Targeted serum proteome profiling reveals nicotinamide adenine dinucleotide phosphate (NADPH)-related biomarkers to discriminate linear IgA bullous disorder from dermatitis herpetiformis.

Linear IgA bullous dermatosis (LABD) and dermatitis herpetiformis (DH) represent the major subtypes of IgA mediated autoimmune bullous disorders. We sought to understand the disease etiology by using serum proteomics. We assessed 92 organ damage biomarkers in LAB, DH, and healthy controls using the Olink high-throughput proteomics. The positive proteomic serum biomarkers were used to correlate with clinical features and HLA type. Targeted proteomic analysis of IgA deposition bullous disorders vs. controls showed elevated biomarkers. Further clustering and enrichment analyses identified distinct clusters between LABD and DH, highlighting the involvement of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. Comparative analysis revealed biomarkers with distinction between LABD and DH and validated in the skin lesion. Finally, qualitative correlation analysis with DEPs suggested six biomarkers (NBN, NCF2, CAPG, FES, BID, and PXN) have better prognosis in DH patients. These findings provide potential biomarkers to differentiate the disease subtype of IgA deposition bullous disease.

L-arginine metabolism inhibits arthritis and inflammatory bone loss.

OBJECTIVES: To investigate the effect of the L-arginine metabolism on arthritis and inflammation-mediated bone loss. METHODS: L-arginine was applied to three arthritis models (collagen-induced arthritis, serum-induced arthritis and human TNF transgenic mice). Inflammation was assessed clinically and histologically, while bone changes were quantified by µCT and histomorphometry. In vitro, effects of L-arginine on osteoclast differentiation were analysed by RNA-seq and mass spectrometry (MS). Seahorse, Single Cell ENergetIc metabolism by profilIng Translation inHibition and transmission electron microscopy were used for detecting metabolic changes in osteoclasts. Moreover, arginine-associated metabolites were measured in the serum of rheumatoid arthritis (RA) and pre-RA patients. RESULTS: L-arginine inhibited arthritis and bone loss in all three models and directly blocked TNFα-induced murine and human osteoclastogenesis. RNA-seq and MS analyses indicated that L-arginine switched glycolysis to oxidative phosphorylation in inflammatory osteoclasts leading to increased ATP production, purine metabolism and elevated inosine and hypoxanthine levels. Adenosine deaminase inhibitors blocking inosine and hypoxanthine production abolished the inhibition of L-arginine on osteoclastogenesis in vitro and in vivo. Altered arginine levels were also found in RA and pre-RA patients. CONCLUSION: Our study demonstrated that L-arginine ameliorates arthritis and bone erosion through metabolic reprogramming and perturbation of purine metabolism in osteoclasts.

Ultrasound study of right ventricular myocardial perfusion and functional changes in hypertrophic cardiomyopathy.

BACKGROUND: To evaluate the changes of right ventricular (RV) myocardial perfusion and function in patients with hypertrophic cardiomyopathy (HCM) by myocardial contrast echocardiography (MCE) and speckle tracking (2D-STE), and to explore the relationship between RV myocardial perfusion and strain. METHODS: Conventional ultrasound, MCE and 2D-STE were performed on 29 HCM patients and 21 healthy subjects to analyze RV myocardial perfusion, RV global strain, RV free wall strain, and strain of each segment. The correlation between RV myocardial perfusion and strain was further analyzed in HCM patients. RESULTS: MCE results showed that the regional myocardial perfusion of the RV in HCM patients was decreased. Compared with the normal control group, the mean slope (ß) in the middle and apical segments of the RV free wall, and the peak intensity (A), ß, myocardial blood flow (MBF) of the ventricular septum decreased in HCM patients (P < 0.05). RV function was impaired in HCM patients. The RV global strain (RV GLS), and the strain of RV free wall and each segment were lower than those in the normal control group (P < 0.05). Correlation analysis showed that there was a certain correlation between RV myocardial perfusion and strain, such as the ß of the whole RV in HCM group had a positive correlation with the strain of the middle segment of the interventricular septum (r = 0.550, P = 0.002). CONCLUSIONS: The regional myocardial perfusion and strain of the RV in HCM patients are reduced, and there is a positive correlation between them, suggesting that the reduction of myocardial strain may be related to the impairment of myocardial microcirculation.

Targeting PI3K/AKT/mTOR pathway to enhance the anti-leukemia efficacy of venetoclax.

BACKGROUND: The treatment of acute myeloid leukemia (AML) is developing towards "targeted therapy", which faces challenges such as low sensitivity and drug resistance. Therefore, targeted drugs need to be used in combination with other drugs to overcome clinical problems. OBJECTIVE: AML cells and animal models were used to determine the synergistic anti-leukemic effect of Dactolisib (BEZ235) and Venetoclax (ABT199) and explore its mechanism. METHODS: In vitro experiments, we used cell counting kit-8 (CCK8), flow cytometry, real-time quantitative PCR (qPCR), and Western blot to detect the anti-leukemic effects of ABT199 and BEZ235. In vivo experiments, female nude mice were injected subcutaneously with THP-1 cells to form tumors, evaluate the combined effect of ABT199 and BEZ235 by indicators such as tumor size, tumor weight, Ki67 and cleaved-Caspase3 staining. The mice's body weight and HE staining were used to evaluate the liver injury and adverse drug reactions. RESULTS: The combination of BEZ235 and ABT199 has a synergistic effect through promoting apoptosis and inhibiting proliferation. The BEZ235 increased the drug sensitivity of ABT199 by reducing the MCL-1 protein synthesis and promoted the degradation of MCL-1 protein, which is considered as the mechanism of reversing ABT199 resistance. Furthermore, the BEZ235 and ABT199 can synergistically enhance the inhibition of PI3K/AKT/mTOR pathway. CONCLUSION: The combination of BEZ235 and ABT199 exhibits a synergistic anti-tumor effect in AML by down-regulating MCL-1 protein.

Application value of tumor necrosis factor inhibitors in in vitro fertilization-embryo transfer in infertile women with polycystic ovary syndrome.

BACKGROUND: Clinical value of tumor necrosis factor (TNF) inhibitors in in vitro fertilization-embryo transfer (IVF-ET) in infertile women with polycystic ovary syndrome (PCOS) was investigated in this study. METHODS: A retrospective analysis was performed on the clinical data of 100 PCOS patients who received IVF-ET for the first time at Hebei Institute of reproductive health science and technology from January 2010 to June 2020. The patients were divided into Inhibitor group and Control group according to whether they were treated with or without TNF inhibitors. Next, the two groups were subject to comparison in terms of the days of gonadotropin (Gn) use, total dosage of Gn, trigger time, hormone level and endometrial condition on the day of human chorionic gonadotropin (HCG) injection, the effects of two different regimens on controlled ovarian hyperstimulation (COH) and pregnancy outcomes. RESULTS: There were no significant differences in baseline characteristics between the two groups, including age, duration of infertility, body mass index (BMI), ovarian volume, antral follicle count, and basal hormone levels. Compared with the Control group, the days of Gn use and trigger time of patients in the Inhibitor group were significantly shortened, and the total Gn dosage was notably reduced. In terms of sex hormone levels on the HCG injection, the Inhibitor group displayed much lower serum estradiol levels while higher serum luteinizing hormone and progesterone (P) levels than the Control group. Notably, the high-quality embryo rate was also significantly increased with the use of TNF inhibitors. However, significant differences were not observed in endometrial thickness (on the day of HCG injection), proportion of endometrial A, B and C morphology (on the day of HCG injection), cycle cancellation rate, number of oocytes retrieved, fertilization rate, and cleavage rate between the two groups. Importantly, the clinical pregnancy rate in the Inhibitor group was significantly higher than that in the Control group, but there was no significant difference in the biochemical pregnancy rate, early abortion rate, multiple birth rate, ectopic pregnancy rate and number of live births between the two groups. CONCLUSION: Collectively, after application of TNF-α inhibitor regimen, superior overall treatment effect can be observed in infertile PCOS patients receiving IVF-ET. Therefore, TNF inhibitors have certain application value in IVF-ET in infertile women with PCOS.

Tabersonine attenuates obesity-induced renal injury via inhibiting NF-κB-mediated inflammation.

Obesity-induced metabolic disorders can cause chronic inflammation in the whole body, activating the nuclear factor kappa B (NF-κB) pathway and inducing apoptosis. Therefore, anti-inflammatory strategies may be effective in preventing obesity-related renal injury. Tabersonine (Tab) has been used pharmacologically to alleviate inflammation-related symptoms. This study evaluated the therapeutic effect of Tab on obesity-related renal injury and explored the pharmacological mechanism. Tab (20 mg/kg) relieved HFD-induced renal structural disorder and alleviated renal functional decline in mice, including improvement of renal tissue fibrosis, reducing renal cell apoptosis and inflammation in renal tissues. Mechanistically, we demonstrated that Tab inhibited the activation of NF-κB signaling pathway both in vivo and in vitro, thereby improving the renal tissue lesions in the mice with obesity-related renal injury. In both the obese mouse model and the mouse glomerular mesangial cell model, the natural compound Tab ameliorated HFD- and saturated fatty acid-induced renal cell injury by inhibiting the activation of NF-κB signaling pathway. Our data suggest that Tab may become a potential candidate for the prevention and treatment of obesity-related renal injury.

Effect of screw speed, temperature and moisture on physicochemical properties of corn gluten meal extrudate.

BACKGROUND: Corn gluten meal (CGM) is the main by-product of corn starch with rich protein and dietary fiber. The extrusion of CGM with a twin-screw extruder aimed to expand the novel utilization of this plant-protein resource. The impacts of screw speed, extrusion temperature, and material moisture on physicochemical properties of the extrudates were assessed. RESULTS: The microstructure depicted a favorable fiber-like structure formed under screw speed 120-150 rpm, extrusion temperature 140-150 °C, and material moisture 40-45%. Expansion ratio, rehydration ratio, water solubility index, hardness, and chewiness increased until screw speed reached 120 rpm. With accelerating extrusion temperature, these indicators showed an overall increasing trend. As for material moisture, expansion ratio, hardness, and chewiness showed a decreasing trend. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed that disulfide bonds were necessary for protein crosslinking during extrusion. CONCLUSION: It can be concluded that CGM is extrudable, whose textural and physicochemical properties vary as functions of the extruding parameters, providing diversity for its potential applications. © 2023 Society of Chemical Industry.

Factors Influencing Professional Identity of Psychiatric Nurses as Second Victims: A Cross-Sectional Study.

The current study aimed to explore the status and influencing factors of professional identity among psychiatric nurses as second victims in China by using a cross-sectional design. We investigated 291 psychiatric nurses from two psychiatric hospitals. Participants were asked to complete a demographic questionnaire, Second Victim Experience and Support Scale, Multidimensional Health Locus of Control Scale, and Professional Identity Scale for Nurses. Scores of professional identity of psychiatric nurses as second victims were moderate. Regression analysis showed that the second victim experience and support and internal control were significant predictors, explaining 34.2% of the variance in professional identity. Identifying risk factors related to the professional identity of psychiatric nurses as second victims will help managers take timely preventive measures to improve the awareness of the self-health responsibility of psychiatric nurses and reduce the adverse effects of patient safety incidents to enhance their professional identity. [Journal of Psychosocial Nursing and Mental Health Services, 61(12), 47-54.].

Effect of nimodipine combined with atorvastatin calcium on microinflammation and oxidative stress levels in patients with cerebral vasospasm after subarachnoid hemorrhage.

Objective: To evaluate the effect of nimodipine combined with atorvastatin calcium on the micro inflammation and oxidative stress levels in patients with cerebral vasospasm (CVS) after subarachnoid hemorrhage (SAH) and its clinical implications. Methods: A total of 80 patients with CVS caused by SAH who had been admitted to Baoding First Central Hospital from August 2021 to August 2022 were selected and randomly divided into two groups. The control group underwent conventional symptomatic treatment, while the experimental group was administered nimodipine combined with atorvastatin calcium on the basis of conventional treatment. The changes in the micro inflammatory cytokines and oxidative stress factors in the two groups were compared, as well as the differences in clinical efficacy and incidence of adverse drug reactions. Result: After treatment, the levels of inflammatory cytokines in the experimental group decreased more significantly than those in the control group (p=0.00). After treatment, the serum levels of oxidative stress factors were obviously higher in the experimental group than in the control group (p=0.00). After treatment, the total efficacy was 77.5% in the experimental group and 55% in the control group, and the difference was statistically significant (p=0.04). Conclusions: Nimodipine combined with atorvastatin calcium could significantly improve the clinical symptoms in patients with CVS after SAH, which would be beneficial, safe, and effective for the patient's recovery.

Epithelial HIF2α expression induces intestinal barrier dysfunction and exacerbation of arthritis.

OBJECTIVE: To investigate how the mucosal barrier in the intestine influences the development of arthritis, considering that metabolic changes in the intestinal epithelium influence its barrier function. METHODS: Intestinal hypoxia inducible factor (HIF)-2α expression was assessed before, at onset and during experimental arthritis and human rheumatoid arthritis (RA). Intestinal epithelial cell-specific HIF2α conditional knock-out mice were generated (HIF2α∆IEC) and subjected to collagen-induced arthritis. Clinical and histological courses of arthritis were recorded; T-cell and B-cell subsets were analysed in the gut and secondary lymphatic organs; and intestinal epithelial cells were subjected to molecular mRNA sequencing in HIF2α∆IEC and littermate control mice. The gut intestinal HIF2α target genes were delineated by chromatin immunoprecipitation and luciferase experiments. Furthermore, pharmacological HIF2α inhibitor PT2977 was used for inhibition of arthritis. RESULTS: Intestinal HIF2α expression peaked at onset of experimental arthritis and RA. Conditionally, deletion of HIF2α in gut epithelial cells inhibited arthritis and was associated with improved intestinal barrier function and less intestinal and lymphatic Th1 and Th17 activation. Mechanistically, HIF2α induced the transcription of the pore-forming claudin (CLDN)-15, which inhibits intestinal barrier integrity. Furthermore, treatment with HIF2α inhibitor decreased claudin-15 expression in epithelial cells and inhibited arthritis. CONCLUSION: These findings show that the HIF2α-CLDN15 axis is critical for the breakdown of intestinal barrier function at onset of arthritis, highlighting the functional link between intestinal homeostasis and arthritis.

The RNA-binding protein HuR regulates intestinal epithelial restitution by modulating Caveolin-1 gene expression.

The integrity of the intestinal mucosal barrier protects hosts against pathological conditions. Early mucosal restitution after wounding refers to epithelial cell migration into a defect. The RNA-binding protein HuR plays an important role in the posttranscriptional regulation of gene expression and is involved in many aspects of cellular physiology. In the present study, we investigated the role of HuR in the regulation of cell migration through the posttranscriptional regulation of Caveolin-1 (Cav-1). Online software was used to identify Cav-1 mRNA as a potential target of HuR. The interaction of HuR with Cav-1 mRNA was investigated via ribonucleoprotein immunoprecipitation (RNP IP) assays and biotin pulldown analysis. HuR was found to bind specifically to the Cav-1 3'-UTR rather than the coding region or 5'-UTR. Transfection of cells with siHuR decreased both HuR protein levels and Cav-1 protein levels; conversely, ectopic overexpression of HuR via infection of cells with an adenoviral vector containing HuR cDNA (AdHuR) increased Cav-1 protein levels without disturbing Cav-1 mRNA levels. Thus, HuR enhanced Cav-1 expression in vitro by stimulating Cav-1 translation. Intestinal epithelium-specific HuR knockout in mice decreased Cav-1 protein levels without changing Cav-1 mRNA levels, consistent with the in vitro results. Decreasing the levels of HuR via siHuR transfection inhibited early epithelial repair, but this effect was reversed by ectopic overexpression of GFP-tagged Cav-1. These results indicate that posttranscriptional regulation of Cav-1 gene expression by HuR plays a critical role in the regulation of rapid epithelial repair after wounding.

3D Printing Soft Matters and Applications: A Review.

The evolution of nature created delicate structures and organisms. With the advancement of technology, especially the rise of additive manufacturing, bionics has gradually become a popular research field. Recently, researchers have concentrated on soft robotics, which can mimic the complex movements of animals by allowing continuous and often responsive local deformations. These properties give soft robots advantages in terms of integration and control with human tissue. The rise of additive manufacturing technologies and soft matters makes the fabrication of soft robots with complex functions such as bending, twisting, intricate 3D motion, grasping, and stretching possible. In this paper, the advantages and disadvantages of the additive manufacturing process, including fused deposition modeling, direct ink writing, inkjet printing, stereolithography, and selective laser sintering, are discussed. The applications of 3D printed soft matter in bionics, soft robotics, flexible electronics, and biomedical engineering are reviewed.

Integrated Methylome and Transcriptome Analysis Provides Insights into the DNA Methylation Underlying the Mechanism of Cytoplasmic Male Sterility in Kenaf (Hibiscus cannabinus L.).

Cytoplasmic male sterility (CMS) is widely exploited in hybrid seed production. Kenaf is an important fiber crop with high heterosis. The molecular mechanism of kenaf CMS remains unclear, particularly in terms of DNA methylation. Here, using the anthers of a kenaf CMS line (P3A) and its maintainer line (P3B), comparative physiological, DNA methylation, and transcriptome analyses were performed. The results showed that P3A had considerably lower levels of IAA, ABA, photosynthetic products and ATP contents than P3B. DNA methylome analysis revealed 650 differentially methylated genes (DMGs) with 313 up- and 337 down methylated, and transcriptome analysis revealed 1788 differentially expressed genes (DEGs) with 558 up- and 1230 downregulated genes in P3A compared with P3B. Moreover, 45 genes were characterized as both DEGs and DMGs, including AUX,CYP, BGL3B, SUS6, AGL30 and MYB21. Many DEGs may be regulated by related DMGs based on methylome and transcriptome studies. These DEGs were involved in carbon metabolism, plant hormone signal transduction, the TCA cycle and the MAPK signaling pathway and were shown to be important for CMS in kenaf. These results provide new insights into the epigenetic mechanism of CMS in kenaf and other crops.

Effect of enamel-surface modifications on shear bond strength using different adhesive materials.

BACKGROUND: This study aimed to investigate the effect of enamel-surface modifications on the shear bond strength between ceramic brackets bonded using different adhesive materials and the enamel surface and to identify the most suitable clinical adhesive and bonding method. Whether the non-acid-etching treatment met the clinical bond strength was also determined. METHODS: A total of 108 extracted premolars were divided into nine groups (n = 12) based on the different enamel-surface modification techniques (acid etching, deproteinization, and wetting). Group 1 was bonded with Transbond™ XT adhesive, whereas groups 2-9 were bonded with resin-modified glass ionomer cement (RMGIC). The treatment methods for each group were as follows: groups 1 and 2, acid etching; group 3, acid etching and wetting; group 4, acid etching and deproteinization; group 5, acid etching, deproteinization, and wetting; group 6, deproteinization; group 7, deproteinization and wetting; group 8, without treatment; and group 9, wetting. The samples' shear bond strength was measured using an universal testing machine. Adhesive remnant index (ARI) was examined using a stereomicroscope. The enamel-surface morphology was observed with a scanning electron microscope. One-way ANOVA with Tukey's post-hoc test and chi-square test were used for statistical analysis, and p < 0.05 and α = 0.05 were considered statistically significant. RESULTS: The ARIs of groups 1-5 and 6-9 were statistically significant (p = 0.000). The enamel surface of groups 1-5 was demineralized, and only a tiny amount of protein remained in groups 7 and 8, whereas a thick layer of protein remained in groups 8 and 9. CONCLUSIONS: RMGIC adhesive did not damage the enamel surface and achieved the required clinical bond strength. The enamel surface was better treated with 5.25% sodium hypochlorite preferably under non-acid-etching conditions.

Effects of pore size and porosity on cytocompatibility and osteogenic differentiation of porous titanium.

To find out the optimal porosity and pore size of porous titanium (Ti) regarding the cytocompatibility and osteogenic differentiation. Six groups of porous Ti samples with different porosities and pore sizes were fabricated by the powder metallurgy process. The microstructure and compressive mechanical properties were characterized. The cytocompatibility was examined by a series of biological tests as protein absorption with BCA assay kit, cell attachment with laser scanning confocal microscopy and vinculin expression, cell proliferation with CCK-8 assay. Cell differentiation and calcification were detected by qPCR and Alizarin Red S dying respectively. Pores distributed homogeneously throughout the porous Ti samples. The compressive test results showed that Young's modulus ranged from 2.80 ± 0.03 GPa to 5.43 ± 0.34 GPa and the compressive strength increased from 112.4 ± 3.6 MPa to 231.1 ± 9.4 MPa. Porous Ti with high porosity (53.3 ± 1.2%) and small pore size (191.6 ± 3.7 µm) adsorbed more proteins. More MC3T3-E1 cells adhered onto dense Ti samples than onto any other porous ones already after culture and no difference was identified within the porous groups. The porous structure of porous Ti with a porosity of 53.3 ± 1.2% and an average pore size of 191.6 ± 3.7 µm facilitated cell differentiation and calcification. Small pores were not beneficial to the osteo-initiation at the very beginning. Porous Ti with a porosity of 53.3 ± 1.2% and an average pore size of 191.6 ± 3.7 µm fabricated by powder metallurgy process showed the expected mechanical property and improved osseointegration as implants in dental treatment.

High Accurate Environmental Sound Classification: Sub-Spectrogram Segmentation versus Temporal-Frequency Attention Mechanism.

In the important and challenging field of environmental sound classification (ESC), a crucial and even decisive factor is the feature representation ability, which can directly affect the accuracy of classification. Therefore, the classification performance often depends to a large extent on whether the effective representative features can be extracted from the environmental sound. In this paper, we firstly propose a sub-spectrogram segmentation with score level fusion based ESC classification framework, and we adopt the proposed convolutional recurrent neural network (CRNN) for improving the classification accuracy. By evaluating numerous truncation schemes, we numerically figure out the optimal number of sub-spectrograms and the corresponding band ranges, and, on this basis, we propose a joint attention mechanism with temporal and frequency attention mechanisms and use the global attention mechanism when generating the attention map. Finally, the numerical results show that the two frameworks we proposed can achieve 82.1% and 86.4% classification accuracy on the public environmental sound dataset ESC-50, respectively, which is equivalent to more than 13.5% improvement over the traditional baseline scheme.

The Chemical Compositions, and Antibacterial and Antioxidant Activities of Four Types of Citrus Essential Oils.

Nanfeng mandarins (Citrus reticulata Blanco cv. Kinokuni), Xunwu mandarins (Citrus reticulata Blanco), Yangshuo kumquats (Citrus japonica Thunb) and physiologically dropped navel oranges (Citrus sinensis Osbeck cv. Newhall) were used as materials to extract peel essential oils (EOs) via hydrodistillation. The chemical composition, and antibacterial and antioxidant activities of the EOs were investigated. GC-MS analysis showed that monoterpene hydrocarbons were the major components and limonene was the predominate compound for all citrus EOs. The antibacterial testing of EOs against five different bacteria (Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Salmonella typhimurium) was carried out using the filter paper method and the broth microdilution method. Kumquat EO had the best inhibitory effect on B. subtilis, E. coli and S. typhimurium with MIC (minimum inhibitory concentration) values of 1.56, 1.56 and 6.25 µL/mL, respectively. All citrus EOs showed the antioxidant activity of scavenging DPPH and ABTS free radicals in a dose-dependent manner. Nanfeng mandarin EO presented the best antioxidant activity, with IC50 values of 15.20 mg/mL for the DPPH assay and 0.80 mg/mL for the ABTS assay. The results also showed that the antibacterial activities of EOs might not be related to their antioxidant activities.

Predictive value of DWI-FLAIR Mismatch in patients with Ischemic Stroke and receiving Endovascular treatment beyond Time Window.

OBJECTIVE: To investigate the efficacy and safety of endovascular treatment in patients having acute ischemic stroke with over-time window under DWI-FLAIR mismatch. METHODS: From January 2018 to January 2020, 80 patients who met the research criteria in the First Central Hospital of Baoding, China were selected. According to the time of onset, they were divided into test group and control group, with 40 cases in each group. Forty patients in the test group were beyond time window (6~24h) and the MRI showed a DWI-FLAIR mismatch. Forty patients in the control group were within the time window (< 6h). All patients received endovascular treatment (EVT). The mRS, NIHSS and infarct volume of patients in the test group were compared and analyzed before and 30 and 90 days after treatment, as well as the indicators of both groups of patients before and after treatment, to determine therapeutic effect in patients receiving EVT beyond time window. Meanwhile, the recanalization of the blood vessel and the incidence of cerebral hemorrhage of patients in both groups were compared to determine the safety in patients receiving EVT beyond time window under DWI-FLAIR mismatch. RESULTS: The mRS, NIHSS and infarct size in the test group were significantly improved before and 30 and 90 days after treatment (p<0.05). The test group showed no significant difference in mRS, NIHSS and other indicators when compared with the control group (p>0.05). There was no significant difference in the rate of recanalization of the blood vessel and intracranial hemorrhage after treatment between both groups (p>0.05). CONCLUSION: DWI-FLAIR mismatch can be used as an objective imaging basis for intravascular interventional therapy in patients with stroke with over-time window and large vessel occlusion. It has the advantages of short examination time, non-invasiveness, no need for contrast agents, simple implementation, clear guidance.

Mutation topography and risk stratification for de novo acute myeloid leukaemia with normal cytogenetics and no nucleophosmin 1 (NPM1) mutation or Fms-like tyrosine kinase 3 internal tandem duplication (FLT3-ITD).

About 25% of patients with newly diagnosed acute myeloid leukaemia (AML) have normal cytogenetics and no nucleophosmin 1 (NPM1) mutation or Fms-like tyrosine kinase 3 internal tandem duplication (FLT3-ITD). The prognosis and best therapy for these patients is controversial. We evaluated 158 newly diagnosed adults with this genotype who achieved histological complete remission within two cycles of induction therapy and were assigned to two post-remission strategies with and without an allotransplant. Targeted regional sequencing at diagnosis was performed and data were used to estimate their prognosis, including relapse and survival. In multivariable analyses, having wild-type or mono-allelic mutated CCAAT/enhancer-binding protein alpha (CEBPA) [hazard ratio (HR) 2·39, 95% confidence interval (CI) 1·08-5·30; P = 0·032), mutated NRAS (HR 2·67, 95% CI 1·36-5·25; P = 0·004), mutated colony-stimulating factor 3 receptor (CSF3R) (HR 2·85, 95% CI 1·12-7·27; P = 0·028) and a positive measurable residual disease (MRD)-test after the second consolidation cycle (HR 2·88, 95% CI 1·32-6·30; P = 0·008) were independently correlated with higher cumulative incidence of relapse (CIR). These variables were also significantly associated with worse survival (HR 3·02, 95% CI 1·17-7·78, P = 0·022; HR 3·62, 95% CI 1·51-8·68, P = 0·004; HR 3·14, 95% CI 1·06-9·31, P = 0·039; HR 4·03, 95% CI 1·64-9·89, P = 0·002; respectively). Patients with ≥1 of these adverse-risk variables benefitted from a transplant, whereas the others did not. In conclusion, we identified variables associated with CIR and survival in patients with AML and normal cytogenetics without a NPM1 mutation or FLT3-ITD.

RNA-Binding Protein HuR Regulates Paneth Cell Function by Altering Membrane Localization of TLR2 via Post-transcriptional Control of CNPY3.

BACKGROUND & AIMS: Paneth cells secrete antimicrobial proteins including lysozyme via secretory autophagy as part of the mucosal protective response. The ELAV like RNA-binding protein 1 (ELAVL1, also called HuR) regulates stability and translation of messenger RNAs (mRNAs) and many aspects of mucosal physiology. We studied the posttranscriptional mechanisms by which HuR regulates Paneth cell function. METHODS: Intestinal mucosal tissues were collected from mice with intestinal epithelium (IE)-specific disruption of HuR (IE-HuR-/-), HuRfl/fl-Cre- mice (controls), and patients with inflammatory bowel diseases and analyzed by histology and immunohistochemistry. Paneth cell functions were determined by lysozyme-immunostaining assays. We isolated primary enterocytes from IE-HuR-/- and control mice and derived intestinal organoids. HuR and the chaperone CNPY3 were overexpressed from transgenes in intestinal epithelial cells (IECs) or knocked down with small interfering RNAs. We performed RNA pulldown assays to investigate interactions between HuR and its target mRNAs. RESULTS: Intestinal tissues from IE-HuR-/- mice had reduced numbers of Paneth cells, and Paneth cells had fewer lysozyme granules per cell, compared with tissues from control mice, but there were no effects on Goblet cells or enterocytes. Intestinal mucosa from patients with inflammatory bowel diseases had reduced levels of HuR and fewer Paneth cells. IE-HuR-/- mice did not have the apical distribution of TLR2 in the intestinal mucosa as observed in control mice. IECs from IE-HuR-/- mice expressed lower levels of CNPY3. Intestinal organoids from IE-HuR-/- mice were smaller and contained fewer buds compared with those generated from controls, and had fewer lysozyme-positive cells. In IECs, knockdown of HuR decreased levels of the autophagy proteins LC3-I and LC3-II, compared with control cells, and prevented rapamycin-induced autophagy. We found HuR to interact directly with the Cnpy3 mRNA coding region and increase levels of CNPY3 by increasing the stability and translation of Cnpy3 mRNA. CNPY3 bound TLR2, and cells with knockdown of CNPY3 or HuR lost membrane localization of TLR2, but increased cytoplasmic levels of TLR2. CONCLUSIONS: In studies of mice, IECs, and human tissues, we found HuR to increase expression of CNPY3 at the posttranscriptional level. CNPY3 is required for membrane localization of TLR2 and Paneth cell function.