Dissecting the Role of the Hydroxyl Moiety at C14 in (+)-Opioid-Based TLR4 Antagonists via Wet-Lab Experiments and Molecular Dynamics Simulations.

Toll-like receptor 4 (TLR4) is pivotal as an innate immune receptor, playing a critical role in mediating neuropathic pain and drug addiction through its regulation of the neuroinflammatory response. The nonclassical (+)-opioid isomers represent a unique subset of TLR4 antagonists known for their effective blood-brain barrier permeability. Despite growing interest in the structure-activity relationship of these (+)-opioid-based TLR4 antagonists, the specific impact of heteroatoms on their TLR4 antagonistic activities has not been fully explored. This study investigated the influence of the hydroxyl group at C14 in six (+)-opioid TLR4 antagonists (1-6) using wet-lab experiments and in silico simulations. The corresponding C14-deoxy derivatives (7-12) were synthesized, and upon comparison with their corresponding counterparts (1-6), it was discovered that their TLR4 antagonistic activities were significantly diminished. Molecular dynamics simulations showed that the (+)-opioid TLR4 antagonists (1-6) possessed more negative binding free energies to the TLR4 coreceptor MD2, which was responsible for ligand recognition. This was primarily attributed to the formation of a hydrogen bond between the hydroxyl group at the C-14 position of the antagonists (1-6) and the R90 residue of MD2 during the binding process. Such an interaction facilitated the entry and subsequent binding of these molecules within the MD2 cavity. In contrast, the C14-deoxy derivatives (7-12), lacking the hydroxyl group at the C-14 position, missed this crucial hydrogen bond interaction with the R90 residue of MD2, leading to their egression from the MD2 cavity during simulations. This study underscores the significant role of the C14 hydroxyl moiety in enhancing the effectiveness of (+)-opioid TLR4 antagonists, which provides insightful guidance for designing future (+)-isomer opioid-derived TLR4 antagonists.

Exploring the Function of (+)-Naltrexone Precursors: Their Activity as TLR4 Antagonists and Potential in Treating Morphine Addiction.

Disruptions in the toll-like receptor 4 (TLR4) signaling pathway are linked to chronic inflammation, neuropathic pain, and drug addiction. (+)-Naltrexone, an opioid-derived TLR4 antagonist with a (+)-isomer configuration, does not interact with classical opioid receptors and has moderate blood-brain barrier permeability. Herein, we developed a concise 10-step synthesis for (+)-naltrexone and explored its precursors, (+)-14-hydroxycodeinone (1) and (+)-14-hydroxymorphinone (3). These precursors exhibited TLR4 antagonistic activities 100 times stronger than (+)-naltrexone, particularly inhibiting the TLR4-TRIF pathway. In vivo studies showed that these precursors effectively reduced behavioral effects of morphine, like sensitization and conditioned place preference by suppressing microglial activation and TNF-α expression in the medial prefrontal cortex and ventral tegmental area. Additionally, 3 displayed a longer half-life and higher oral bioavailability than 1. Overall, this research optimized (+)-naltrexone synthesis and identified its precursors as potent TLR4 antagonists, offering potential treatments for morphine addiction.

Configurable circular-polarization-dependent optoelectronic silent state for ultrahigh light ellipticity discrimination.

Filterless light-ellipticity-sensitive optoelectronic response generally has low discrimination, thus severely hindering the development of monolithic polarization detectors. Here, we achieve a breakthrough based on a configurable circular-polarization-dependent optoelectronic silent state created by the superposition of two photoresponses with enantiomerically opposite ellipticity dependences. The zero photocurrent and the significantly suppressed noise of the optoelectronic silent state singularly enhance the circular polarization extinction ratio (CPER) and the sensitivity to light ellipticity perturbation. The CPER of our device approaches infinity by the traditional definition. The newly established CPER taking noise into account is 3-4 orders of magnitude higher than those of ordinary integrated circular polarization detectors, and it remains high in an expanded wavelength range. The noise equivalent light ellipticity difference goes below 0.009° Hz-1/2 at modulation frequencies above 1000 Hz by a light power of 281 µW. This scheme brings a leap in developing monolithic ultracompact circular polarization detectors.

Graphene/MoS2-xOx/graphene photomemristor with tunable non-volatile responsivities for neuromorphic vision processing.

Conventional artificial intelligence (AI) machine vision technology, based on the von Neumann architecture, uses separate sensing, computing, and storage units to process huge amounts of vision data generated in sensory terminals. The frequent movement of redundant data between sensors, processors and memory, however, results in high-power consumption and latency. A more efficient approach is to offload some of the memory and computational tasks to sensor elements that can perceive and process the optical signal simultaneously. Here, we proposed a non-volatile photomemristor, in which the reconfigurable responsivity can be modulated by the charge and/or photon flux through it and further stored in the device. The non-volatile photomemristor has a simple two-terminal architecture, in which photoexcited carriers and oxygen-related ions are coupled, leading to a displaced and pinched hysteresis in the current-voltage characteristics. For the first time, non-volatile photomemristors implement computationally complete logic with photoresponse-stateful operations, for which the same photomemristor serves as both a logic gate and memory, using photoresponse as a physical state variable instead of light, voltage and memresistance. The polarity reversal of photomemristors shows great potential for in-memory sensing and computing with feature extraction and image recognition for neuromorphic vision.

Geometry-asymmetric photodetectors from metal-semiconductor-metal van der Waals heterostructures.

The functional diversities of two-dimensional (2D) material devices with simple architectures are ultimately limited by immature doping techniques. An alternative strategy is to use geometry-asymmetric metal-semiconductor-metal (GA-MSM) structures, which enable the basic functions of semiconductor junctions such as rectification and photovoltaics. Here, the mixed-dimensional van der Waals heterostructures (MDvdWHs) based on the separation and self-assembly of p-type SnS layered nanosheets (NSs) and n-type SnS2 nanoparticles (NPs) are obtained using an aqueous phase exfoliation (APE) method. Due to the surface charge transfer doping, the carrier transport mechanism of devices based on MDvdWHs turns from thermionic field emission (TFE) to thermionic emission (TE), with the rectification factor (Iforward/Ireverse) changing from 0.7 to 3. To further illustrate the experimental results, the generic current transport models of GA-MSM devices have been established based on the TE and TFE mechanisms in which the TE and TFE mechanisms lead to opposite rectification phenomena in good agreement with the experimental results. The GA-MSM devices show a photovoltaic effect with a high responsivity of 35 A W-1 and detectivity of 3.4 × 1011 cm Hz1/2 W-1. This study not only provides a novel strategy to design photovoltaic devices with MDvdWHs, but more importantly, we have established fundamental models for the rectification behavior of GA-MSM devices.

HDAC4 depletion ameliorates IL-13-triggered inflammatory response and mucus production in nasal epithelial cells via activation of SIRT1/NF-κB signaling.

INTRODUCTION: Allergic rhinitis (AR) is frequently known as a chronic respiratory disease with a global high prevalence. The pivotal roles of histone deacetylase 4 (HDAC4) in multiple human diseases have been underlined by numerous studies. Nevertheless, whether HDAC4 is implicated in AR remains to be elaborated. The objective of the current study is to clarify the impacts of HDAC4 on AR. METHODS: First, human nasal epithelial cells (hNECs) were pretreated by interleukin-13 (IL-13). HDAC4 expression in hNECs with the presence or absence of IL-13 treatment was tested by quantitative reverse-transcription polymerase chain reaction (RT-qPCR) and western blot. Following, after HDAC4 was depleted, levels of histamine, Immunoglobulin E (IgE) and inflammatory factors were analyzed by ELISA assay. Then, Mucin-5AC (MUC5AC) expression was examined through RT-qPCR, western blot, and IF assay. Western blot was to analyze the expression of sirtuin 1 (SIRT1)/nuclear factor-kappaB (NF-κB) signaling-related proteins. After IL-13-induced hNECs were cotransfected with HDAC4 interference plasmids and SIRT1 inhibitor EX527, the functional experiments above were conducted again. RESULTS: The experimental data in this study presented that HDAC4 expression was increased in IL-13-induced hNECs. Silencing of HDAC4 cut down the levels of histamine, IgE and inflammatory factors and the expression of MUC5AC. Additionally, knockdown of HDAC4 led to the activation of SIRT1/NF-κB signaling. Further, the downregulated levels of histamine, IgE and inflammatory factors and the expression of MUC5AC imposed by HDAC4 interference were all reversed by EX527. CONCLUSIONS: In short, HDAC4 inhibition activated SIRT1/NF-κB signaling to mitigate inflammatory response and mucus production in IL-13-treated nasal epithelial cells in AR.

Pristine PN junction toward atomic layer devices.

In semiconductor manufacturing, PN junction is formed by introducing dopants to activate neighboring electron and hole conductance. To avoid structural distortion and failure, it generally requires the foreign dopants localize in the designated micro-areas. This, however, is challenging due to an inevitable interdiffusion process. Here we report a brand-new junction architecture, called "layer PN junction", that might break through such limit and help redefine the semiconductor device architecture. Different from all existing semiconductors, we find that a variety of van der Waals materials are doping themselves from n- to p-type conductance with an increasing/decreasing layer-number. It means the capability of constructing homogeneous PN junctions in monolayers' dimension/precision, with record high rectification-ratio (>105) and low cut-off current (<1 pA). More importantly, it spawns intriguing functionalities, like gate-switchable-rectification and noise-signal decoupled avalanching. Findings disclosed here might open up a path to develop novel nanodevice applications, where the geometrical size becomes the only critical factor in tuning charge-carrier distribution and thus functionality.

2D materials-based homogeneous transistor-memory architecture for neuromorphic hardware.

In neuromorphic hardware, peripheral circuits and memories based on heterogeneous devices are generally physically separated. Thus, exploration of homogeneous devices for these components is key for improving module integration and resistance matching. Inspired by the ferroelectric proximity effect on two-dimensional (2D) materials, we present a tungsten diselenide­on­lithium niobate cascaded architecture as a basic device that functions as a nonlinear transistor, assisting the design of operational amplifiers for analog signal processing (ASP). This device also functions as a nonvolatile memory cell, achieving memory operating (MO) functionality. On the basis of this homogeneous architecture, we also investigated an ASP-MO integrated system for binary classification and the design of ternary content-addressable memory for potential use in neuromorphic hardware.

Multi-stage responsive peptide nanosensor: Anchoring EMT and mitochondria with enhanced fluorescence and boosting tumor apoptosis.

Epithelial-mesenchymal transition (EMT) is closely related to tumor metastasis and invasion. Thereinto, mesenchymal tumor mitochondria are the critical target for tumor inhibition. Therefore, real-time in vivo monitoring of EMT as well as inhibiting mesenchymal tumor mitochondria is of great diagnosis and therapy significance. Herein, we construct a multi-stage recognition and morphological transformable self-assembly-peptide nano biosensor NDRP which can response the EMT marker and specifically damage the mesenchymal tumor cell in vivo. This nano-molar-affinity sensor is designed and screened with sensitive peptides containing a molecular switching which could be specifically triggered by the receptor to achieve the vesicle-to-fibril transformation in living system with enhanced fluorescent signal. NDRP nanosensor could target the tumor lesion in circulatory system, recognize mesenchymal tumor marker DDR2 (Discoidin domain receptor 2) in cellular level and specifically achieve mitochondria in subcellular level as well as damaged mitochondria which could be applied as a in vivo theranostic platform.

Interferon characterization associates with asthma and is a potential biomarker of predictive diagnosis.

Interferon (IFN) plays a role in immune and inflammation responses. However, the effect of IFN in asthma is still not fully clear. The present study was conducted to better understand the role of IFN signatures in asthma. Blood samples from case-control studies (study 1: 348 asthmas and 39 normal controls and validation study 2: 411 asthmas and 87 normal controls) were enrolled. The single-sample gene set enrichment analysis (ssGSEA) method was used to quantify the levels of 74 IFN signatures. Gene Ontology analysis and pathway function analysis were performed for functional analysis and a protein-protein interaction (PPI) network was constructed. The area under the curve (AUC) value was used to evaluate the diagnostic ability. In our work, IFN-γ response-DN, negative regulation of IFN-γ secretion, IFNG pathway, negative regulation of response to IFN-γ, and type 1 IFN biosynthetic process showed higher levels in asthma. Functional analysis demonstrated that pathway and biological process involved in IFN signaling pathway, regulation of type 1 IFN production and response to IFN-γ. Hub IFN-related genes were identified, and their combination as biomarker exhibited a good diagnostic capacity for asthma (AUC = 0.832). These findings offered more insight into the underlying mechanism of how IFN signatures affected asthma. The use of the easy-to-apply IFN-related genes might serve as a promising blood-based biomarker for early diagnosis of asthma.

Effect of Huangqin Tang on Colonic Gene Expression in Rats with Ulcerative Colitis.

In this study, we explored the pharmacological mechanisms of Huangqin Tang (HQT; a traditional Chinese medicine formula) in ulcerative colitis (UC) and provided evidence for potential roles HQT plays by gene expression profiling. The UC rat model was made via a compound method (trinitrobenzene sulfonic acid plus ethanol). After a ten-day treatment, microarray analysis was performed from the colon segment of the rats. Biological functions and specific signaling pathways were enriched based on differentially expressed genes (DEG), and corresponding gene networks were constructed via Ingenuity Pathway Analysis (IPA). Through the network, we screened the potential "candidate targets," such as ITGB1, FN1, CASP3, and ITGA5 and FABP1, ABCB1, FABP2, and SLC51B. These potential candidate targets were functionally related to immune responses, inflammation, and metabolism. Moreover, HQT significantly decreased serum levels of proinflammatory factors nitrogen monoxide (NO), proinflammatory cytokines interleukin- (IL-) 17, and prostaglandin E2 (PGE2). The degree of HE staining of colonic tissue was severe in the model group but reduced significantly in the HQT group. HQT exhibited protective effects against colon damage by inhibiting the inflammatory response.

Overexpression of hypoxia-inducible factor 1α is associated with neutrophilic inflammation in chronic rhinosinusitis with nasal polyps.

OBJECTIVE: This study aimed to assess the possible role of hypoxia-inducible factor 1α (HIF-1α) in promoting neutrophilic inflammation in chronic rhinosinusitis with nasal polyps (CRSwNP) patients. METHODS: We examined HIF-1α expression in sinonasal tissues from CRSwNP patients and healthy controls by using immunohistochemistry, qRT-PCR, and western blot. Next, the stimulatory effects of several cytokines (IFN-γ, IL-17A, IL-6, etc.) and reagents (dexamethasone (DEX), clarithromycin (CAM) and curcumin (CUM)) on HIF-1α expression in cultured normal nasal epithelial cells (NECs) were also evaluated. Moreover, the effects of CAM and glucocorticoid on nasal symptoms and signs of uncontrolled neutrophilic CRSwNP patients were evaluated. RESULTS: The mRNA and protein expression of HIF-1α were significantly increased in polyp tissues compared with healthy controls (P < 0.05), and the HIF-1α level in polyp tissues was positively associated with IL-17A production and tissue neutrophilia (P < 0.05). Moreover, in cultured NECs, HIF-1α expression was upregulated in the presence of IL-17A and IL-6 (P < 0.05). Both CAM and CUM showed an additive effect with DEX in inhibiting HIF-1α expression (P < 0.05). Moreover, combined glucocorticoid and CAM significantly improved nasal symptoms and signs compared with glucocorticoid alone in uncontrolled neutrophilic CRSwNP patients (P < 0.05). CONCLUSION: Our findings indicate that HIF-1α is associated with neutrophilic inflammation and glucocorticoid resistance in CRSwNP patients.

Inhibition of TLR4 inhibits allergic responses in murine allergic rhinitis by regulating the NF-κB pathway.

The present study investigated the underlying mechanisms and effects of toll-like receptor 4 (TLR4) on a mouse model of allergic rhinitis (AR). An ovalbumin (OVA)-induced mouse model of AR was treated with TLR4-short hairpin RNA (shRNA). Allergic symptoms were then subsequently assessed. Protein levels of OVA-specific immunoglobulin E (IgE), eosinophil cation protein (ECP), leukotriene C4 (LTC4) and prostaglandin D2 (PGD2) in mice serum and nasal lavage fluid, as well as various inflammatory cytokine mediators in mice serum, were determined by ELISA. Protein level detection was performed using reverse transcription-quantitative PCR and western blot analysis. The results revealed that TLR4 was highly expressed in the nasal mucosa of AR mice. TLR4 inhibition significantly relieved OVA-induced AR symptoms. Relief of symptoms was evidenced by a decreased frequency of sneezing and nose friction, reduced levels of OVA-specific IgE, ECP, LTC4, PGD2, less inflammatory cells and decreased levels of T-helper 2 type cytokines. In addition, the data indicated that OVA-induced activation of the NF-κB pathway was repressed by TLR4-shRNA. The results of the current study indicate that TLR4 may be a promising therapeutic target of AR.

[Effect of Huangqin Tang on the function of regulatory TLR4/MyD88 signal pathway in rats with ulcerative colitis].

This study was designed to investigate the effect of Huangqin Tang (HQT) on TLR4/Myd88 pathway and the downstream cytokines in rats with ulcerative colitis (UC) to explore its underlying mechanisms of action. The model of UC rats with cell immunoreactivity was made using a compound method (trinitrobenzene sulfonic acid plus ethanol). Rats were randomly divided into the control group, the model group, the salazosulfapyridine (SASP) group, high, medium and low dose(20, 10, 5 g·kg-1) of HQT groups. After a three-day treatment, production of NO in serum was detected by Griess assay, the levels of interleukin (IL)-4, IL-10, IL-17 and prostaglandin E2（PGE2） in serum were detected by ELISA. After a five-day treatment, the positive protein expressions of COX-2 and iNOS in the colon tissue were determined by ICH method, the protein expressions of TLR4 and MyD88 in colon tissue were determined by Western blot. Compared with the control group, the levels of NO, IL-17, PGE2, the protein expressions of TLR4, MyD88 and the protein positive expressions of COX-2, iNOS were apparently higher in the model group. Compared with model group, the above indexes were significantly improved in the SASP and high-dose HQT groups (P < 0.05). These results show that HQT has a definite effect on UC in rats. Its mechanisms of action may be achieved by inhibiting the activity of TLR4/MyD88/NF-κB signal pathway and down-regulation of NO, IL-17 and PGE2 production.

Targeted exome sequencing reveals novel USH2A mutations in Chinese patients with simplex Usher syndrome.

BACKGROUND: Usher syndrome (USH) is an autosomal recessive disorder characterized by hearing impairment and vision dysfunction due to retinitis pigmentosa. Phenotypic and genetic heterogeneities of this disease make it impractical to obtain a genetic diagnosis by conventional Sanger sequencing. METHODS: In this study, we applied a next-generation sequencing approach to detect genetic abnormalities in patients with USH. Two unrelated Chinese families were recruited, consisting of two USH afflicted patients and four unaffected relatives. We selected 199 genes related to inherited retinal diseases as targets for deep exome sequencing. Through systematic data analysis using an established bioinformatics pipeline, all variants that passed filter criteria were validated by Sanger sequencing and co-segregation analysis. RESULTS: A homozygous frameshift mutation (c.4382delA, p.T1462Lfs*2) was revealed in exon20 of gene USH2A in the F1 family. Two compound heterozygous mutations, IVS47 + 1G > A and c.13156A > T (p.I4386F), located in intron 48 and exon 63 respectively, of USH2A, were identified as causative mutations for the F2 family. Of note, the missense mutation c.13156A > T has not been reported so far. CONCLUSION: In conclusion, targeted exome sequencing precisely and rapidly identified the genetic defects in two Chinese USH families and this technique can be applied as a routine examination for these disorders with significant clinical and genetic heterogeneity.