BF3 complexing phosphate/phosphonate ionic liquids: synthesis, characterization and thermophysical properties.

A new group of BF3 complexing phosphate/phosphonate ionic liquids (ILs) [Emim][X(BF3)2] (X = dimethyl phosphate, diethyl phosphate, methyl phosphonate, and ethyl phosphonate) were synthesized and characterized. Key thermophysical properties of the new complex ionic liquids, including density, viscosity, conductivity, surface tension, solid-liquid phase transition, and thermal stability were determined and compared with those of [Emim][X]. Some other important thermophysical properties such as isobaric thermal expansion coefficient, molecular volume, standard molar entropy, and lattice potential energy were obtained from measured density data, and the free volume was estimated by a linear equation presented in this article, while critical temperature, normal boiling temperature, and enthalpy of vaporization were estimated from measured surface tension and density data. Furthermore, Fragility study shows that [Emim][X(BF3)2] could be considered as metallic glass-forming liquids, while [Emim][X] could be considered as extremely fragile liquids. The ionicity of [Emim][X(BF3)2] was predicted by Walden rule, and the result shows that these ILs fit well with Walden law. The key features of these complex ILs are their extremely low glass transition (-95.33~-98.46 ℃) without melting, considerably low viscosities (33.876~58.117 mPa·s), and high values of free volume fraction (comparable to [Omim][BF4], [Emim][NTf2], and [Emim][TCB]).

Phytochemical composition of Tibetan tea fermented by Eurotium cristatum and its effects on type 1 diabetes mice and gut microbiota.

"Golden-flower" Tibetan tea (GTT) is an innovative dark tea fermented via fungus Eurotium cristatum. To study GTT effects on alleviating the symptoms of type 1 diabetes mellitus (T1DM), GTT's extract (GTTE) was prepared. GTTE chemical compositions were analyzed via HPLC, pyrolysis-gas chromatography-mass (Py-GC-MS) spectrometry analysis, and chemistry analyses. GTTE effects on T1DM were explored on T1DM mice model induced by streptozotocin (STZ). GTTE was composed mainly of tea pigment theabrownin (TB) (49.18%), with high percentages of polysaccharide (16.93%), protein (10.15%), polyphenols (13.90%), amino acids (5.89%), caffeine (1.83%), and flavonoids (0.67%). Py-GC-MS results exhibited that GTTE constituted of phenols, lipids, sugars, and proteins. GTTE attenuated T1DM conditions of mice, relieved their liver and pancreatic injury, restored damaged islet cells, decreased oxidative stress by increasing superoxide dismutase (SOD) and catalase (CAT) levels, modulated cytokine expression leading to the decreasing pro-inflammatory cytokines TNF-α and IL-6, increased anti-inflammatory cytokines IL-4 to improve inflammatory responses, and optimized gut microbiota composition and structure based on high-throughput 16S rDNA sequencing, suggesting multi-channel anti-diabetes mechanisms.

Synthesis, Characterization, and Physicochemical Properties of New [Emim][BF3X] Complex Anion Ionic Liquids.

A new series of complex anion ionic liquids (ILs) [Emim][BF3X] (X = CH3SO3, EtSO4, HSO4, Tosylate) were synthesized and characterized by nuclear magnetic resonance, elemental analysis, differential scanning calorimetry analysis, and thermogravimetry. The physicochemical properties of these ILs, such as density, viscosity, conductivity, and surface tension, were measured and correlated with thermodynamic and empirical equations in the temperature range of 293.15-358.15 K under ambient conditions, and the thermal expansion coefficient, standard molar entropy, lattice potential energy, viscosity activation energy, surface enthalpy, and surface entropy were further calculated from experimental values. According to the temperature-dependent viscosity and conductivity, [Emim][BF3X] ILs follow the Walden rule, and they are classified as "good (or super) ionic liquids".

MicroRNA-15a/ß1,4-GalT-I axis contributes to cartilage degeneration via NF-κB signaling in osteoarthritis.

OBJECTIVE: Osteoarthritis is a condition characterized by articular cartilage degradation. The increased expression of ß1,4-Galactosyltransferase-I (ß1,4-GalT-I) in the articular cartilage of osteoarthritis patients was related to an inflammatory response. The aim of this study was to elucidate the role of ß1,4-GalT-I in osteoarthritis. This study aimed to determine the function of 1,4-GalT-I in osteoarthritis. METHODS: The osteoarthritis mouse model with the destabilization of the medial meniscus was established by microsurgical technique. Pathological changes in articular cartilage were observed by hematoxylin and eosin staining and safranin O-fast green staining. Quantitative real-time polymerase chain reaction, western blot, and enzyme-linked immunosorbent assays were used to observe mRNA and protein expression, respectively. RNA interactions were verified by a luciferase reporter assay. SA-ß-Gal staining was used to assess chondrocyte senescence. Immunofluorescence staining was conducted to observe the localization of Nuclear Factor-kappaB (NF-κB). RESULTS: ß1,4-GalT-I and microRNA-15a (miR-15a) show high and low expression in the articular cartilage of osteoarthritis, respectively. MiR-15a inhibits the mRNA translation of ß1,4-GalT-I. ß1,4-GalT-I promotes extracellular matrix degradation, senescence, and NF-κB activation in IL-1ß-stimulated chondrocytes, which can be reversed by overexpression of miR-15a. Intra-articular injection of microRNA-15a ameliorates cartilage degeneration by inhibiting ß1,4-GalT-I and phosphorylation of NF-κB in vivo. CONCLUSION: The authors clarified that the miR-15a/ß1,4-GalT-I axis inhibits the phosphorylation of NF-κB thereby inhibiting extracellular matrix degradation and senescence in chondrocytes to alleviate cartilage degeneration in osteoarthritis. MiR-15a and ß1,4-GalT-I may serve as potentially effective targets for the future treatment of osteoarthritis.

[Effectiveness of arthroscopic binding fixation using suture through single bone tunnel for posterior cruciate ligament tibial insertion fracture in adults].

Objective: To explore the effectiveness of arthroscopic binding fixation using suture through single bone tunnel for posterior cruciate ligament (PCL) tibial insertion fractures in adults. Methods: Between October 2019 and October 2021, 16 patients with PCL tibial insertion fractures were treated with arthroscopic binding fixation using suture through single bone tunnel. There were 11 males and 5 females with an average age of 41.1 years (range, 26-58 years). The fractures were caused by traffic accident in 12 cases and sports in 4 cases. The time from injury to operation ranged from 2 to 10 days with an average of 6.0 days. The fractures were classified as Meyers-McKeever type Ⅱ in 4 cases and type Ⅲ in 9 cases, and Zaricznyi type Ⅳ in 3 cases. There were 2 cases of grade Ⅰ, 7 cases of grade Ⅱ, and 7 cases of grade Ⅲ in the posterior drawer test. There were 3 cases combined with lateral collateral ligament injury and 2 cases with meniscus injury. The visual analogue scale (VAS) score, Lysholm score, International Knee Documentation Committee (IKDC) score, and knee range of motion were used to evaluate knee joint function. The posterior drawer test and knee stability tester (Kneelax 3) were used to evaluate knee joint stability. The X-ray films were used to evaluate fracture reduction and healing. Results: All incisions healed by first intention after operation. There was no incision infection, popliteal neurovascular injury, or deep venous thrombosis of lower limbs. All patients were followed up 6-12 months, with an average of 10 months. X-ray films at 6 months after operation showed the fractures obtained bone union. There were 11 cases of grade 0, 4 cases of gradeⅠ, and 1 case of grade Ⅱin posterior drawer test, showing significant difference when compared with preoperative results ( Z=23.167, P<0.001). The VAS score, Lysholm score, IKDC score, knee range of motion, and the results of Kneelax3 examination all significantly improved when compared with preoperative results ( P<0.05). Conclusion: For adult patients with PCL tibial insertion fractures, the arthroscopic binding fixation using suture through single bone tunnel has the advantages of minimal trauma, good fracture reduction, reliable fixation, and fewer complications. The patient's knee joint function recovers well.

MicroRNA-15a/β1,4-GalT-I axis contributes to cartilage degeneration via NF-κB signaling in osteoarthritis

Abstract Objective Osteoarthritis is a condition characterized by articular cartilage degradation. The increased expression of β1,4-Galactosyltransferase-I (β1,4-GalT-I) in the articular cartilage of osteoarthritis patients was related to an inflammatory response. The aim of this study was to elucidate the role of β1,4-GalT-I in osteoarthritis. This study aimed to determine the function of 1,4-GalT-I in osteoarthritis. Methods The osteoarthritis mouse model with the destabilization of the medial meniscus was established by microsurgical technique. Pathological changes in articular cartilage were observed by hematoxylin and eosin staining and safranin O-fast green staining. Quantitative real-time polymerase chain reaction, western blot, and enzyme-linked immunosorbent assays were used to observe mRNA and protein expression, respectively. RNA interactions were verified by a luciferase reporter assay. SA-β-Gal staining was used to assess chondrocyte senescence. Immunofluorescence staining was conducted to observe the localization of Nuclear Factor-kappaB (NF-κB). Results β1,4-GalT-I and microRNA-15a (miR-15a) show high and low expression in the articular cartilage of osteoarthritis, respectively. MiR-15a inhibits the mRNA translation of β1,4-GalT-I. β1,4-GalT-I promotes extracellular matrix degradation, senescence, and NF-κB activation in IL-1β-stimulated chondrocytes, which can be reversed by overexpression of miR-15a. Intra-articular injection of microRNA-15a ameliorates cartilage degeneration by inhibiting β1,4-GalT-I and phosphorylation of NF-κB in vivo. Conclusion The authors clarified that the miR-15a/β1,4-GalT-I axis inhibits the phosphorylation of NF-κB thereby inhibiting extracellular matrix degradation and senescence in chondrocytes to alleviate cartilage degeneration in osteoarthritis. MiR-15a and β1,4-GalT-I may serve as potentially effective targets for the future treatment of osteoarthritis.

miR-653-5p suppresses the viability and migration of fibroblast-like synoviocytes by targeting FGF2 and inactivation of the Wnt/beta-catenin pathway.

BACKGROUND: Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease. Several studies reported that fibroblast-like synoviocytes (FLSs) and miRNAs are associated with RA pathogenesis. This study explored the function of miR-653-5p in the regulation of human fibroblast-like synoviocytes-rheumatoid arthritis (HFLS-RA) cells. METHODS: The mRNA and protein levels of genes were measured by RT-qPCR and western blot, respectively. MTT, wound healing, and invasion assays were used to evaluate the viability and metastasis of FLSs. Luciferase reporter and RNA pull-down assays were employed to determine the interaction between miR-653-5p and FGF2. RESULTS: RT-qPCR results demonstrated that miR-653-5p expression was decreased and FGF2 level was increased in synovial tissues and FLSs of RA. Moreover, the viability and metastasis of FLSs were accelerated by miR-653-5p addition, which was restrained by miR-653-5p suppression. Furthermore, we demonstrated that levels of Rac1, Cdc42, and RhoA were decreased after miR-653-5p addition. Besides, luciferase reporter and RNA pull-down assays implied that miR-653-5p targeted the 3'-UTR of FGF2. Functional assays showed that FGF2 overexpression neutralized the suppressive effects of miR-653-5p addition on HFLS-RA cell viability, metastasis, and the levels of Rho family proteins. Meanwhile, the levels of ß-catenin, cyclin D1, and c-myc were declined by miR-653-5p supplementation, but enhanced by FGF2 addition. CONCLUSION: In sum, we manifested that miR-653-5p restrained HFLS-RA cell viability and metastasis via targeting FGF2 and repressing the Wnt/beta-Catenin pathway.

Analysis of Heavy Metal Pollution in Cultivated Land of Different Quality Grades in Yangtze River Delta of China.

The distribution of heavy metal pollution in cultivated land is closely related to the quality of the cultivated land. In this study, 533 soil samples were collected from cultivated land in the Yangtze River delta region in China for Cd, Pb, and Hg analyses. Spatial statistical analysis was used to study the heavy metal pollution in the cultivated land, and the driving forces of heavy metal distribution in different cultivated land quality subdivisions were analyzed with GeogDetector. The conclusions are as follows: (1) Among the three heavy metals in the study area, the coefficient of variation of Cd is the largest, and that of Pb is the smallest. The proportion of Cd and Hg exceeding the standard value (the standard of level two in GB 15618-2018) is relatively large, both of which are 5%; (2) From the perspective of the spatial distribution of soil heavy metal pollution, only four counties (CX, HN, WY, and LH) were free of heavy metal pollution. Soil heavy metal pollution in AJ, SY, QJ, and DS counties is relatively serious, and the pollution may come from agricultural activities, manufacturing, and prevalent coastal shipping industries in these counties; (3) The heavy metal pollution levels of cultivated land with different quality levels are different. The high-quality cultivated land has no high contamination, while the medium and the general cultivated land both have high contamination. High contamination is related to Cd for medium and general cultivated lands, and to Hg in only general cultivated land; (4) The main driving factors of heavy metal concentration in cultivated soil were GDP, followed by soil organic matter, and pH. These results indicate that the spatial distribution of heavy metal concentration in cultivated soil was affected by the level of economic development, followed by the ecological environment, indicating that human activities had a critical impact on the ecological environment of cultivated land.

Reconstruction of circRNA-miRNA-mRNA associated ceRNA networks reveal functional circRNAs in intracerebral hemorrhage.

Circular RNA (circRNA), a novel class of noncoding RNAs, has been used extensively to complement transcriptome remodeling in the central nervous system, although the genomic coverage provided has rarely been studied in intracerebral hemorrhage (ICH) and is limited and fails to provide a detailed picture of the cerebral transcriptome landscape. Here, we described sequencing-based transcriptome profiling, providing comprehensive analysis of cerebral circRNA, messenger RNA (mRNA) and microRNA (miRNA) expression in ICH rats. In the study, male Sprague-Dawley rats were subjected to ICH, and next-generation sequencing of RNAs isolated from non-hemorrhagic (Sham) and hemorrhagic (ICH) rat brain samples collected 7 (early phase) and 28 (chronic phase) days after insults, was conducted. Bioinformatics analysis was performed to determine miRNA binding sites and gene ontology of circRNAs, target genes of miRNAs, as well as biological functions of mRNAs, altered after ICH. These analyses revealed different expression profiles of circRNAs, mRNAs and miRNAs in day-7 and day-28 ICH groups, respectively, compared with the Sham. In addition, the expression signature of circRNAs was more sensitive to disease progression than that of mRNAs or miRNAs. Further analysis suggested two temporally specific circRNA-miRNA-mRNA networks based on the competitive endogenous RNA theory, which had profound impacts on brain activities after ICH. In summary, these results suggested an important role for circRNAs in the pathogenesis of ICH and in reverse remodeling based on self-protection support, providing deep insights into diverse possibilities for ICH therapy through targeting circRNAs.

Research on automatic labeling of imbalanced texts of customer complaints based on text enhancement and layer-by-layer semantic matching.

Due to its potential impact on business efficiency, automated customer complaint labeling and classification are of great importance for management decision making and business applications. The majority of the current research on automated labeling uses large and well-balanced datasets. However, customer complaint labels are hierarchical in structure, with many labels at the lowest hierarchy level. Relying on lower-level labels leads to small and imbalanced samples, thus rendering the current automatic labeling practices inapplicable to customer complaints. This article proposes an automatic labeling model incorporating the BERT and word2vec methods. The model is validated on electric utility customer complaint data. Within the model, the BERT method serves to obtain shallow text tags. Furthermore, text enhancement is used to mitigate the problem of imbalanced samples that emerge when the number of labels is large. Finally, the word2vec model is utilized for deep text analysis. Experiments demonstrate the proposed model's efficiency in automating customer complaint labeling. Consequently, the proposed model supports enterprises in improving their service quality while simultaneously reducing labor costs.

Metal-free catalytic hydrocarboxylation of hexafluorobut-2-yne.

An efficient method for stereoselective synthesis of trifluorinated enol esters catalyzed by base was introduced. The DFT calculations and experimental results both supported the nucleophilic addition process. The protocol featured mild reaction conditions and showed a wide functional group tolerance. The one-pot simultaneous etherification and esterification of the salicylic acids further demonstrated the prospective synthetic application.

New insights into the fractionation of effluent organic matter on diagnosis of key composition affecting advanced phosphate removal by Zr-based nanocomposite.

The influence of effluent organic matter (EfOM) on phosphate removal by adsorption plays an important role in evaluating the applicability of adsorbents. Currently, molecular understanding of EfOM regarding its impact on adsorption is insufficient due to a lack of appropriate EfOM fractionation/characterization protocols, as associated with the specific structure-function property of adsorbents. In this work, a combined method coupling DEAE/XAD fractionation with molecular characterization was proposed, targeting the versatile structure-function characters of nanocomposite, to reveal the composition of EfOM as well as its impact on phosphate removal by nanocomposite during long-term adsorption/regeneration runs. Zirconium-based polystyrene anion exchanger (HZO-201) was selected as a representative nanocomposite, featuring with porous networking matrix, positively charged surface and multiple adsorptive sites. The EfOM samples from three biologically treated sewage effluent sources were separated into fractions of negatively charged organic acid (OA) and hydrophobic-, transphilic-, hydrophilic-neutral/base (HPO-n/b, TPI-n/b and HPI-n/b). The combined method enables effective differentiation of the charge, aromaticity, molecular weight and functionalities of the fractions, matching the multiple structural/surface characteristics of HZO-201 and favoring the evaluation on the impact of the EfOM fractions. The interference sequence of the EfOM fractions on phosphate removal followed an order of OA > HPO-n/b > TPI-n/b > HPI-n/b. The OA fraction, characterized by negatively charged, aromatic functionalities and broad molecular weight distribution (1-5 kDa and 14 kDa), was recognized as the key interfering fraction, presumably due to its multiple adsorption pathways (i.e., ion exchange, π-π interactions and pore filling). Particularly, the low-molecular-weight OA moieties (1-4 kDa) progressively accumulated onto the nanocomposite via irreversible adsorption, causing a continuous phosphate-capacity loss by 32.70% over multiple cycles. We believe the combined fractionation/characterization method may be extended to other complex water systems to identify key influential organic matters in polishing treatment of various pollutants by adsorption.

Elucidation of colon-protective efficacy of diosgenin in experimental TNBS-induced colitis: inhibition of NF-κB/IkB-α and Bax/Caspase-1 signaling pathways.

The aim of present investigation was to elucidate the unrevealed beneficial role of diosgenin against an experimental model of TNBS (2,4,6-trinitrobenzenesufonic acid)-induced ulcerative colitis (UC). Colitis was induced in Sprague-Dawley rats by intrarectal administration of TNBS (in 50% ethanol). Then animals were treated with diosgenin (50, 100, and 200 mg/kg) for 14 days. Various biochemical, behavioral, molecular, and histological analysis was performed. Diosgenin significantly decreased (p < 0.05) TNBS-induced elevated colonic oxido-nitrosative damage, myeloperoxidase, hydroxyproline, mRNA expressions of proinflammatory cytokines (TNF-α, IL-1ß, IL-6, and IFN-γ) and inflammatory markers (iNOs and COX-2) induced by TNBS. Western blot analysis relevated that TNBS-induced up-regulated protein expressions of NF-κB, IκBα, Bax, and Caspase-1 were markedly decreased (p < 0.05) by diosgenin treatment. It also markedly ameliorated the histological insults induced in the colon by TNBS. In conclusion, diosgenin exerts its colon-protective efficacy probably through the inhibition of NF-κB/IkB-α and Bax/Caspase-1 signaling pathways to experimental TNBS-induced ulcerative colitis. ABBREVIATIONS: ANOVA: Analysis of variance; 5-ASA: 5-aminosalicylic acid; Bax: Bcl-2-associated X protein; COX-2: Cyclooxygenase-2; DAI: Disease Activity Index; DMSO: Dimethyl sulfoxide; GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; GSH: Glutathione; HP: Hydroxyproline; IAEC: International Animal Ethics Committee; IBD: Inflammatory Bowel Disease; IBS: Inflammatory Bowel Syndrome; IL's: Interleukin's; IFN-γ: Interferon-gamma; IκBα: nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor-alpha; iNOs: Inducible nitric oxide synthase; LTB4: Leukotriene B4; MDA: Malondialdehyde; MPO: Myeloperoxidase; NO: Nitric Oxide; NF-κB: Nuclear Factor-κB; ROS: Reactive Oxygen Species; SOD: Superoxide Dismutase; TNBS: Trinitrobenzene Sulfonic Acid; TNF-α: Tumor necrosis factor-α.

Overexpression of SLFN5 induced the epithelial-mesenchymal transition in human lung cancer cell line A549 through ß-catenin/Snail/E-cadherin pathway.

Lung cancer is a disease with increasing morbidity worldwide in recent years. Approaches such as chemotherapy and biological targeting for its treatment are urgently needed. Epithelial-mesenchymal transition (EMT) is an important initiation stage for tumor cells to acquire invasive and metastatic abilities. Increasing findings have shown that human schlafen family member 5 (SLFN5) plays a key role in malignant tumors. However, the role of SLFN5 in lung cancer cells is not completely elucidated yet. In this study, overexpression or knockdown of SLFN5 gene were induced by lentiviral transfection in human lung cancer cell line A549, then the EMT of A549 was detected by green fluorescent protein labeling method, the migrative and invasive abilities were evaluated via transwell and wound-healing tests in vitro and chick chorioallantoic membrane inoculation in vivo, and the possible mechanism was studied by quantitative real-time PCR and Western blotting. Our results demonstrated that overexpression of SLFN5 promoted the morphology transformation of A549 from epithelial to mesenchymal, as well as migration and invasion. However, knockdown of SLFN5 resulted in the opposite results. Moreover, with the development of EMT after SLFN5 was overexpressed, A549 exhibited enhanced translocation of ß-catenin from membrane to cytoplasm or nucleus, with higher level of EMT-related transcription factor Snail, and lower expression of adhesin E-cadherin. Together these results suggest that SLFN5 may act as a synergist in lung cancer cell tumorigenesis and progression, providing a potential target for developing drugs for lung cancer therapy in future.

Lithium chloride promoted hematoma resolution after intracerebral hemorrhage through GSK-3ß-mediated pathways-dependent microglia phagocytosis and M2-phenotype differentiation, angiogenesis and neurogenesis in a rat model.

Some neuroprotective agents have been used clinically to address the resulting various adverse effects after intracerebral hemorrhage (ICH). Particularly, effectively removing the hematoma is of practical significance to exert neuroprotective effects following ICH. However, such agents are still in need of development. Lithium chloride (LiCl) has shown neuroprotective effects through glycogen synthase kinase-3ß (GSK-3ß) inhibition in a variety of central nervous system diseases. However, the impact of LiCl on hematoma clearance and the potential molecular mechanisms have not been reported. We hypothesize that LiCl may exert neuroprotective roles after ICH, partly through promoting hematoma resolution. In this study, male Sprague-Dawley rats were subjected to ICH followed by intraperitoneal injection of LiCl (60 mg/kg). The hematoma volumes of ipsilateral hemisphere were determined using Drabkin's method. The sensorimotor deficits were evaluated by neurobehavioral tests. The expressions of target molecules involved in the process of hematoma clearance were assayed using immunofluorescence and Western blot. Our results showed that animals treated with LiCl presented significantly reduced hematoma volume after ICH, which was coupled with enhanced microglia phagocytosis and its differentiation into M2-phenotype within the first 7 days and up-regulated angiogenesis and neurogenesis in the next 7 days. Meanwhile, GSK-3ß was inhibited by LiCl and ß-catenin became stabilized, which was followed by up-regulation of nuclear factor erythroid 2-related factor 2 and CD36 from days 3 to 7, and increase of vascular endothelial growth factor and brain-derived neurotrophic factor from days 7 to 14. These data suggest that LiCl promotes hematoma resolution via enhancing microglia phagocytosis and M2-phenotype differentiation in the early stage (< 7 days) and angiogenesis and neurogenesis in the chronic phase (days 7-14), thus eventually improving the functional outcomes of ICH rats.

Bipartite anchoring of SCREAM enforces stomatal initiation by coupling MAP kinases to SPEECHLESS.

Cell fate in eukaryotes is controlled by mitogen-activated protein kinases (MAPKs) that translate external cues into cellular responses. In plants, two MAPKs-MPK3 and MPK6-regulate diverse processes of development, environmental response and immunity. However, the mechanism that bridges these shared signalling components with a specific target remains unresolved. Focusing on the development of stomata-epidermal valves that are essential for gas exchange and transpiration-here, we report that the basic helix-loop-helix protein SCREAM functions as a scaffold that recruits MPK3/6 to downregulate SPEECHLESS, a transcription factor that initiates stomatal cell lineages. SCREAM directly binds to MPK3/6 through an evolutionarily conserved, yet unconventional, bipartite motif. Mutations in this motif abrogate association, phosphorylation and degradation of SCREAM, unmask hidden non-redundancies between MPK3 and MPK6, and result in uncontrolled stomatal differentiation. Structural analyses of MPK6 with a resolution of 2.75 Å showed bipartite binding of SCREAM to MPK6 that is distinct from an upstream MAPKK. Our findings elucidate, at the atomic resolution, the mechanism that directly links extrinsic signals to transcriptional reprogramming during the establishment of stomatal cell fate, and highlight a unique substrate-binding mode adopted by plant MAPKs.

BIO alleviates inflammation through inhibition of GSK-3ß in a rat model of intracerebral hemorrhage.

OBJECTIVE: Inflammation plays a key role in secondary brain damage following intracerebral hemorrhage (ICH). Glycogen synthase kinase-3ß (GSK-3ß) plays a strong proinflammatory role in many CNS diseases, including stroke. The present study was undertaken to examine the effects of 6-bromoindirubin-3'-oxime (BIO), a specific inhibitor of GSK-3ß, on inflammation in ICH rats. METHODS: An ICH rat model was induced by autologous whole-blood injection into the striatum. First, 10, 20, 40, 60, 80, or 100 µg/kg BIO was applied to ICH animals to determine an optimal dosage for producing sufficient GSK-3ß inhibition in rat ipsilateral hippocampus by Western blotting. Second, 40 µg/kg BIO was applied to ICH rats for 1, 3, 7, or 14 days, respectively, to determine a suitable intervention time course of BIO by Western blotting analysis on GSK-3ß. Third, Western blotting and enzyme-linked immunosorbent assay were used for quantification of inflammation-related factors upstream or downstream of GSK-3ß in rat ipsilateral hippocampus. Then, immunohistochemical staining was applied to detect activated microglia and apoptotic cells in rat ipsilateral hippocampus. Last, neurobehavioral tests were performed to assess the sensorimotor impairments in the ICH rats. RESULTS: The results show that BIO 1) blocked GSK-3ßTyr216 phosphorylation/activation, thus stabilizing ß-catenin, increasing upstream brain-derived neurotrophic factor and downstream heat shock protein 70 levels, and decreasing the levels of nuclear factor-κB p65 and cyclooxygenase 2; 2) decreased the levels of the proinflammatory cytokines tumor necrosis factor-α and interleukin (IL)-1ß and IL-6 and elevated the level of antiinflammatory cytokine IL-10; 3) inhibited microglia activation and cell apoptosis; and 4) improved the sensorimotor deficits of ICH rats. CONCLUSIONS: BIO posttreatment inhibited microglia activation, prevented inflammation and hippocampal cell death, and ameliorated functional and morphological outcomes in a rat ICH model through inactivation of GSK-3ß.

The neuroprotective effect of lithium chloride on cognitive impairment through glycogen synthase kinase-3ß inhibition in intracerebral hemorrhage rats.

To the clinical cognitive impairment following intracerebral hemorrhage, comprehensive neuropsychological assessments and efficacious interventions have rarely been conducted. Lithium chloride, a classical treatment for bipolar disorder, has shown neuroprotective effects through glycogen synthase kinase-3ß inhibition in a variety of central nervous system diseases, including stroke. Since neurons that contain glutamate play crucial roles in psychological functions, such as learning and memory, the glutamate-mediated excitotoxicity and consequent neuronal death and cognitive impairment in hippocampus may co-determine the clinical course of intracerebral hemorrhage. However, the potential molecular mechanisms have rarely been demonstrated in intracerebral hemorrhage researches. In this study, Male Sprague-Dawley rats, subjected to intrastriatal blood infusion, were treated with lithium chloride and underwent neurobehavioral test for equivalent injury severity and neurological functional deficits, Morris water maze test for cognitive impairment, high performance liquid chromatography analysis for excitotoxic index determination, immunohistochemistry analysis for neuronal apoptosis, and Western blot analysis for glycogen synthase kinase-3ß activity. Our results showed lithium chloride inhibited glycogen synthase kinase-3ß activation, which on one hand, suppressed downstream CRMP-2/NR2B, thus diminishing the excitotoxic index level; and on the other, stabilized ß-catenin, thus modulating its downstream apoptosis-related factors such as NF-κB, Bcl-2 and Bax. Meanwhile, glycogen synthase kinase-3ß inactivation was paralleled by decreased neuronal death, improved neurological functional deficits and ameliorated cognitive deficits in intracerebral hemorrhage animals. These findings indicate that lithium chloride improves glutamate-mediated excitotoxicity-induced cognitive deficits after intracerebral hemorrhage and that lithium chloride might be a potential therapeutic agent for brain damages caused by intracerebral hemorrhage.

Comparison of the Efficacy of Different Analgesia Treatments for Total Knee Arthroplasty: A Network Meta-Analysis.

BACKGROUND AND AIM: The severe pain after total knee arthroplasty (TKA) brings many patients more suffering, longer hospital stay, and higher expenses. This study was designed to assess the relative efficacy of several clinical treatments for postoperative analgesia of TKA through network meta-analysis based on multiple published randomized controlled trials. METHODS: Embase and PubMed were utilized to conduct this network meta-analysis from inception until 2016. Pain score, morphine consumption (milligrams), and length of hospitalization (day) were selected as the endpoints. RESULTS: A total of 58 studies with 3501 patients were included in this network meta-analysis. Except for patient-controlled epidural analgesia+femoral nerve block (FNB) and sciatic nerve block, all treatments were significantly superior to placebo in pain score 6 to 8 hours. In terms of pain score 24 hours, only continuous femoral nerve block (cFNB), periarticular infiltration, periarticular infiltration+FNB, single-dose FNB, and sciatic nerve block+FNB exhibited better performance than control group. For pain score 48 hours after surgery, only cFNB and intra-articular infiltration yielded better results than control group [standard mean difference=-0.68, 95% credible intervals (CrIs)=-1.03 to -0.33; standard mean difference=-0.53, 95% CrIs=-1.07 to -0.01, respectively]. Only cFNB exhibited better results with respect to morphine consumption day 2 after surgery (mean difference=-12.95, 95% CrIs=-19.70 to -6.53). CONCLUSIONS: Considering both pain score and morphine consumption, cFNB was potentially the most efficacious postoperative treatment for patients undergoing TKA.