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Abstract Many neurotransmitter receptors activate G proteins through exchange of GDP for GTP. The intermediate nucleotide-free state has eluded characterization, due largely to its inherent instability. Here we characterize a G protein variant associated with a rare neurological disorder in humans. Gαo K46E has a charge reversal that clashes with the phosphate groups of GDP and GTP. As anticipated, the purified protein binds poorly to guanine nucleotides yet retains wild-type affinity for G protein βγ subunits. In cells with physiological concentrations of nucleotide, Gαo K46E forms a stable complex with receptors and Gβγ, impeding effector activation. Further, we demonstrate that the mutant can be easily purified in complex with dopamine-bound D2 receptors, and use cryo-electron microscopy to determine the structure, including both domains of Gαo, without nucleotide or stabilizing nanobodies. These findings reveal the molecular basis for the first committed step of G protein activation, establish a mechanistic basis for a neurological disorder, provide a simplified strategy to determine receptor-G protein structures, and a method to detect high affinity agonist binding in cells.

BackgroundGrowing evidence emphasizes the importance of xanthophyll carotenoids and omega-3 fatty acids in eye health. However, the beneficial effects of such supplementation have not been thoroughly discussed among adults with high screen exposure. Current trial evidence on lutein bioavailability is contradictory, and the interactions of dietary intervention with host-related factors remain elusive. This study aims to investigate the comparative effectiveness of supplementation with macular xanthophylls and omega-3 fatty acids on macular pigment optical density (MPOD) and visual function, access the bioavailability of free lutein and lutein ester, and explore the complex interplay between genetic variations, intestinal microbiota, and the dietary intervention in Chinese adults with long-term exposure to digital devices.MethodsThe Lutein, Zeaxanthin, and Omega-3 (LZO) clinical trial is a 24-week multicenter, randomized, double-blind, placebo-controlled trial of 600 participants recruited from research centers, universities, and communities. Individuals are eligible to participate if they are aged over 18 years and use digital devices for over 8 h daily in the last 2 years, and will be randomized to six arms. A total of three visits will be scheduled at baseline, 12 and 24 weeks. The primary outcome is the change in MPOD over the 24-week intervention. The secondary outcomes are changes in visual function (visual acuity, best-corrected visual acuity, contrast and glare sensitivity, critical flicker fusion, reaction time, visuognosis persistence, symptoms and signs of dry eye, retinal thickness, and optical quality), and changes in serum lutein and zeaxanthin concentrations, and erythrocyte membrane omega-3 fatty acids. Genetic variations will be determined using genome-wide genotyping at baseline. 16S rRNA gene sequencing will be utilized to assess microbiome compositional changes before and after intervention.DiscussionThe trial is anticipated to establish early interventions to prevent photochemical ocular damage and delay the onset of vision impairment in young adults with long-term repeated exposure to screen-based electronic devices, and provide valuable insights for the development of precision nutrition strategies for maintaining eye health.Clinical trial registrationwww.clinicaltrials.in.th, Identifier, TCTR20220904002.

Cholesterol, an essential lipid of cell membranes, regulates G protein-gated inwardly rectifying potassium (GIRK) channel activity. Previous studies have shown that cholesterol activates GIRK2 homotetrameric channels, which are expressed in dopaminergic neurons of the brain. Deletion of GIRK2 channels affects both GIRK2 homo- and heterotetrames and can lead to abnormal neuronal excitability, including conditions such as epilepsy and addiction. A 3.5 Å cryo-EM structure of GIRK2 in complex with CHS (cholesteryl hemisuccinate) and PIP2 (phosphatidylinositol 4,5-bisphosphate) has been solved. This structure provides the opportunity to study GIRK2 channel gating dynamics regulated by cholesterol using gating molecular dynamics (GMD) simulations. In the present study, we conducted microsecond-long GMD simulations on the GIRK2 channel in its APO, PIP2, and PIP2/CHS bound states, followed by systematic analysis to gain molecular insights into how CHS modulates GIRK2 channel gating. We found that CHS binding facilitates GIRK2 channel opening, with 43 K+ ion permeation events observed, compared to 0 and 2 K+ ion permeation events for GIRK2-APO and GIRK2/PIP2, respectively. Binding of CHS to the GIRK2 channel enhances PIP2 and channel interactions, which is consistent with previous experimental results. The negatively charged PIP2 alters the internal electrostatic potential field in the channel and lowers the negative free energy barrier for K+ ion permeation.

With the expansion of the “Arctic Amplification” effect, the navigable period of the Arctic Northeast Passage (NEP) has gradually been lengthened, and its regular operation in summer has become a realistic possibility. Starting from the actual voyage data of Chinese merchant ships navigating the NEP, this paper analyzes the number of ships and voyages, ice class and ship type distribution, cargo volume and cargo type, transportation route distribution, navigable period and ice-breaking pilotage, and then calculates the profit of merchant ships. The results show that the voyage number and cargo volume of Chinese merchant ships shipping via the NEP are at a relatively low level, but in terms of the general trend, the number of voyages of Chinese merchant ships shipping via the NEP is increasing year by year, and the cargo volume of Chinese merchant ships is also increasing. With the accelerated melting of the Arctic sea ice, the navigable period for ships to pass has been prolonged, which has greatly increased the possibility of regular commercial operation in the NEP; At present, Chinese merchant ships have a chance to operate in the NEP; With the accelerated melting of sea ice, the period for Chinese merchant ships to pass has been extended, thus making it possible for the NEP to realize regular commercial operation.

Abstract The prevalence of porcine reproductive and respiratory syndrome virus 1 (PRRSV1) isolates has continued to increase in Chinese swine herds in recent years. However, no effective control strategy is available for PRRSV1 infection in China. In this study, we generated the first infectious cDNA clone (rHLJB1) of a Chinese PRRSV1 isolate and subsequently used it as a backbone to construct an ORF2-6 chimeric virus (ORF2-6-CON). This virus contained a synthesized consensus sequence of the PRRSV1 ORF2-6 gene encoding all the envelope proteins. The ORF2-6 consensus sequence shared > 90% nucleotide similarity with four representative strains (Amervac, BJEU06-1, HKEU16 and NMEU09-1) of PRRSV1 in China. ORF2-6-CON had replication efficacy similar to that of the backbone rHLJB1 virus in primary alveolar macrophages (PAMs) and exhibited cell tropism in Marc-145 cells. Piglet inoculation and challenge studies indicated that ORF2-6-CON is not pathogenic to piglets and can induce enhanced cross-protection against a heterologous SD1291 isolate. Notably, ORF2-6-CON inoculation induced higher levels of heterologous neutralizing antibodies (nAbs) against SD1291 than rHLJB1 inoculation, which was concurrent with a higher percentage of T follicular helper (Tfh) cells in tracheobronchial lymph nodes (TBLNs), providing the first clue that porcine Tfh cells are correlated with heterologous PRRSV nAb responses. The number of SD1291-strain-specific IFNγ-secreting cells was similar in ORF2-6-CON-inoculated and rHLJB1-inoculated pigs. Overall, our findings support that the Marc-145-adapted ORF2-6-CON can trigger Tfh cell and heterologous nAb responses to confer improved cross-protection and may serve as a candidate strain for the development of a cross-protective PRRSV1 vaccine.

Abstract The advance of genetic function indicators has enabled the observation of neuronal activities at single-cell resolutions. A major challenge for the applications on mammalian brains is the limited optical access depth. Currently, the method of choice to access deep brain structures is to insert miniature optical components. Among these validated miniature optics, the gradient-index (GRIN) lens has been widely employed for its compactness and simplicity. However, due to strong fourth-order astigmatism, GRIN lenses suffer from a small imaging field of view, which severely limits the measurement throughput and success rate. To overcome these challenges, we developed geometric transformation adaptive optics (GTAO), which enables adaptable achromatic large-volume correction through GRIN lenses. We demonstrate its major advances through in vivo structural and functional imaging of mouse brains. The results suggest that GTAO can serve as a versatile solution to enable large-volume recording of deep brain structures and activities through GRIN lenses.

Recent breakthroughs in the determination of atomic resolution 3-D cryo-electron microscopy structures of membrane proteins present an unprecedented opportunity for drug discovery. Structure-based drug discovery utilizing in silico methods enables the study of dynamic connectivity of stable conformations induced by the drug in achieving its effect. With the ever-expanding computational power, simulations of this type reveal protein dynamics in the nano-, micro-, and even millisecond time scales. In the present study, aiming to characterize the protein dynamics of the 5HT2A receptor stimulated by ligands (agonist/antagonist), we performed 1 µs MD simulations on 5HT2A/DOI (agonist), 5HT2A/GSK215083 (antagonist), and 5HT2A (APO, no ligand) systems. The crystal structure of 5HT2A/zotepine (antagonist) (PDB: 6A94) was used to set up the simulation systems in a lipid bilayer environment. We found the monitoring of the ionic lock residue pair (R3.50-E6.30) of 5HT2A in MD simulations to be a good approximation of the effects of agonists (ionic lock breakage) or antagonists (ionic lock formation) on receptor activation. We further performed analyses of the MD trajectories, including Principal Component Analysis (PCA), hydrogen bond, salt bridge, and hydrophobic interaction network analyses, and correlation between residues to identify key elements of receptor activation. Our results suggest that in order to trigger receptor activation, DOI must interact with 5HT2A through residues V5.39, G5.42, S5.43, and S5.46 on TM5, inducing significant conformational changes in the backbone angles of G5.42 and S5.43. DOI also interacted with residues W6.48 (toggle switch) and F6.51 on TM6, causing major conformational shifts in the backbone angles of F6.44 and V6.45. These structural changes were transmitted to the intracellular ends of TM5, TM6, and ICL3, resulting in the breaking of the ionic lock and subsequent G protein activation. The studies could be helpful in future design of selective agonists/antagonists for various serotonin receptors (5HT1A, 5HT2A, 5HT2B, 5HT2C, and 5HT7) involved in detrimental disorders, such as addiction and schizophrenia.

Introduction: The advent of lipid nanoparticles (LNPs) as a delivery platform for mRNA therapeutics has revolutionized the biomedical field, particularly in treating infectious diseases, cancer, genetic disorders, and metabolic diseases. Recent Advances in Therapeutic LNPs: LNPs, composed of ionizable lipids, phospholipids, cholesterol, and polyethylene glycol (PEG) lipids, facilitate efficient cellular uptake and cytosolic release of mRNA while mitigating degradation by nucleases. However, as synthetic entities, LNPs face challenges that alter their therapeutic efficacy and safety concerns. Toxicity/Reactogenicity/Immunogenicity: This review provides a comprehensive overview of the latest advancements in LNP research, focusing on preclinical safety assessments encompassing toxicity, reactogenicity, and immunogenicity. Summary and Outlook: Additionally, it outlines potential strategies for addressing these challenges and offers insights into future research directions for enhancing the application of LNPs in mRNA therapeutics.

Abstract Background Multimodal techniques-assisted resection of glioma under general anesthesia (GA) has been shown to achieve similar clinical outcomes as awake craniotomy (AC) in some studies. In this study, we aim to validate the use of multimodal techniques can achieve the maximal safe resection of high-grade glioma involving language areas (HGILAs) under GA. Methods HGILAs cases were reviewed and collected between January 2009 and December 2020 in our center. Patients were separated into multimodal group (using neuronavigation, intraoperative MRI combined with direct electrical stimulation [DES] and neuromonitoring [IONM]) and conventional group (neuronavigation alone) and clinical outcomes were compared between groups. Studies of HGILAs were reviewed systematically and the meta-analysis results of previous (GA or AC) studies were compared with our results. Results Finally, there were 263 patients in multimodal group and 137 patients in conventional group. Compared to the conventional group, the multimodal group achieved the higher median EOR (100% versus 94.32%, P

Abstract Objectives Advanced cancer patients face various symptoms, which can cause physical and psychological distress. As a multidimensional construct, spiritual well-being (SWB) may be an inner resource for dealing with these problems. Our study explored the impact of different dimensions of SWB on physical and psychological symptoms in advanced cancer patients admitted to a palliative care unit. Methods This cross-sectional study was conducted among 108 advanced cancer patients in the Hospice Ward, Shengjing Hospital of China Medical University. Patients completed questionnaires on SWB and cancer-related symptoms (insomnia, fatigue, pain, depression and anxiety) at the time of admission. Linear regression analysis was applied to determine the relationship between SWB (meaning, peace and faith) and symptom distress. Results SWB accounted for an additional variance of cancer-related symptoms (17.8% to 44.4%). Meaning was negatively associated with insomnia (β = -0.516, p < 0.001) and fatigue (β = -0.563, p < 0.001). Peace and faith were related to lower psychological symptoms, while meaning represented a positive effect on anxiety (β = 0.275, p = 0.036). Higher peace was associated with lower cancer pain (β = -0.422, p < 0.001). Conclusions Our findings suggested that achieving peace and faith appeared to function consistently as a positive resource for advanced cancer patients on depression, anxiety and pain, while meaning may serve to facilitate or hinder positive adjustment. Future studies should focus on the potential clinical implications by identifying the distinct dimension of SWB as symptom management targets in the palliative care practice.

Conclusion: DEX emerges as a promising option for palliative sedation in terminal-stage cancer patients. When used in conjunction with PCA, DEX has been shown to effectively, safely, and stably control refractory pain without inducing adverse effects such as respiratory/circulatory depression.

Abstract Background Although palliation of psycho-spiritual distress is of great importance in terminally ill cancer patients, there is a little information about screening patients who benefit from palliative care and identifying the cancer care targets. This study explored the relationship of pain management and positive expectations with depression, anxiety and spiritual well-being (SWB) in terminal cancer patients admitted to a palliative care unit. Methods Eighty-four terminal cancer inpatients were recruited from the Hospice Ward, Shengjing Hospital of China Medical University. Optimism and general self-efficacy (GSE) were evaluated at admission. Patients completed self-report questionnaires on SWB, depression, anxiety and pain both on admission and one week later. The repeated designed analysis of variance was used to explore the correlates of depression, anxiety and SWB (meaning, peace, faith). Results In our sample, only cancer pain diminished significantly one week later. For depression (p = 0.041) and faith (p = 0.013), there was a significant pain group (relieved vs. not relieved) × time interaction effect, such that those with satisfied pain control experienced the improved psycho-spiritual outcomes at 1 week. The relationship between positive expectations, peace and faith was also statistically significant, indicating that the improvement of peace or faith was significant in the low group of optimism and GSE. Conclusions Our findings indicated that pain management lied at the center of depression and SWB, meaning that effective pain management may reduce depression, and improve SWB among terminal cancer patients. Moreover, positive expectations, especially for optimism, may be the new target for SWB-related intervention research. Palliative care nurse should require the identification of terminal cancer patients who may more benefit from short-term palliative care, and target them with effective cancer care.

Abstract The procollagen C-protease enhancer (PCOLCE) has been identified to influence tumor growth and metastasis in multiple cancers. However, the relationship between PCOLCE activity and the progression of gliomas remains largely unknown. Glioma RNA-seq data were derived from the Chinese Glioma Genome Atlas (CGGA) and The Cancer Genome Atlas databases for analysis. Kaplan–Meier survival curve, clinical characterization correlation, univariate and multivariate Cox, and receiver operating characteristic curve analyses were performed to assess the prognostic role of PCOLCE. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and Gene Set Enrichment Analysis were used to determine the functions or pathways associated with PCOLCE. The ESTIMATE and CIBERSORT algorithms, Spearman’s rank correlation analysis, and Tumor Immune Estimation Resource (TIMER) databases were used to explore the relationship between PCOLCE and immune infiltration. Correlation analysis between PCOLCE, related genes, and immune cell markers was conducted using the TIMER database. Immunophenoscore assays were performed to determine differential PCOLCE expression levels in glioma. The sensitivity of multi-drugs were determined to explore potential chemotherapeutic agents in between PCOLCE. Compared to normal brain tissue, PCOLCE expression was increased in glioma and correlated with shorter overall survival (OS). Furthermore, significant differences were observed in the immune scores and immune cell infiltration levels. PCOLCE is positively associated with immune checkpoints and many immune markers. Additionally, PCOLCE expression was higher in gliomas with higher IPS Z-scores in CGGA. High expression of PCOLCE increased sensitivity to multiple chemotherapy agents in CGGA (P

Link failures are the most common type of fault in software-defined networking (SDN), which is an extremely crucial aspect of SDN fault tolerance. Existing strategies include proactive and reactive approaches. Proactive schemes pre-deploy backup paths for fast recovery but may exhaust resources, while reactive schemes calculate paths upon failure, resulting in longer recovery but better outcomes. This paper proposes a single link failure recovery strategy that combines these two schemes, termed as flow-aware pro-reactive (FAPR), with the aim of achieving high-speed recovery while ensuring high-quality backup paths. Specifically, the controller adopts pro-VLAN to install backup paths for each link into switches, and precalculates multiple backup paths for each link in the controller before any link failures. In case of a link failure, pro-VLAN, i.e., a method based on the proactive approach, is initially utilized for swift recovery automatically without the involvement of the controller. Simultaneously, the controller analyzes types of affected flows based on the transport layer data, obtains several key network indicators of the backup paths, and then selects the most suitable path for different flows on the basis of the current network view. Simulation results and theoretical analysis show that the recovery time of the FAPR scheme reduces by over 65% compared with the reactive scheme. The interruption rate of flows after fault recovery is reduced by 20% and 50% compared with the reactive and proactive schemes, respectively. In addition, due to the principle of pro-VLAN, the number of backup flow rules required is at most 85% less than that required by the proactive scheme. In conclusion, FAPR promises the highest failure recovery speed and the lowest interruption rate among three methods, and helps to improve the quality of network services.

Abstract The role of excision repair cross-complementation group 6-like (ERCC6L) has been reported in several cancers, but little is known about its expression and function in laryngeal squamous cell carcinoma (LSCC). In this study, the expression of ERCC6L in LSCC was determined by immunohistochemistry and its correlation with prognostic factors was analyzed. Furthermore, cytological functional validation elucidated the role and underlying mechanisms of ERCC6L dysregulation in LSCC. Our data revealed that ERCC6L expression was elevated in LSCC and it’s correlated with TNM stage. In addition, ERCC6L knockdown LSCC cells showed decreased proliferation and migration, increased apoptosis, and reactive oxygen species (ROS). Mechanically, overexpression of ERCC6L promoted nuclear translocation of FOXM1 to facilitate direct binding to the KIF4A promoter and upregulated KIF4A expression. Furthermore, KIF4A knockdown attenuated the role of ERCC6L overexpression in promoting proliferation, migration, and tumorigenesis of LSCC cells. In summary, ERCC6L promoted the binding of FOXM1 and KIF4A in LSCC cells to drive their progression, which may be a promising target for precision therapy in this disease.

The authors develop an optical gearbox to accelerate the laser scanning microscopes, achieving flexible adjustment of imaging frame rate from tens of Hz to 1 kHz. The technology is validated through in vivo functional imaging of mice brains.

Ni interlayer was applied in the lap welding of Ti6Al4V titanium (Ti) alloy and 301 stainless steel (SS). By adjusting the welding parameters, the SS irradiated by the laser was not completely penetrated. The SS/Ni and Ni/Ti interfaces were metallurgically bonded due to indirect heating. Two bonding interfaces formed based on solid phase diffusion bonding and eutectic reaction. As the laser power increased, the thickness of the remaining Ni interlayer gradually decreased. A Ni transition layer, as barrier to mixing of the two base materials can prevent the formation of Ti–Fe IMCs. The shear test results showed that the shear strength of the joint increased first and then decreased with the increase of laser power, and the maximum shear strength of joint was 269.8 MPa.

Parthenolide (PTL) is a new compound extracted from traditional Chinese medicine. In recent years, it has been proven to play an undeniable role in tumors, autoimmune diseases, and inflammatory diseases. Similarly, an increasing number of experiments have also confirmed the biological mechanism of PTL in these diseases. In order to better understand the development trend and potential hot spots of PTL in cancer and other diseases, we conducted a detailed bibliometric analysis. The purpose of presenting this bibliometric analysis was to highlight and inform researchers of the important research directions, co-occurrence relationships and research status in this field. Publications related to PTL research from 2002 to 2022 were extracted on the web of science core collection (WoSCC) platform. CiteSpace, VOSviewers and R package “bibliometrix” were applied to build relevant network diagrams. The bibliometric analysis was presented in terms of performance analysis (including publication statistics, top publishing countries, top publishing institutions, publishing journals and co-cited journals, authors and co-cited authors, co-cited references statistics, citation bursts statistics, keyword statistics and trend topic statistics) and science mapping (including citations by country, citations by institution, citations by journal, citations by author, co-citation analysis, and keyword co-occurrence). The detailed discussion of the results explained the focus and latest trends from the bibliometric analysis. Finally, the current status and shortcomings of the research field on PTLwere clearly pointed out for reference by scholars.

Copper-based nanomaterials have been widely used in catalysis, electrodes, and other applications due to their unique electron-transfer properties. In this work, an efficient electrochemical sensor based on an electrode modified with one-dimensional Cu(OH)2/carboxymethyl cellulose (CMC) composite nanofibers was fabricated and investigated for the detection of aspirin. Scanning electron microscopy was employed to examine the morphological characteristics of these composite nanofibers. Cyclic voltammetry and electrochemical impedance spectroscopy were used to assess the electrochemical performance of a Cu(OH)2/CMC composite nanofiber-modified electrode. The findings indicate that the modified electrode has a very high sensitivity to aspirin. The observed enhanced performance could be a result of the high surface-to-volume ratio of the composite nanofibers and their superior electron-transport characteristics, which may hasten electron transfer between aspirin and the surfaces of the modified electrode. This detection technique also demonstrated strong selectivity for aspirin. These findings imply that the technique can be applied as a highly effective and selective approach to aspirin measurement in biological science.

Piezoelectric ultrasonic transducers have shown great potential in biomedical applications due to their high acoustic-to-electric conversion efficiency and large power capacity. The focusing technique enables the transducer to produce an extremely narrow beam, greatly improving the resolution and sensitivity. In this work, we summarize the fundamental properties and biological effects of the ultrasound field, aiming to establish a correlation between device design and application. Focusing techniques for piezoelectric transducers are highlighted, including material selection and fabrication methods, which determine the final performance of piezoelectric transducers. Numerous examples, from ultrasound imaging, neuromodulation, tumor ablation to ultrasonic wireless energy transfer, are summarized to highlight the great promise of biomedical applications. Finally, the challenges and opportunities of focused ultrasound transducers are presented. The aim of this review is to bridge the gap between focused ultrasound systems and biomedical applications.

Flexible capacitive pressure sensors as one of the crucial sensing components have aroused widespread concern because of their vital role in development of wearable artificial devices, healthcare biomonitoring and human-machine interface. However, there has been a considerable challenge to successfully fabricate them with a tradeoff between high sensitivity and wide sensing range by low-cost and convenient manufacturing methods. Herein, the orientated thermoplastic polyurethane elastomer rubber nanofiber membranes (TPU-O NMs) were prepared via conjugated electrospinning to build a dielectric layer for the flexible capacitive wide-range pressure sensors with high sensitivity. In addition, polyaniline (PANI) was in situ polymerized on the TPU-O NMs as flexible electrodes to maintain the conductive pathway, flexibility, and breathability of the sensors. The TPU@PANI NMs form a bamboo-raft-like microstructure between the aligned fibers under pressure, resulting in a significant increase in sensitivity and working range. Furthermore, the conductive TPU@PANI NMs can be stretched to 1600% without fracture. The assembled sensor has a high sensitivity of 31.73 kPa−1 and a fast response/recovery time of 96 ms. Meantime, it also exhibits a minimum detection limit of 1 Pa, an excellent stability of 10,000 cycles, and a good linearity in a broad working age (1 Pa∼122.5 kPa). Finally, the capacitive pressure sensor was successfully applied for full-range detection of human motions, such as finger bending, joint bending, and even boxing movements. Hence this highly sensitive and wide-range pressure sensor can be a promising candidate for healthcare monitoring, motion recognition, and wearable circuitry in artificial intelligence.

Non-small cell lung cancer (NSCLC) is one of the most serious diseases affecting human health today, and current research is focusing on gut flora. There is a correlation between intestinal flora imbalance and lung cancer, but the specific mechanism is not clear. Based on the “lung and large intestine being interior-exteriorly related” and the “lung-intestinal axis” theory. Here, based on the theoretical comparisons of Chinese and western medicine, we summarized the regulation of intestinal flora in NSCLC by active ingredients of traditional Chinese medicine and Chinese herbal compounds and their intervention effects, which is conducive to providing new strategies and ideas for clinical prevention and treatment of NSCLC.

ABSTRACT Pathogen inactivation is a strategy to improve the safety of transfusion products. The only pathogen reduction technology for blood products currently approved in the US utilizes a psoralen compound, called amotosalen, in combination with UVA light to inactivate bacteria, viruses, and protozoa. Psoralens have structural similarity to bacterial multidrug efflux pump substrates. As these efflux pumps are often overexpressed in multidrug-resistant pathogens, we tested whether contemporary drug-resistant pathogens might show resistance to amotosalen and other psoralens based on multidrug efflux mechanisms through genetic, biophysical, and molecular modeling analysis. The main efflux systems in Enterobacterales, Acinetobacter baumannii, and Pseudomonas aeruginosa are tripartite resistance-nodulation-cell division (RND) systems, which span the inner and outer membranes of Gram-negative pathogens, and expel antibiotics from the bacterial cytoplasm into the extracellular space. We provide evidence that amotosalen is an efflux substrate for the E. coli AcrAB, Acinetobacter baumannii AdeABC, and P. aeruginosa MexXY RND efflux pumps. Furthermore, we show that the MICs for contemporary Gram-negative bacterial isolates for these species and others in vitro approached and exceeded the concentration of amotosalen used in the approved platelet and plasma inactivation procedures. These findings suggest that otherwise safe and effective inactivation methods should be further studied to identify possible gaps in their ability to inactivate contemporary, multidrug-resistant bacterial pathogens. IMPORTANCE Pathogen inactivation is a strategy to enhance the safety of transfused blood products. We identify the compound, amotosalen, widely used for pathogen inactivation, as a bacterial multidrug efflux substrate. Specifically, experiments suggest that amotosalen is pumped out of bacteria by major efflux pumps in E. coli, Acinetobacter baumannii, and Pseudomonas aeruginosa. Such efflux pumps are often overexpressed in multidrug-resistant pathogens. Importantly, the MICs for contemporary multidrug-resistant Enterobacterales, Acinetobacter baumannii, Pseudomonas aeruginosa, Burkholderia spp., and Stenotrophomonas maltophilia isolates approached or exceeded the amotosalen concentration used in approved platelet and plasma inactivation procedures, potentially as a result of efflux pump activity. Although there are important differences in methodology between our experiments and blood product pathogen inactivation, these findings suggest that otherwise safe and effective inactivation methods should be further studied to identify possible gaps in their ability to inactivate contemporary, multidrug-resistant bacterial pathogens.

IntroductionSugar-sweetened beverage (SSB) intake is associated with an increased risk of cardiometabolic diseases. However, evidence regarding associations of artificially sweetened beverages (ASBs) and fruit juices with cardiometabolic diseases is mixed. In this study, we aimed to investigate the association between the SSB, ASB and fruit juice consumption with the incidence of cardiometabolic conditions and mortality.MethodsRelevant prospective studies were identified by searching PubMed, Web of Science, Embase, and Cochrane Library until December 2022 without language restrictions. The pooled relative risk (RR) and 95% confidence intervals (CIs) were estimated for the association of SSBs, ASBs, and fruit juices with the risk of type 2 diabetes (T2D), cardiovascular disease (CVD), and mortality by using random-effect models.ResultsA total of 72 articles were included in this meta-analysis study. Significantly positive associations were observed between the consumption of individual beverages and T2D risk (RR: 1.27; 95% CI: 1.17, 1.38 for SSBs; RR: 1.32; 95% CI: 1.11, 1.56 for ASBs; and RR:0.98; 95% CI: 0.93, 1.03 for fruit juices). Moreover, our findings showed that intakes of SSBs and ASBs were significantly associated with risk of hypertension, stroke, and all-cause mortality (RR ranging from 1.08 to 1.54; all p

Automated ICD coding via machine learning that focuses on some specific diseases has been a hot topic. As one of the leading causes of death, coronary heart diseases (CHD) have seldom been specifically studied by related research, probably due to lack of data concretely targeting at the diseases. Based on Fuwai-CHD and MIMIC–III–CHD, which are a private dataset from Fuwai Hospital and the CHD-related subset of a public dataset named MIMIC-III respectively, this study aimed at automated CHD coding by a deep learning method, which mainly consists of three modules. The first is a BERT variant module responsible for encoding clinical text. In the module, we fine-tuned BERT variants with masked language model on clinical text, and proposed a truncation method to tackle the problem that BERT variants generally cannot handle sequences containing more than 512 tokens. The second is a word2vec module for encoding code titles and the third is a label-attention module for integrating the embeddings of clinical text and code titles. In short, we named the method BW_att. We compared BW_att against some widely studied baselines, and found that BW_att performed best in most of the coding missions. Specifically, BW_att reached a Macro-F1 of 96.2% and a Macro-AUC of 98.9% for the top-100 most frequent codes in Fuwai-CHD, which covered 89.2% of the total code occurrences. When predicting the top-50 most frequent codes in MIMIC–III–CHD, BW_att reached a Macro-F1 of 40.5% and a Macro-AUC of 66.1%. Moreover, BW_att was capable of locating informative tokens from clinical text for predicting the target codes. In summary, BW_att can not only suggest CHD codes accurately, but also possess robust interpretability, hence has great potential in facilitating CHD coding in practice.

Boolean function in symmetric cryptographic algorithm must satisfy excellent cryptographic criteria to ensure the security of the algorithm.By combining the advantages of tabu search algorithm and hill climbing algorithm, a new heuristic generation algorithm called hybrid tabu search algorithm for excellent Boolean function was proposed.A large number of Boolean function with high nonlinearity, low autocorrelation, one-resilient, optimal algebraic degree, optimal algebraic immunity, optimal (suboptimal) resistance to fast algebraic attacks could be obtained quickly by applying the proposed algorithm.Simulation results demonstrate that the cryptographic properties of the Boolean function obtained by the proposed algorithm with strong search ability and fast running speed are better than the results of known optimization algorithm.Moreover, the algorithm also provides good Boolean function that cannot be obtained by using construction method.

Multi-objective optimal dispatching schemes with intelligent algorithms are recognized as effective measures to promote the economics and environmental friendliness of microgrid applications. However, the low accuracy and poor convergence of these algorithms have been challenging for system operators. The bird swarm algorithm (BSA), a new bio- heuristic cluster intelligent algorithm, can potentially address these challenges; however, its computational iterative process may fall into a local optimum and result in premature convergence when optimizing small portions of multi-extremum functions. To analyze the impact of a multi-objective economic–environmental dispatching of a microgrid and overcome the aforementioned problems of the BSA, a self-adaptive levy flight strategy-based BSA (LF–BSA) was proposed. It can solve the dispatching problems of microgrid and enhance its dispatching convergence accuracy, stability, and speed, thereby improving its optimization performance. Six typical test functions were used to compare the LF–BSA with three commonly accepted algorithms to verify its excellence. Finally, a typical summer-time daily microgrid scenario under grid-connected operational conditions was simulated. The results proved the feasibility of the proposed LF–BSA, effectiveness of the multi- objective optimization, and necessity of using renewable energy and energy storage in microgrid dispatching optimization.

Abstract Background Parthenolide has anti-inflammatory, immunomodulatory and anti-cancer activities. But its effect on thyroid cancer cells is still largely unknown. Methods Label-free quantitative proteomics and bioinformatics analysis were used to investigate the differentially expressed proteins and their functions in thyroid cancer treated with parthenolide and control pair. Hoechst 33258 fluorescent staining and Annexin V-FITC/PI double staining flow cytometry were used to detected BCPAP cells apoptosis. Parallel reaction monitoring (PRM) and quantitative real-time PCR were used to verify the expression of apoptosis-related differential proteins and their mRNA. Results Sixty up-regulated and 96 down-regulated differentially expressed proteins were identified in parthenolide treated thyroid cancer cells BCPAP compared with control thyroid cancer cells. The proteins were mainly relevant to various biological processes that included metabolic processes, response to extracellular stimulus and interaction with host. The molecular functions of most differentially expressed proteins were associated with binding functions and nucleotidyltransferase activity. According to the Kyoto Encyclopedia of Genes and Genomes, the differentially expressed proteins identified are primarily related to various types of metabolic pathways and DNA replication. In cell experiments in vitro, with the increase of the dose of parthenolide, the number of cells gradually decreased, the apoptosis rate gradually increased. PRM verified that the apoptosis-related proteins HMOX1 and GCLM were up-regulated and IL1B was down-regulated in BCPAP cells treated with parthenolide. The mRNA expressions of HMOX1, GCLM, ITGA6 and CASP8 were up-regulated and HSPA1A was down-regulated by PCR. Conclusions Parthenolide may influence the biological behavior of human thyroid cancer cells by affecting the expression of proteins related to cell metabolism and DNA replication. Parthenolide induced significant cellular morphological changes and apoptosis in human thyroid cancer cells, leading to an anti-proliferative effect.

ObjectivesConsidering the high-level requirements of cooperative design ability, along with the fact that the working interface between general arrangement and aesthetic design of luxury cruise ships is unclear to a certain degree, this paper focuses on the constraints of general arrangement on the aesthetic design.MethodBased on regional deconstruction and an exploration of the parameter tree of influencing factors on the appearance design of luxury cruise ships, the main factors are selected, constraints in ship code conventions, ship safety and engineering design experience are analyzed, and the boundary conditions for the creativity of appearance design are obtained.ResultsIn terms of appearance and shape features such as the overall outline, visual envelope of the side view, and funnel, functional zones at the bow and the stern of ship, the constraint conditions of appearance design are confirmed through the relative dimensions of different parameters to avoid obstructing the free flow of creative work.ConclusionsThe results of this study can provide significant references for exploring the cooperative mode between engineering design and appearance design, and improving the level of domestic innovative cruise ship design.

Saccharin is a commonly used artificial sweetener that exhibits both sweetening and sweet inhibition activities. The species‐dependent response towards saccharin and the interaction between saccharin and the sweet taste receptor T1R2/T1R3 remain elusive. In this study we used mismatched chimeras of T1R2 and T1R3 and calcium mobilization functional analysis to reveal a detailed species‐dependent response towards saccharin of human, squirrel monkey, and mouse sweet taste receptors. Our findings, combined with previous results by others, suggest multiple and complex interaction modes between saccharin and the sweet taste receptor, which are helpful guidelines for effective modulation of the sweet taste by sweetener/sweet inhibitors.

Abstract Diffuse and multi-lobes involved glioma (DMG) is a rare disease, and the aim of this study was to assess the role of multimodal-assisted surgical resection of tumours combined with chemoradiotherapy and identify prognosis. Clinical data were collected from 38 patients with a diagnosis of DMG. Nineteen patients received multimodal-assisted surgical resection of tumours combined with chemoradiotherapy, and another 19 patients underwent chemoradiotherapy alone after stereotactic puncture biopsy. The clinical characteristics, magnetic resonance imaging (MRI) findings, histopathological diagnosis, progression-free survival, and overall survival of DMG patients were retrospectively analysed. Twenty-six males and 12 females were included, and the age of the participants ranged from 10 to 80 years (46.34 ± 15.61). The median overall survival in our study was 25 months, and the progression-free survival was 17 months. The extent of resection was 50.10–73.60% (62.54% ± 7.92%). The preoperative and the postoperative KPS score of the patients in the operation group showed no statistically significant difference. The results of logistic regression demonstrated that overall survival was positively associated with operative treatment + chemoradiotherapy (p = 0.003) but negatively associated with age and corpus callosal involvement (p = 0.028 and 0.022, respectively). Kaplan–Meier analyses showed that those who underwent surgical treatment had a significant progression-free and overall survival benefit compared to those who did not undergo surgical treatment (log-rank test; p = 0.011 and 0.008, respectively). Older age and involvement of the corpus callosum represent a poor prognosis in DMG patients. Multimodal-assisted surgical resection of tumours combined with chemoradiotherapy might be a treatment option for DMG. Further research is needed to obtain the clear evidence of the effect of surgical treatment.

Neural modulation plays a major role in delineating the circuit mechanisms and serves as the cornerstone of neural interface technologies. Among the various modulation mechanisms, ultrasound enables noninvasive label-free deep access to mammalian brain tissue. To date, most if not all ultrasonic neural modulation implementations are based on ∼1 MHz carrier frequency. The long acoustic wavelength results in a spatially coarse modulation zone, often spanning over multiple function regions. The modulation of one function region is inevitably linked with the modulation of its neighboring regions. Moreover, the lack of in vivo cellular resolution cell-type-specific recording capabilities in most studies prevents the revealing of the genuine cellular response to ultrasound. To significantly increase the spatial resolution, we explored the application of high-frequency ultrasound. To investigate the neuronal response at cellular resolutions, we developed a dual-modality system combining in vivo two-photon calcium imaging and focused ultrasound modulation. The studies show that the ∼30 MHz ultrasound can suppress the neuronal activity in awake mice at 100-μm scale spatial resolutions, paving the way for high-resolution ultrasonic neural modulation. The dual-modality in vivo system validated through this study will serve as a general platform for studying the dynamics of various cell types in response to ultrasound.

ABSTRACT The Escherichia coli acriflavine resistance protein D (AcrD) is an efflux pump that belongs to the resistance-nodulation-cell division (RND) superfamily. Its primary function is to provide resistance to aminoglycoside-based drugs by actively extruding these noxious compounds out of E. coli cells. AcrD can also mediate resistance to a limited range of other amphiphilic agents, including bile acids, novobiocin, and fusidic acids. As there is no structural information available for any aminoglycoside-specific RND pump, here we describe cryo-electron microscopy (cryo-EM) structures of AcrD in the absence and presence of bound gentamicin. These structures provide new information about the RND superfamily of efflux pumps, specifically, that three negatively charged residues central to the aminoglycoside-binding site are located within the ceiling of the central cavity of the AcrD trimer. Thus, it is likely that AcrD is capable of picking up aminoglycosides via this central cavity. Through the combination of cryo-EM structural determination, mutagenesis analysis, and molecular simulation, we show that charged residues are critically important for this pump to shuttle drugs directly from the central cavity to the funnel of the AcrD trimer for extrusion. IMPORTANCE Here, we report cryo-EM structures of the AcrD aminoglycoside efflux pump in the absence and presence of bound gentamicin, posing the possibility that this pump is capable of capturing aminoglycosides from the central cavity of the AcrD trimer. The results indicate that AcrD utilizes charged residues to bind and export drugs, mediating resistance to these antibiotics.

Sweet taste is a primary sensation for the preference and adaption of primates to diet, which is crucial for their survival and fitness. It is clear now that the sweet perception is mediated by a G protein-coupled receptor (GPCR)-sweet taste receptor T1R2/T1R3, and many behavioral or physiological experiments have described the diverse sweet taste sensitivities in primates. However, the structure-function relationship of T1R2s/T1R3s in primates, especially the molecular basis for their species-dependent sweet taste, has not been well understood until now. In this study, we performed a comprehensive sequence, structural and functional analysis of sweet taste receptors in primates to elucidate the molecular determinants mediating their species-dependent sweet taste recognition. Our results reveal distinct taxonomic distribution and significant characteristics (interaction, coevolution and epistasis) of specific key function-related residues, which could partly account for the previously reported behavioral results of taste perception in primates. Moreover, the prosimians Lemuriformes species, which were reported to have no sensitivity to aspartame, could be proposed to be aspartame tasters based on the present analysis. Collectively, our study provides new insights and promotes a better understanding for the diversity, function and evolution of sweet taste receptors in primates.

PurposeTo establish a predictive model to predict the occurrence of language deficit for patients after surgery of glioma involving language areas (GILAs) under general anesthesia (GA).MethodsPatients with GILAs were retrospectively collected in our center between January 2009 and December 2020. Clinical variables (age, sex, aphasia quotient [AQ], seizures and KPS), tumor-related variables (recurrent tumor or not, volume, language cortices invaded or not, shortest distance to language areas [SDLA], supplementary motor area or premotor area [SMA/PMA] involved or not and WHO grade) and intraoperative multimodal techniques (used or not) were analyzed by univariate and multivariate analysis to identify their association with temporary or permanent language deficits (TLD/PLD). The predictive model was established according to the identified significant variables. Receiver operating characteristic (ROC) curve was used to assess the accuracy of the predictive model.ResultsAmong 530 patients with GILAs, 498 patients and 441 patients were eligible to assess TLD and PLD respectively. The multimodal group had the higher EOR and rate of GTR than conventional group. The incidence of PLD was 13.4% in multimodal group, which was much lower than that (27.6%, P

Abstract Extra-virgin olive oil (EVOO) is a critical component of the Mediterranean diet, which has been found beneficial to human health. Bitterness is often positively associated with the presence of phenolic compounds in EVOO. There are twenty-five bitter taste receptors (TAS2Rs) in humans, each of which responds to specific bitter tastants. The identity of phenolic compounds and the bitter taste receptors they stimulate remain unknown. In this study, we isolated 12 phenolic and secoiridoid compounds from the olive fruit and the oil extracted from it, and tested their ability to stimulate bitter taste receptor activity, using a calcium mobilization functional assay. Our results showed that seven out of twelve studied compounds activated TAS2R8, and five of them activated TAS2R1, TAS2R8, and TAS2R14. The phenolic compounds oleuropein aglycon and ligstroside aglycon were the most potent bitter tastants in olive oil. TAS2R1 and TAS2R8 were the major bitter taste receptors activated most potently by these phenolic compounds. The results obtained here could be utilized to predict and control the bitterness of olive oil based on the concentration of specific bitter phenolics produced during the milling process of olives.

Abstract The objectives of the study were to comprehend the prevalence of hypertension (HTN) and prehypertension (PHT), awareness, treatment, and control of HTN and its distribution in urban, agricultural, pastoral, and semi‐agricultural/semi‐pastoral areas, and to explore the related factors of HTN among Mongolian adults in China. From August 2018 to August 2020, a multi‐stage stratified cluster random sampling method was conducted to investigate the prevalence of HTN among Mongolian adults aged ≥18 years living in China (n = 2558). Inclusion criteria for HTN were systolic blood pressure ≥ 140 mm Hg and/or diastolic blood pressure ≥ 90 mm Hg and/or had hypertensive history and/or taking antihypertensive drugs for HTN. The prevalence rates of HTN and PHT were 44.77% and 32.03%, respectively. The prevalence rates of PHT in urban, agricultural, pastoral, and semi‐agricultural/semi‐pastoral areas were 34.93%, 34.73%, 26.03%, and 33.44%, respectively, and the prevalence rates of HTN were 35.97%, 40.15%, 49.68%, and 48.07%, respectively. The awareness, treatment and control rates of HTN were 66.48%, 58.93%, and 16.48%, respectively. In this survey, the overweight, obesity, and central obesity rates were 34.30%, 30.67%, and 58.08%, respectively. Compared with Chinese adults ≥18 years, the prevalence rate of HTN among Mongolian adults in China aged ≥ 18 years was relatively high; the prevalence rate of PHT and HTN awareness, treatment, and control rates were similar. The prevalence of HTN and the rates of obesity and central obesity were higher in pastoral regions than in the other three types of regions, and the rate of overweight was highest in agricultural regions.

A Ti/Al joint with an Nb transition layer was obtained, where aluminum alloy and Nb sheet partially melted to form the weld and diffusion bonding was formed at Ti/Nb interface. The remaining transition layer Nb acted as a diffusion barrier between TC4 titanium alloy and 6082 aluminum alloy thus successfully prevented the formation of brittle TiAl, TiAl2 and TIAl3 phases. A flat Ti–Nb diffusion zone composed of (Ti, Nb) solid solution was formed at Ti/Nb interface, while a narrow Al–Nb reaction zone was formed at the Nb/Al–FZ (aluminum alloy fusion zone) interface with the main microstructures of NbAl3, Nb2Al and Al-based solid solution. The simulation of welding temperature field and thermal cycle curve by finite element software was conducted to provide theoretical data support for clarifying the formation of the joint and interface reaction mechanism. The mechanical property of the joint was determined by the Al–Nb reaction zone which could be witnessed by observing the fracture location and hardness distribution of the joint. The joint fractured at the Al–Nb reaction zone and adjacent Al–FZ with the maximum tensile strength of 288.97 MPa.

Abstract Background Hyperextension bicondylar tibial plateau fracture (HBTPF) is a particular form of tibial plateau fracture which has gained increasing interest recently but were rarely documented. In this study, we reported the characteristics, clinical intervention, and therapeutic outcomes of HBTPF patients. Methods From May 2015 to October 2017, clinical data of consecutive patients with bicondylar tibial plateau fractures (BTPF) who underwent surgical treatment in our hospital were retrospectively studied. The patients were allocated to either the HBTPF group (study group) or the non-HBTPF group (control group) based on the radiological features, and inclusion and exclusion criteria. Demographics, characteristics of knee joint injuries, complications, and outcomes were compared between the two groups. Results In total, 59 patients were included in this study. Among them, 17 patients with HBTPF were identified and 42 patients were diagnosed as non-HBTPF. No differences in age, sex, cause of injury, side of injury, site of injury, nerve injury, operation time, and treatment time and incision complication between HBTPF and non-HBTPF group. The incidence rate of popliteal artery injury in HBTPF group was 29.4 %, which was significantly higher than that of non-HBTPF group. Small bone chips on the lateral film were found in 94.1 % of the patients in HBTPF group, which was significantly higher than that of non-HBTPF group. The range of motion (ROM) and hospital for special surgery (HSS) score of HBTPF group were significantly lower than those of non-HBTPF group. Conclusions HBTPF is a severe injury with a higher incidence rate of popliteal artery injury and worse outcomes than non-HBTPF. Small bone chips at the anterior margin of the proximal tibia on the lateral plain film might be a characteristic of HBTPF.

In this study, a model for the development of the wide floodplain in the lower Yellow River Basin, in China was developed. This model includes flood control schemes using grading criteria, enables sediment deposition in partitioned zones, and allows free exchange between channel runoff and sediment. The wide floodplain located between the main channel and levees is divided into three typical regions: the tender, low, and high floodplains. Different ecological models should be applied when these floodplains are constructed. This paper describes the associated research ideas and methodology, and clarifies several key issues, including sediment prediction and regulation, land planning, land use, and a multi-dimensional framework of safeguard measures for industries on the lower Yellow River floodplain. A refined ecological development model is proposed for the lower Yellow River floodplain, and future work on ecological and sustainable development of the lower floodplain is suggested. To establish a comprehensive system integrating runoff and sediment resource regulation in the Yellow River Basin, future work should focus on runoff and sediment exchange mechanisms in the wandering lower reaches. Furthermore, it is necessary to improve theories on floodplain planning and ecological construction, and these theories should be integrated with the research findings on land development across the lower Yellow River floodplain.

Abstract Background Our goal was to clarify the comparison between elective neck dissection (END) and the wait-and-see policy in neck management for cT1N0 buccal squamous cell carcinoma (SCC). Methods This was a retrospective comparison of 175 prospectively enrolled patients with cT1N0 buccal SCC. The patients were divided into two groups based on the nonrandomized management of the neck: 125 patients received END, and 50 patients were exposed to the wait-and-see policy. The main study endpoints were locoregional control (LRC) and disease-specific survival (DSS). Patients were asked to complete the shoulder domain in the University of Washington quality of life questionnaire, version 4, 1 year postoperatively. Results Ten of the patients undergoing END developed recurrence, and the 5-year LRC rate was 92%. Five patients undergoing the wait-and-see policy developed recurrence, and the 5-year LRC rate was 90%. The difference was not significant (p = 0.668). There were 6 deaths in patients undergoing END, and the 5-year DSS rate was 94%. There were 3 deaths in patients undergoing the wait-and-see policy, and the 5-year DSS rate was 94%; the difference was not significant (p = 0.777). The mean shoulder scores of patients undergoing END and the wait-and-see policy were 93.9 and 100, respectively, and the difference was not significant (p = 0.284). Conclusion Elective neck dissection does not carry a survival benefit compared to the wait-and-see policy, and it is not suggested for patients with cT1N0 buccal SCC.

Wingless/Int-1 (Wnt) signaling is one of the most well-known oncogenic pathways. Numerous studies have uncovered an aberrant expression of Wnt in cancer and its association with multiple oncogenic processes, such as cell proliferation, epithelial–mesenchymal transition (EMT), and invasiveness. Most previous studies mainly focused on the canonical branch of Wnt signaling pathway, i.e., Wnt/β-catenin signaling. The Wnt/planar cell polarity (PCP) signaling pathway, as the most recently described branch of Wnt signaling, was much less investigated in oncology research. In this study, we thoroughly characterized the activity of the Wnt/PCP regulation pathway in low-grade glioma (LGG) patients. Subtyping based on the expression pattern of the Wnt/PCP regulation pathway revealed three (C1–C3) subgroups with significant survival differences. Each group displayed distinct genomic characteristics. For instance, C1 was enriched with capicua transcriptional repressor (CIC) truncating mutations and 1p19q codel. C2 was characterized with tumor protein p53 (TP53) and ATRX chromatin remodeler (ATRX) inactivating mutations but depletion of telomerase reverse transcriptase (TERT) promoter mutations. C3 showed elevated malignancy reflected from several oncogenic characteristics, such as tumor heterogeneity and cell stemness, and demonstrated the worst survival outcome. In addition, C3 showed elevated macrophage segregation via induction of cytokines that are able to enhance the permeability of the brain–blood barrier (BBB). Lastly, we developed a prognostic model based on the risk score system. Validation indicated that our model can independently predict the prognosis of LGG patients.

G protein-gated inwardly rectifying K+ (GIRK) channels form highly active heterotetramers in the body, such as in neurons (GIRK1/GIRK2 or GIRK1/2) and heart (GIRK1/GIRK4 or GIRK1/4). Based on three-dimensional atomic resolution structures for GIRK2 homotetramers, we built heterotetrameric GIRK1/2 and GIRK1/4 models in a lipid bilayer environment. By employing a urea-based activator ML297 and its molecular switch, the inhibitor GAT1587, we captured channel gating transitions and K+ ion permeation in sub-microsecond molecular dynamics (MD) simulations. This allowed us to monitor the dynamics of the two channel gates (one transmembrane and one cytosolic) as well as their control by the required phosphatidylinositol bis 4-5-phosphate (PIP2). By comparing differences in the two trajectories, we identify three hydrophobic residues in the transmembrane domain 1 (TM1) of GIRK1, namely, F87, Y91, and W95, which form a hydrophobic wire induced by ML297 and de-induced by GAT1587 to orchestrate channel gating. This includes bending of the TM2 and alignment of a dipole of two acidic GIRK1 residues (E141 and D173) in the permeation pathway to facilitate K+ ion conduction. Moreover, the TM movements drive the movement of the Slide Helix relative to TM1 to adjust interactions of the CD-loop that controls the gating of the cytosolic gate. The simulations reveal that a key basic residue that coordinates PIP2 to stabilize the pre-open and open states of the transmembrane gate flips in the inhibited state to form a direct salt-bridge interaction with the cytosolic gate and destabilize its open state.

The mycobacterial membrane protein large 3 (MmpL3) transporter is essential and required for shuttling the lipid trehalose monomycolate (TMM), a precursor of mycolic acid (MA)-containing trehalose dimycolate (TDM) and mycolyl arabinogalactan peptidoglycan (mAGP), in Mycobacterium species, including Mycobacterium tuberculosis and Mycobacterium smegmatis. However, the mechanism that MmpL3 uses to facilitate the transport of fatty acids and lipidic elements to the mycobacterial cell wall remains elusive. Here, we report 7 structures of the M. smegmatis MmpL3 transporter in its unbound state and in complex with trehalose 6-decanoate (T6D) or TMM using single-particle cryo-electron microscopy (cryo-EM) and X-ray crystallography. Combined with calculated results from molecular dynamics (MD) and target MD simulations, we reveal a lipid transport mechanism that involves a coupled movement of the periplasmic domain and transmembrane helices of the MmpL3 transporter that facilitates the shuttling of lipids to the mycobacterial cell wall.