONS: Knowledge, attitude and perceived threat had different effects on different stages of antibiotic behavior. The critical behavioral stage prioritized the post-use antibiotic behavior and during-use antibiotic behavior over pre-use antibiotic behavior.

Tetramethylpyrazine ameliorates endotoxin-induced acute lung injury by relieving Golgi stress via the Nrf2/HO-1 signaling pathway.

PURPOSE: Endotoxin-induced acute lung injury (ALI) is a severe disease caused by an imbalanced host response to infection. It is necessary to explore novel mechanisms for the treatment of endotoxin-induced ALI. In endotoxin-induced ALI, tetramethylpyrazine (TMP) provides protection through anti-inflammatory, anti-apoptosis, and anti-pyroptosis effects. However, the mechanism of action of TMP in endotoxin-induced ALI remains unclear. Here, we aimed to determine whether TMP can protect the lungs by inhibiting Golgi stress via the Nrf2/HO-1 pathway. METHODS AND RESULTS: Using lipopolysaccharide (LPS)-stimulated C57BL/6J mice and MLE12 alveolar epithelial cells, we observed that TMP pretreatment attenuated endotoxin-induced ALI. LPS + TMP group showed lesser lung pathological damage and a lower rate of apoptotic lung cells than LPS group. Moreover, LPS + TMP group also showed decreased levels of inflammatory factors and oxidative stress damage than LPS group (P < 0.05). Additionally, LPS + TMP group presented reduced Golgi stress by increasing the Golgi matrix protein 130 (GM130), Golgi apparatus Ca2+/Mn2+ ATPases (ATP2C1), and Golgin97 expression while decreasing the Golgi phosphoprotein 3 (GOLPH3) expression than LPS group (P < 0.05). Furthermore, TMP pretreatment promoted Nrf2 and HO-1 expression (P < 0.05). Nrf2-knockout mice or Nrf2 siRNA-transfected MLE12 cells were pretreated with TMP to explore how the Nrf2/HO-1 pathway affected TMP-mediated Golgi stress in endotoxin-induced ALI models. We observed that Nrf2 gene silencing partially reversed the alleviating effect of Golgi stress and the pulmonary protective effect of TMP. CONCLUSION: Our findings showed that TMP therapy reduced endotoxin-induced ALI by suppressing Golgi stress via the Nrf2/HO-1 signaling pathway in vivo and in vitro.

Accuracy of contrast-enhanced ultrasound in diagnosing extracranial carotid occlusion: A meta-analysis.

AIM: This study aimed to assess the accuracy of contrast-enhanced ultrasound (CEUS) in detecting extracranial carotid artery occlusion. MATERIALS AND METHODS: A systematic literature search was conducted in the Cochrane, PubMed, and EMBASE databases. Prospective or retrospective studies that reported sensitivity and specificity of CEUS for the diagnosis of carotid artery occlusion were selected. Eight studies (354 arteries) were included in the meta-analysis. A bivariate random-effect model was used to estimate overall sensitivity and specificity. The results were also summarized by developing a summary receiver operating characteristic (SROC) curve. RESULTS: The overall sensitivity, specificity, positive, and negative likelihood ratios were 0.99 (95% CI: 0.83-1.00), 0.97 (95% CI: 0.90-0.99), 30.0 (95% CI: 9.8-91.4), and 0.01 (95% CI: 0.00-0.21), respectively; the odds ratio for diagnosis was 4,796 (95% CI: 119-192,584). CONCLUSION: The diagnostic test accuracy suggests that CEUS is a reliable tool for diagnosis of extracranial carotid artery occlusion.

Is it really safe to replace decabromodiphenyl ether (BDE209) with decabromodiphenyl ethane (DBDPE)?: A perspective from hepatotoxicity.

In this paper, the hepatocytotoxicity and aryl hydrocarbon receptor (AHR) activity of decabromodiphenyl ethane (DBDPE), decabromodiphenyl ether (BDE209) and other 18 analogues were evaluated in vitro using human normal liver cell L02. These dioxin-like compounds showed differential hepatocytotoxicity (EC50  = 0.38-17.87 mg/L) and AHR activity (EROD activity = 4.53-46.35 U/µg). In silico study indicated the distance of π-π bonds between the benzene ring of compounds and residue Phe234 of AHR played a key role in the binding of AHR, and the substituents on the benzene ring also influenced the activity. Combining molecular biology and bioomics, the comprehensive investigations on the hepatotoxic mechanisms have demonstrated the AHR signaling pathway was the key mediation mechanism for the hepatotoxicity of DBDPE/BDE209. The cytochrome P450s (CYP2 family) mediated formation of reactive oxygenated intermediates might be the dominant toxic mechanism, which could produce oxidative stress or cause genotoxicity. Although the experimental toxicity of DBDPE was smaller relative to BDE209, the health risk of DBDPE may be much greater than we expected, due to the high potential to form a variety of dioxin-like intermediates by microbial oxidation of ethyl group. Therefore, whether it is really safe to replace BDE209 with DBDPE is a debatable question, and more ecotoxicological and health data are needed to clarify this issue.

Environment-Aware Rate Adaptation Based on Occasional Request and Robust Adjustment in 802.11 Networks.

The IEEE 802.11 standard provides multi-rate support for different versions. As there is no specification on the dynamic strategy to adjust the rate, different rate adaptation algorithms are applied according to different manufacturers. Therefore, it is often hard to interpret the performance discrepancy of various devices. Moreover, the ever-changing channels always challenge the rate adaptation, especially in the scenario with scarce spectrum and low SNR. As a result, it is important to sense the radio environment cognitively and reduce the unnecessary oscillation of the transmission rate. In this paper, we propose an environment-aware robust (EAR) algorithm. This algorithm employs an occasional small packet, designs a rate scheme adaptive to the environment, and enhances the robustness. We verify the throughput of EAR using network simulator NS-3 in terms of station number, motion speed and node distance. We also compare the proposed algorithm with three benchmark methods: AARF, RBAR and CHARM. Simulation results demonstrate that EAR outperforms other algorithms in several wireless environments, greatly improving the system robustness and throughput.

Adsorption Studies on the Removal of Anionic and Cationic Dyes from Aqueous Solutions Using Discarded Masks and Lignin.

The carbon materials derived from discarded masks and lignin are used as adsorbent to remove two types of reactive dyes present in textile wastewater: anionic and cationic. This paper introduces the results of batch experiments where Congo red (CR) and Malachite green (MG) are removed from wastewater onto the carbon material. The relationship between adsorption time, initial concentration, temperature and pH value of reactive dyes was investigated by batch experiments. It is discovered that pH 5.0-7.0 leads to the maximum effectiveness of CR and MG removal. The equilibrium adsorption capacities of CR and MG are found to be 232.02 and 352.11 mg/g, respectively. The adsorption processes of CR and MG are consistent with the Freundlich and Langmuir adsorption models, respectively. The thermodynamic processing of the adsorption data reveals the exothermic properties of the adsorption of both dyes. The results show that the dye uptake processes follow secondary kinetics. The primary adsorption mechanisms of MG and CR dyes on sulfonated discarded masks and alkaline lignin (DMAL) include pore filling, electrostatic attraction, π-π interactions and the synergistic interactions between the sulphate and the dyes. The synthesized DMAL with high adsorption efficiency is promising as an effective recyclable adsorbent for adsorbing dyes, especially MG dyes, from wastewater.

Full-Chain FeCl3 Catalyzation Is Sufficient to Boost Cellulase Secretion and Cellulosic Ethanol along with Valorized Supercapacitor and Biosorbent Using Desirable Corn Stalk.

Cellulosic ethanol is regarded as a perfect additive for petrol fuels for global carbon neutralization. As bioethanol conversion requires strong biomass pretreatment and overpriced enzymatic hydrolysis, it is increasingly considered in the exploration of biomass processes with fewer chemicals for cost-effective biofuels and value-added bioproducts. In this study, we performed optimal liquid-hot-water pretreatment (190 °C for 10 min) co-supplied with 4% FeCl3 to achieve the near-complete biomass enzymatic saccharification of desirable corn stalk for high bioethanol production, and all the enzyme-undigestible lignocellulose residues were then examined as active biosorbents for high Cd adsorption. Furthermore, by incubating Trichoderma reesei with the desired corn stalk co-supplied with 0.05% FeCl3 for the secretion of lignocellulose-degradation enzymes in vivo, we examined five secreted enzyme activities elevated by 1.3-3.0-fold in vitro, compared to the control without FeCl3 supplementation. After further supplying 1:2 (w/w) FeCl3 into the T. reesei-undigested lignocellulose residue for the thermal-carbonization process, we generated highly porous carbon with specific electroconductivity raised by 3-12-fold for the supercapacitor. Therefore, this work demonstrates that FeCl3 can act as a universal catalyst for the full-chain enhancement of biological, biochemical, and chemical conversions of lignocellulose substrates, providing a green-like strategy for low-cost biofuels and high-value bioproducts.

OsClo5 functions as a transcriptional co-repressor by interacting with OsDi19-5 to negatively affect salt stress tolerance in rice seedlings.

Caleosins constitute a small protein family with one calcium-binding EF-hand motif. They are involved in the regulation of development and response to abiotic stress in plants. Nevertheless, how they impact salt stress tolerance in rice is largely unknown. Thereby, biochemical and molecular genetic experiments were carried out, and the results revealed that OsClo5 was able to bind calcium and phospholipids in vitro and localized in the nucleus and endoplasmic reticulum in rice protoplasts. At the germination and early seedlings stages, overexpression transgenic lines and T-DNA mutant lines exhibited reduced and increased tolerance to salt stress, respectively, compared with the wild-type. Yeast two-hybrid, bimolecular fluorescence complementation and in vitro pull-down assays demonstrated that the EF-hand motif of OsClo5 was essential for the interactions with itself and OsDi19-5. Yeast one-hybrid, electrophoretic migration shift and dual-luciferase reporter assays identified OsDi19-5 as a transcriptional repressor via the TACART cis-element in the promoters of two salt stress-related target genes, OsUSP and OsMST. In addition, OsClo5 enhanced the inhibitory effect of OsDi19-5 in the tobacco transient system, which was confirmed by qRT-PCR analysis in rice seedlings under salt stress. The collective results deepen the understanding of the molecular mechanism underlying the roles of caleosin in the salt stress response. These findings will also inform efforts to improve salt tolerance of rice.

Positive interaction between GPER and ß-alanine in the dorsal root ganglion uncovers potential mechanisms: mediating continuous neuronal sensitization and neuroinflammation responses in neuropathic pain.

BACKGROUND: The pathogenesis of neuropathic pain and the reasons for the prolonged unhealing remain unknown. Increasing evidence suggests that sex oestrogen differences play a role in pain sensitivity, but few studies have focused on the oestrogen receptor which may be an important molecular component contributing to peripheral pain transduction. We aimed to investigate the impact of oestrogen receptors on the nociceptive neuronal response in the dorsal root ganglion (DRG) and spinal dorsal horn using a spared nerve injury (SNI) rat model of chronic pain. METHODS: We intrathecally (i.t.) administered a class of oestrogen receptor antagonists and agonists intrathecal (i.t.) administrated to male rats with SNI or normal rats to identify the main receptor. Moreover, we assessed genes identified through genomic metabolic analysis to determine the key metabolism point and elucidate potential mechanisms mediating continuous neuronal sensitization and neuroinflammatory responses in neuropathic pain. The excitability of DRG neurons was detected using the patch-clamp technique. Primary culture was used to extract microglia and DRG neurons, and siRNA transfection was used to silence receptor protein expression. Immunofluorescence, Western blotting, RT-PCR and behavioural testing were used to assess the expression, cellular distribution, and actions of the main receptor and its related signalling molecules. RESULTS: Increasing the expression and function of G protein-coupled oestrogen receptor (GPER), but not oestrogen receptor-α (ERα) and oestrogen receptor-ß (ERß), in the DRG neuron and microglia, but not the dorsal spinal cord, contributed to SNI-induced neuronal sensitization. Inhibiting GPER expression in the DRG alleviated SNI-induced pain behaviours and neuroinflammation by simultaneously downregulating iNOS, IL-1ß and IL-6 expression and restoring GABAα2 expression. Additionally, the positive interaction between GPER and ß-alanine and subsequent ß-alanine accumulation enhances pain sensation and promotes chronic pain development. CONCLUSION: GPER activation in the DRG induces a positive association between ß-alanine with iNOS, IL-1ß and IL-6 expression and represses GABAα2 involved in post-SNI neuropathic pain development. Blocking GPER and eliminating ß-alanine in the DRG neurons and microglia may prevent neuropathic pain development.

The tomato cytochrome P450 CYP712G1 catalyses the double oxidation of orobanchol en route to the rhizosphere signalling strigolactone, solanacol.

Strigolactones (SLs) are rhizosphere signalling molecules and phytohormones. The biosynthetic pathway of SLs in tomato has been partially elucidated, but the structural diversity in tomato SLs predicts that additional biosynthetic steps are required. Here, root RNA-seq data and co-expression analysis were used for SL biosynthetic gene discovery. This strategy resulted in a candidate gene list containing several cytochrome P450s. Heterologous expression in Nicotiana benthamiana and yeast showed that one of these, CYP712G1, can catalyse the double oxidation of orobanchol, resulting in the formation of three didehydro-orobanchol (DDH) isomers. Virus-induced gene silencing and heterologous expression in yeast showed that one of these DDH isomers is converted to solanacol, one of the most abundant SLs in tomato root exudate. Protein modelling and substrate docking analysis suggest that hydroxy-orbanchol is the likely intermediate in the conversion from orobanchol to the DDH isomers. Phylogenetic analysis demonstrated the occurrence of CYP712G1 homologues in the Eudicots only, which fits with the reports on DDH isomers in that clade. Protein modelling and orobanchol docking of the putative tobacco CYP712G1 homologue suggest that it can convert orobanchol to similar DDH isomers as tomato.

Adenosine improves LPS-induced ROS expression and increasing in monolayer permeability of endothelial cell via acting on A2AR.

Blood-labyrinth barrier (BLB) disruption plays a crucial role in the development of otitis media. The aims of our study was to explore the role and action mechanism of adenosine in LPS-induced endothelial cells (ECs) damage, which are one of the major principal cell type for blood-labyrinth barrier (BLB), and so as to assess the potential of adenosine to be used in the treatment of BLB disruption in animal experiment. In our study, ECs were treated with LPS to mimic BLB damage in vitro. Our data showed that adenosine at dosage of 1, 10, and 20 µM had no influence on the cell viability of ECs. LPS treatment obviously suppressed the expression of Occludin and Zonula occludens-1 (ZO-1) in ECs, which was partly recused by adenosine treatment. Meantime, LPS-induced increasing in reactive oxygen species (ROS) production and ECs permeability also was rescued by adenosine treatment. However, inhibition the A2A receptor (A2AR) could attenuate the influence of adenosine on LPS-treated ECs, indicating that adenosine alleviated LPS-induced BLB damage by activating A2AR. Moreover, the inhibition of adenosine to LPS-induced inactivation of AMPK/AKT signaling pathway was partly recused by A2AR suppression. In addition, Compound C (an AMPK inhibitor) decreased the expression of Occludin and ZO-1 in ECs following LPS combined with adenosine treatment. In conclusion, adenosine alleviates LPS-induced BLB damage via AMPK/AKT pathway through activation of A2AR. This work suggests that adenosine may be a candidate drug for the treatment of BLB dysfunction-related diseases.