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Chitosan (C) and zein (Z) were used to develop bilayer films with a characteristic one-way water barrier using a layer-by-layer (LBL) casting method. The effects of mass ratios (C:Z

The promoting role that miR-18a-3p plays in osteoporosis (OP) has been previously described. However, the detailed mechanisms remain unclear. Bone tissues were collected from healthy patients, OP patients, and patients with osteoporotic spinal fractures. An osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) was constructed to detect the expression of miR-18a-3p and glutamate AMPA receptor subunit 1 (GRIA1). Alkaline phosphatase (ALP) activity and a qRT-PCR analysis were used to detect ALP content, alizarin red S staining was used to detect calcium deposition, and qRT-PCR was used to evaluate runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), and osteopontin (OPN) expression levels. A dual-luciferase reporter and RNA pull-down assay was used to verify the targeted correlation between miR-18a-3p and GRIA1. We observed an increase in miR-18a-3p expression and a decrease in GRIA1 expression in OP and osteoporotic vertebral fracture patients. Upregulation of miR-18a-3p restrained the activity and expression of ALP in hBMSCs, inhibited the expression of RUNX2, OCN, and OPN, and inhibited calcium deposition. Knockdown of miR-18a-3p or upregulation of GRIA1 promoted osteogenic differentiation. Our findings indicate that miR-18a-3p promotes OP progression by regulating GRIA1 expression, suggesting that targeting miR-18a-3p/GRIA1 may be a therapeutic strategy for OP.

This paper aims to clarify the mechanisms and driving factors of subgrade mud pumping for ballastless track in high-speed railway. A full-scale model test system in which the train dynamic loading ...

The long-term loading of the trains on the subgrade in service is the cyclic loading when the train passes and the intermittent stage when there is no train passing. However, in most studies, continuous loading was used to investigate the dynamic characteristics of a subgrade soil, which ignores the effect of loading intermittence. The aims of this research were first to study the cumulative permanent deformation behaviors under intermittent cyclic loading of trains, then to establish the classification criterion for cumulative permanent strain behaviors, and finally to propose a practical method for estimating the critical dynamic stress in the intermittent loading mode. For this purpose, according to the actual conditions of the subgrade of Shuo-Huang Heavy-Haul Railway, the monotonic triaxial test and the cyclic triaxial test with intermittent loading were performed. Based on the shakedown theory, the cumulative permanent strain behaviors under intermittent loading can be classified into three categories of plastic shakedown, plastic creep, and incremental collapse. Then, the classification criterion was established based on the permanent strain rate. Finally, an empirical formula of critical dynamic stress for intermittent loading, which could also consider the varying moisture content, was proposed.

Background Atelectasis is a common postoperative complication. Peri-operative lung protection can reduce atelectasis; however, it is not clear whether this persists into the postoperative period. Objective To evaluate to what extent lung-protective ventilation reduces peri-operative atelectasis in children undergoing nonabdominal surgery. Design Randomised, controlled, double-blind study. Setting Single tertiary hospital, 25 July 2019 to 18 January 2020. Patients A total of 60 patients aged 1 to 6 years, American Society of Anesthesiologists physical status 1 or 2, planned for nonabdominal surgery under general anaesthesia (≤2 h) with mechanical ventilation. Interventions The patients were assigned randomly into either the lung-protective or zero end-expiratory pressure with no recruitment manoeuvres (control) group. Lung protection entailed 5 cmH2O positive end-expiratory pressure and recruitment manoeuvres every 30 min. Both groups received volume-controlled ventilation with a tidal volume of 6 ml kg-1 body weight. Lung ultrasound was conducted before anaesthesia induction, immediately after induction, surgery and tracheal extubation, and 15 min, 3 h, 12 h and 24 h after extubation. Main outcome measures The difference in lung ultrasound score between groups at each interval. A higher score indicates worse lung aeration. Results Patients in the lung-protective group exhibited lower median [IQR] ultrasound scores compared with the control group immediately after surgery, 4 [4 to 5] vs. 8 [4 to 6], (95% confidence interval for the difference between group values -4 to -4, Z = -6.324) and after extubation 3 [3 to 4] vs. 4 [4 to 4], 95% CI -1 to 0, Z = -3.161. This did not persist from 15 min after extubation onwards. Lung aeration returned to normal in both groups 3 h after extubation. Conclusions The reduced atelectasis provided by lung-protective ventilation does not persist from 15 min after extubation onwards. Further studies are needed to determine if it yields better results in other types of surgery. Trial registration Chictr.org.cn (ChiCTR2000033469).

Objective The cardiotoxicity of doxorubicin (DOX) reduces the quality of life and prognosis of cancer patients, and therefore its clinical application has been largely restricted. This study aimed to assess the effects of cryptotanshione (CPT) on DOX-induced rat cardiac insufficiency. Results CPT treatment significantly suppressed apoptosis in vitro. The oral administration of CPT significantly improved cardiac function in the rat model, reduced collagen production and suppressed apoptosis and the production of reactive oxygen species in the heart tissue. Transcriptomic profiling and its relevant bioinformatics analysis showed that CPT suppressed doxorubicin-induced cardiotoxicity by inhibiting p53 signaling pathway. Conclusion Transcriptomic profiling and bioinformatics analysis can be used to evaluate the cardio-protective effect of CPT through inactivating p53 signaling pathway in the doxorubicin-mediated myocardial damage model. Methods F-actin staining and flow cytometry were used to assess the effects of CPT on cardiomyocytes. In vivo, echocardiography and hemodynamic evaluation were used to assess the effects of CPT on the cardiac dysfunction in rats. Furthermore, transcriptomic profiling and bioinformatics analysis, as well as western blot analysis, were used to determine that CPT induced changes in the signaling pathways in the model.

Since there is an interval time between two trains traveling in the same direction on an automatically blocked line, the dynamic loading on the railway subgrade is composed of periodic vibration when the train passes and an intermittent stage after a train has passed. Take into account this characteristic, a series of continuous-stopping repeated load triaxial tests were conducted to study deformation characteristics such as plastic strain and elastic strain of fine-grained soil under intermittent loading. Compared to continuous loading, it is found that the intermittent stage has a significant effect on the deformation characteristics of subgrade. The excess pore water pressure accumulated during the loading stage dissipated during the intermittent stage. The development of plastic strain in subsequent loading stages slowed down significantly, and the axial strain of the sample rebounded in the intermittent stage, so cumulative plastic strain was reduced. The intermittent effect on reducing elastic strain was, however, limited. An increase in intermittent time can also improve the resistance of the sample to subsequent cyclic loading. An alternating continuous-stopping loading mode can better simulate intermittent loading of trains in actual train operation and produce more practical results, providing a theoretical basis and guidance for further understanding of deformation characteristics of railway subgrade.

Fuzi (Aconitum carmichaelii Debx) has been traditionally used for the treatment of ulcerative colitis (UC) in China for thousands of years. The total alkaloids of A. carmichaelii (AAC) have been considered as the main medicinal components of fuzi, whereas its underlying anti-UC mechanisms remain elusive. In the present study, the dextran sulfate sodium (DSS)-induced UC mice model, which was consistent with the symptoms and pathological features of human UC, was established to comprehensively evaluate the anti-UC effects of AAC. The results indicated that AAC effectively improved the weight loss, disease activity index (DAI), spleen hyperplasia, and colon shortening, and thus alleviated the symptoms of UC mice. Meanwhile, AAC not only inhibited the MPO enzyme and the abnormal secretion of inflammatory cytokines (TNF-α, IL-1β, IL-6, IFN-γ, and IL-17A) and suppressed the overexpression of inflammatory mediators (TNF-α, IL-1β, and IL-6) of mRNA but also reduced the phosphorylation of p38 MAPK, ERK, and JNK, and the protein expressions of NF-κB, IκB-α, STAT3, and JAK2 in the colon tissue. Furthermore, the LC-MS/MS quantitative determination suggested that the three low toxic monoester alkaloids were higher in both contents and proportion than that of the three high toxic diester alkaloids. Additionally, molecular docking was hired to investigate the interactions between alkaloid-receptor complexes, and it suggested the three monoester alkaloids exhibited higher binding affinities with the key target proteins of MAPK, NF-κB, and STAT3. Our finding showcased the noteworthy anti-UC effects of AAC based on the MAPK/NF-κB/STAT3 signaling pathway, which would provide practical and edge-cutting background information for the development and utilization of A. carmichaelii as a potential natural anti-UC remedy.

Summary Autophagy is a conserved biological process that maintains cell homeostasis by targeting macromolecules for lysosome-mediated degradation. The levels of autophagy are relatively lower under normal conditions than under stress conditions (e.g., starvation), as autophagy is usually stimulated after multiple stresses. However, many autophagy-related regulators are still expressed under normal conditions. Although these regulators have been studied deeply in autophagy regulation, the nonautophagic roles of these regulators under normal conditions remain incompletely understood. Here, we found that autophagy-related 5 (ATG5), which is a key regulator of autophagy, regulates c-Myc protein degradation under normal conditions through the ubiquitin-proteasome pathway. We also found that ATG5 binds c-Myc and recruits the E3 ubiquitin-protein ligase FBW7 to promote c-Myc degradation. Moreover, ATG5-mediated degradation of c-Myc limits cell growth under normal conditions and is essential for embryonic stem cell differentiation. Therefore, this study reveals a nonautophagic role of ATG5 in regulating of c-Myc protein degradation., Graphical abstract, Highlights • ATG5 differentially regulates cell growth between normal and starvation conditions • ATG5 recruits FBW7 to regulate c-Myc protein degradation under normal conditions • ATG5-mediated degradation of c-Myc limits cell growth under normal conditions • ATG5 negatively regulates the protein level of c-Myc during ESC differentiation, Cell biology; Functional aspects of cell biology

This study aimed to reveal the effects of vacuum-impregnated carboxymethyl chitosan (CMCS) coating with pomegranate peel extract (PPE) on quality retention of fish flesh during refrigeration. Herein, CMCS-PPE coating was effective in attenuating quality loss of grass carp fillets. Compared to Control, the levels of drip loss, total volatile base nitrogen, and K value in coated samples were sharply decreased (p 0.05) by 24.5%, 35.3% and 25.2% on day 9, respectively. Meanwhile, the coating also helped inhibit oxidation, bioamine accumulation, and texture softening in fillets. Moreover, the microbial enumeration was reduced by1.4 lg cfu/g as compared to Control on day 6 afterward, and high throughput sequencing analysis further showed the active coating contributed to the notable growth suppression of spoilage bacteria like Shewanella. Additionally, the positive effect of the coating scheme was also verified in longsnout catfish and snakehead, further confirming its good applicability for fish flesh preservation.

Fuzi (

The dynamic response of the subgrade under moving train loads provides information on subgrade settlement prediction, condition evaluation, and so forth. This paper presents the field dynamics tests on the transition subgrade in the Shuo-Huang heavy-haul railway in China. The variation characteristics of the peak dynamic displacements along the track and subgrade slope were analyzed, and the random distribution characteristics of the peak dynamic displacements at the subgrade shoulder were studied. The response characteristics of the subgrade during the train passage were investigated, and the attenuation regularities of vibration along the subgrade slope were identified. The results indicated that the action of the train moving loads on the subgrade has obvious periodicity, and two bogies in the adjacent wagons should be considered as one loading unit. The peak dynamic displacements at the subgrade shoulder obey normal distribution under the repeated loading of the loading unit. The subgrade bed is dramatically influenced by the dynamic loadings of the trains, and the moving train loads have little influence on the part below the subgrade bed. The results of the research provide the basis for the evaluation of instantaneous and long-term dynamic stability of the subgrade and offer guidance for simulating train moving loads in the model test and numerical analysis to study the dynamic response of the subgrade.

Hederagenin (HED) is a bioactive natural compound of pentacyclic triterpenes extracted from many medicinal plants. It has a wide range of antitumor cytotoxic effects and significant anti-inflammation effects. However, at present, it is unclear whether HED can inhibit cardiac remodelling caused by diabetic cardiomyopathy. In this study, we evaluated the effects of HED on pathological abnormalities in cardiac structures and cardiac insufficiency caused by diabetic cardiomyopathy and focused on the inflammatory signalling pathways of the diabetic heart. Treatment with HED reduced pro-inflammatory cytokines, the heart and body mass of diabetic db/db mice but had no effect on fasting plasma glucose (FPG). Moreover, after HED treatment, the cardiac dysfunction of diabetic mice was relieved, and myocardial hypertrophy and fibrosis decreased. Furthermore, HED inhibited the nuclear translocation of nuclear factor-κB (NF-κB) and Smads and decreased the transcriptional activity of NF-κB and Smads. Additionally, the expression levels of transforming growth factor (TGF)-β1 and collagen I, which are target downstream molecules of the NF-κB and Smads signalling pathways, were also decreased in diabetic hearts. Taken together, our findings suggest that the cardioprotective effect of HED may be achieved by reducing the activation of inflammation-associated NF-κB and Smads signalling. We suggest that the protective effect of HED on the diabetic heart, as revealed in this study, should be further explored in-depth to elucidate its cell biology and molecular mechanisms.

The role of different spatial frequency bands in threat detection has been explored extensively. However, most studies use manual responses and the results are mixed. Here, we aimed to investigate the contribution of spatial frequency information to threat detection by using three response types, including manual responses, eye movements, and reaching movements, together with a priming paradigm. The results showed that both saccade and reaching responses were significantly faster to threatening stimuli than to nonthreatening stimuli when primed by low-spatial-frequency gratings rather than by high-spatial-frequency gratings. However, the manual response times to threatening stimuli were comparable to nonthreatening stimuli, irrespective of the spatial frequency content of the primes. The findings provide clear evidence that low-spatial-frequency information can facilitate threat detection in a response-specific manner, possibly through the subcortical magnocellular pathway dedicated to processing threat-related signals, which is automatically prioritized in the oculomotor system and biases behavior.

To investigate the influence of miR-18a-3p and ADCY5 on OP and osteogenic differentiation of human Mesenchymal stem cell (hBMSCs) and its possible mechanism. Samples were collected from osteoporotic patients with or without vertebral compression fracture, and without OP volunteers. MiR-18a-3p and ADCY5 mRNA expression levels in the tissue samples and hBMSCs during osteogenic differentiation were detected。MiR-18a-3p mimic and OE-ADCY5 were introduced into hBMSCs to research the effects of miR-18a-3p and ADCY5 on osteogenesis differentiation of hBMSCs. Dual luciferase reporter system and RNA pull-down were applied to determine whether ADCY5 was a target gene of miR-18a-3p. Compared with the control group, ADCY5 expression level was down-regulated in patients with OP-no-Frx and OP-Frx, but that of miR-18a-3p was up-regulated. In addition, ADCY5 increased during osteogenesis differentiation of hBMSCs, whereas miR-18a-3p did not. OE-ADCY5 significantly facilitated calcium deposition, ALP activity, osteoblast protein expression (OSX, ALP and EUNX2), miR-18a-3p mimic inhibited osteogenic differentiation, and partially reversed the effect of OE-ADCY5 on osteogenic differentiation. In general, miR-18a-3p targets ADCY5 to promote OP and may be involved in spinal fracturs.

Two phenazine compounds, diastaphenazine and izumiphenazine C, with complex structures and promising antitumour activity have been isolated from the plant endophytic actinomycete Streptomyces diastaticus W2. Their putative biosynthetic gene cluster (dap) was identified by heterologous expression and gene knockout. There are twenty genes in the dap cluster. dap14-19 related to shikimic pathway were potentially involved in the precursor chorismic acid biosynthesis, and dapBCDEFG were confirmed to be responsible for the biosynthesis of the dibenzopyrazine ring, the nuclear structure of phenazines. Two transcriptional regulatory genes dapR and dap4 played the positive regulatory roles on the phenazine biosynthetic pathway. Most notably, the dimerization of the dibenzopyrazine ring in diastaphenazine and the loading of the complex side chain in izumiphenazine C could be catalysed by the cyclase homologous gene dap5, suggesting an unusual modification strategy tailoring complex phenazine biosynthesis. Moreover, metabolite analysis of the gene deletion mutant strain S. albus::23C5Δdap2 and substrate assay of the methyltransferase Dap2 clearly revealed the biosynthetic route of the complex side chain in izumiphenazine C.

Autophagy is a conserved biological process that maintains cell homeostasis mainly by targeting macromolecules or organelles for lysosome-mediated degradation. The levels of autophagy are relatively lower under normal conditions than under stress conditions (e.g., starvation or infection), as autophagy is usually stimulated under such stress conditions. However, many autophagy-related regulators are still expressed under normal conditions. Although these regulators have been studied in depth in the context of autophagy regulation, the nonautophagic roles of these regulators under normal conditions remain incompletely understood. Here, we found that autophagy-related 5 (ATG5), which is a key regulator of autophagy, regulates c-Myc protein degradation under normal conditions through the ubiquitin-proteasome pathway. We also found that ATG5 binds c-Myc and recruits the E3 ubiquitin-protein ligase FBW7 to promote c-Myc protein degradation. Moreover, ATG5-mediated regulation of c-Myc limits cell growth under normal conditions and is essential for embryonic stem cell differentiation. Therefore, this study reveals a novel nonautophagic role of ATG5 in the regulation of c-Myc protein degradation.

Deformation of the subgrade bed layer under cyclic dynamic loading is one of the key factors to evaluate the operating performance of subgrade. To study the stress–strain characteristics of coarse-grained soil (CGS) under cyclic loading, a series of large-scale dynamic triaxial tests were carried out at different confining pressures and dynamic stress amplitudes. Results show that the variation of soil stiffness is closely related to cyclic number and confining pressure under cyclic loading. Backbone curve model is developed regarding confining pressure and cyclic number. Compared with the traditional backbone curve model, the proposed model can reflect the CGS stiffness hardening with the increasing of cyclic number and be used to determine the dynamic strength of CGS when the dynamic strain is less than the dynamic yield strain and to provide a reference for evaluating dynamic stress deformation stability for subgrade bed and design.

Summary Histones are the main components of chromatin, and the protein levels of histones significantly affect chromatin assembly. Here, we describe detailed protocols for histone H2B purification from bacteria and for the separation of nucleolar fractions and cytoplasmic and nucleoplasmic fractions. Finally, the in vitro ubiquitination and degradation of H2B by distinct cellular fractions are described. For complete details on the use and execution of this protocol, please refer to Liu et al. (2021)., Graphical abstract, Highlights • Protocols to purify nucleoli, cytoplasm, and nucleoplasm • In vitro H2B degradation and ubiquitination assays by these cellular components • In vitro ubiquitination and degradation of bacterial histone H2B by the nucleoli fractions, Histones are the main components of chromatin, and the protein levels of histones significantly affect chromatin assembly. Here, we describe detailed protocols for histone H2B purification from bacteria and for the separation of nucleolar fractions and cytoplasmic and nucleoplasmic fractions. Finally, the in vitro ubiquitination and degradation of H2B by distinct cellular fractions are described.

Fuzi and ganjiang are widely used as traditional Chinese medicines (TCM) in China, Korea, Japan, and many other southeast Asian countries for treating ulcerative colitis (UC), emesis and heart failure for more than 1800 years. However, the underlying mechanism of fuzi, ganjiang and fuzi-ganjiang herb pair is still unclear. In our study, we explored the therapeutic effects of fuzi, ganjiang and fuzi-ganjiang herb pair against dextran sulfate sodium (DSS)-induced UC in mice model, along with the relevant mechanism.The contents of each marker compound in fuzi decoction (FD), ganjiang decoction (GD) and fuzi-ganjiang decoction (FGD) were determined using LC-MS/MS. During the experiment, bodyweight changes in each group were monitored every 5 days. On the day of sacrifice, colonic length, disease activity index (DAI) and spleen weight were also evaluated and histopathological examination was performed through hematoxylineosin (HE) staining. The levels of myeloperoxidase (MPO) and inflammatory cytokines in colon tissues were determined by enzyme-linked immunosorbent assay (ELISA), and then the relative mRNA productions of inflammatory mediators, such as MPO, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 were measured by real-time polymerase chain reaction (PCR). Involvement of MAPK, STAT3 and NF-κB signaling pathways in the pathogenesis of UC was determined in each group using Western Blot (WB) analysis.Compared with fuzi and ganjiang single decoction, the content of the alkaloids derived from fuzi (especially the diester alkaloid with strong toxicity, hypaconitine) in fuzi-ganjiang herb pair decoction was reduced. Additionally, the 6-gingerol, which was not found in ganjiang single decoction, was retained in fuzi-ganjiang herb pair decoction. FD, GD, and FGD significantly restored the bodyweight reduction, colon shortening, DAI elevation, splenomegaly and histological score in DSS-induced UC mice. Furthermore, except for the failure of low dosage of ganjiang decoction (GD-L) on IL-17A, all FD, GD and FGD significantly inhibited the production of MPO and inflammatory cytokines, such as IFN-γ, TNF-α, IL-1β, IL-6, IL-10 and IL-17A, and suppressed the relative expression of inflammatory mediators, such as MPO, iNOS and COX-2 mRNA in colon tissues of DSS-induced mice. According to WB analysis, fuzi, ganjiang and fuzi-ganjiang combination inhibited the activation of MAPK, NF-κB and STAT3 signaling pathways.Our study demonstrated that fuzi, ganjiang and fuzi-ganjiang combination possess prominent anti-inflammatory activities against DSS-induced UC mice; the involved mechanism may be related to inhibition the activation of MAPK, NF-κB, and STAT3 signaling pathways.

Objective: The cardiotoxicity of doxorubicin (DOX) reduces the quality of life and prognosis of cancer patients, and therefore its clinical application has been largely restricted. This study aimed to assess the effects of cryptotanshione (CPT) on DOX-induced rat cardiac insufficiency. Results: CPT treatment significantly suppressed apoptosis in vitro. The oral administration of CPT significantly improved cardiac function in the rat model, reduced collagen production and suppressed apoptosis and the production of reactive oxygen species in the heart tissue. Transcriptomic profiling and its relevant bioinformatics analysis showed that CPT suppressed doxorubicin-induced cardiotoxicity by inhibiting p53 signaling pathway. Conclusion: Transcriptomic profiling and bioinformatics analysis can be used to evaluate the cardio-protective effect of CPT through inactivating p53 signaling pathway in the doxorubicin-mediated myocardial damage model. Methods: F-actin staining and flow cytometry were used to assess the effects of CPT on cardiomyocytes. In vivo, echocardiography and hemodynamic evaluation were used to assess the effects of CPT on the cardiac dysfunction in rats. Furthermore, transcriptomic profiling and bioinformatics analysis, as well as western blot analysis, were used to determine that CPT induced changes in the signaling pathways in the model.

Hederagenin (HED) is a bioactive natural compound of pentacyclic triterpenes extracted from many medicinal plants. It has a wide range of antitumor cytotoxic effects and significant anti-inflammation effects. However, at present, it is unclear whether HED can inhibit cardiac remodelling caused by diabetic cardiomyopathy. In this study, we evaluated the effects of HED on pathological abnormalities in cardiac structures and cardiac insufficiency caused by diabetic cardiomyopathy and focused on the inflammatory signalling pathways of the diabetic heart. Treatment with HED reduced pro-inflammatory cytokines, the heart and body mass of diabetic db/db mice but had no effect on fasting plasma glucose (FPG). Moreover, after HED treatment, the cardiac dysfunction of diabetic mice was relieved, and myocardial hypertrophy and fibrosis decreased. Furthermore, HED inhibited the nuclear translocation of nuclear factor-κB (NF-κB) and Smads and decreased the transcriptional activity of NF-κB and Smads. Additionally, the expression levels of transforming growth factor (TGF)-β1 and collagen I, which are target downstream molecules of the NF-κB and Smads signalling pathways, were also decreased in diabetic hearts. Taken together, our findings suggest that the cardioprotective effect of HED may be achieved by reducing the activation of inflammation-associated NF-κB and Smads signalling. We suggest that the protective effect of HED on the diabetic heart, as revealed in this study, should be further explored in-depth to elucidate its cell biology and molecular mechanisms.

Ethnopharmacological relevance Qingxue jiedu Formulation (QF) is composed of two classic prescriptions which have been clinically used for more than 5 centuries and appropriately modified through basic theory of traditional Chinese medicine for treating various skin inflammation such as atopic dermatitis (AD), acute dermatitis and rash. Although QF possesses a prominent clinical therapeutic effect, seldom pharmacological studies on its anti-AD activity are conducted. Aim of the study We used AD mice model to investigate the anti-AD activities of QF, as well as its underlying molecular mechanisms which involved signal transducer and activator of transcription 3 (STAT3), nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways. Materials and methods 2,4-dinitrofluorobenzene (DNFB)-induced AD mice were used to collect serum and skin tissues for consequential determination. The levels of various inflammatory factors [interleukin (IL)-12, Interferon (IFN)-γ, tumor necrosis factor (TNF)-α, IL-4, IL-6 and immunoglobulin E (IgE)] were determined by enzyme-linked immunosorbent assay (ELISA). Real-time polymerase chain reaction (RT-PCR) was contributed to detect the effects of relevant inflammatory factors on mRNA. The roles of STAT3, NF-κB and MAPK signaling pathways in AD response were analyzed by Western blotting (WB), and the thickening of mice dorsal skin and inflammatory cell infiltration were observed by hematoxylin and eosin (H&E) staining. Results QF significantly reduced the skin thickening, inflammatory cell infiltration and other symptoms in AD mice. The levels of IL-12, TNF-α, IL-4, IL-6 and IgE were decreased, while IFN-γ was increased by QF in the ELISA analysis. QF lessened the levels of lL-6 and elevated IFN-γ on the mRNA level. In addition, WB analysis showed QF thoroughly inhibited the activation of NF-κB, STAT3 and phosphorylation of JAK1, JAK2, JAK3, while partially suppressed MAPK signaling pathways. Conclusions QF inhibited the activations of STAT3, MAPK and NF-κB signaling pathways and possessed a significant therapeutic effect on AD. Therefore, QF deserves our continuous attention and research as a prominent medicine for AD.

On automatic block lines, there is a time interval between two trains traveling on a track in the same direction. Therefore, the dynamic load on the railway subgrade is composed of the periodic vibrations generated when the train passes, and the intermittent period when there is no train passing. To study the influence of the train interval time on the dynamic behavior of a railway subgrade, a series of dynamic triaxial tests of heavy-haul railway subgrade soil were conducted under consecutive cyclic loading and intermittent cyclic loading, while considering the interval time. The variation laws of the resilient moduli of specimens with different moisture contents and stress states with loading cycles under the two loading modes were compared and analyzed, and the influence of the load interval on the resilient modulus was discussed. The results showed that the load interval improved the resilient modulus of a specimen and its ability to resist failure. When the specimen was in a saturated state, its resilient modulus tended to increase with an increase in the intermittency period. A prediction model considering the soil stress state and load interval was proposed, based on introducing a factor k4 to quantify the effect of the load interval on the resilient modulus.

A novel gene encoding an endoglucanase designated Cel5D was cloned from a marine bacterium Martelella mediterranea by genomic library. The gene had a 1,113 bp opening reading frame encoding a 371-amino-acid protein with a molecular mass of 40,508 Da and containing a putative signal peptide (41 amino acids). Cel5D had low similarity (48-51% identity) with other known endoglucanases and consisted of one single catalytic domain, which belonged to the glycosyl hydrolase family 5. The maximum activity of Cel5D was observed at 60 degrees C and pH 5.0. Cel5D displayed broad pH stability within the range of pH 3.0-11.0 and retained hydrolytic activity in the presence of a wide variety of metal ions and some chemical reagents. These characteristics suggest that the enzyme has considerable potential in industrial applications.

Bacillus subtilis endo-beta-1,4-glucanase (Cel5A) hydrolyzes cellulose by cleavage of the internal bonds in the glucose chains, producing new ends randomly. Using directed evolution techniques of error-prone polymerase chain reaction (PCR) and DNA shuffling, several Cel5A variants with improved catalytic activity had been screened from the mutant library, which contained 71,000 colonies. Compared with the wild-type enzyme, the variants (M44-11, S75 and S78) showed 2.03 to 2.68-fold increased activities toward sodium carboxymethyl cellulose (CMC), while the M44-11 also exhibited a wider pH tolerance and higher thermostability. Structural models of M44-11, S75, S78, and WT proteins revealed that most of the substitutions were not located in the strictly conserved regions, except the mutation V255A of S75, which was closed to the nucleophile Glu257 in the catalytic center of the enzyme. Moreover, V74A and D272G of M44-11, which were not located in the substrate binding sites and the catalytic center, might result in improved stability and catalytic activity. These results provided useful references for directed evolution of the enzymes that belonged to the glycoside hydrolase family 5 (GH5).

Cancer cells amplify the expression of ASC amino acid transporter 2 (ASCT2, also called SLC1A5), a high-affinity glutamine carrier protein, to coordinate metabolic reprogramming and malignant transformation. Yet genetic and/or epigenetic mechanisms underlying the control of ASCT2-mediated glutamine metabolism remain to be clarified. Combined in-silico algorithms with systemic experimental screening, we herein identify the tumor suppressor miR-137 as an essential regulator that targets ASCT2 mRNA and cancer cell glutamine metabolism. Metabolic analysis shows that miR-137 derepression, similar to ASCT2 inactivation, significantly inhibits glutamine consumption and TCA cycle anaplerosis. Mechanistically, methyl-CpG binding protein 2 (MeCP2) and DNA methyltransferases (DNMTs) cooperate to promote active methylation of the miR-137 promoter and its decreased transcription, which conversely enhances ASCT2 expression and glutamine metabolism. As such, expression between miR-137 and ASCT2 is inversely correlated in multiple human cancer types, including colorectal carcinomas, glioblastomas, prostate and pancreatic cancers. These findings thus elucidate a universal mechanism responsible for ASCT2 deregulation in human cancers, revealing a molecular link between miR-137, ASCT2 and tumor metabolism. Citation Format: Xiao Daibiao, Junli Dong, Zihan Zhao, Ping Ren, Cong Li, Yufeng Hu, Jianguo Shi, Hexiu Su, Zhaojing Wang, Hudan Liu, Bo Li, Peng Gao, Guoliang Qing. Epigenetic silencing of microRNA-137 enhances ASCT2 expression and tumor glutamine metabolism [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1474. doi:10.1158/1538-7445.AM2017-1474

Conjugate gradient method is one of the most useful methods for solving unconstrained optimization problem. In this paper, we propose a hybrid conjugate gradient method for unconstrained optimization based on the Polak-Ribiere-Polyak and Dai-Yuan conjugate gradient Algorithms. By searching a particular direction, the new algorithm satisfies the sufficient descent condition naturally and satisfies the conjugacy condition. Furthemore under the Wolfe line search conditions, we prove that the new method can support the global convergence. The initial numerical experiments show that the new algorithm is efficient.

The anthrax toxin is an AB-type bacterium toxin composed of the protective antigen (PA) as the cell-binding B component, and the lethal factor (LF) and edema toxin (EF) as the catalytic A components. The PA component is a key factor in anthrax-related research and recombinant PA can be produced in general in Escherichia coli. However, such recombinant PA always forms inclusion bodies in the cytoplasm of E. coli, making difficult the procedure of its purification. In this study, we found that the solubility of recombinant PA was dramatically enhanced by fusion with glutathione S-transferase (GST) and an induction of its expression at 28 degrees C. The PA was purified to high homogeneity and a yield of 3 mg protein was obtained from 1 l culture by an affinity-chromatography approach. Moreover, we expressed and purified three PA mutants, I394C, A396C, and N435C, which were impaired in expression in previous study. Among them, a novel mutant N435C which conferred dominant-negative inhibitory activity on PA was identified. This new mutant may be useful in designing new antitoxin for anthrax prophylaxis and therapy.