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Ultrasound (US) washing has been verified to improve the quality of postharvest blueberry fruit. However, its physiological and molecular mechanisms remain largely unknown. In the present study, an US with a frequency of 25 kHz and a power density of 400 W for 2 min was performed to investigate its role in impacting the quality of blueberries. The results showed that US washing improved the quality of blueberries, with a higher firmness and lignin content (p < 0.05) than the control. Moreover, US washing inhibited the levels of superoxide radical (O2·−) production rate and hydrogen peroxide (H2O2) content while stimulating the superoxide dismutase (SOD) and catalase (CAT) activities of the blueberry fruit. Transcriptomic analysis screened 163 differentially expressed genes (DEGs), and the key DEGs were mainly enriched in phenylpropanoid biosynthesis, flavonoid biosynthesis, and plant–pathogen interaction pathways. Furthermore, the transcription factors and the structural genes associated with lignin biosynthesis were also identified from the DEGs. More importantly, the correlation analysis revealed that firmness and lignin content were positively correlated with the expression of C4H, COMT1, and POD52 in blueberry fruit, indicating that these genes might be involved in the regulation of US-mediated lignin synthesis. The findings provide new insight into the US-enhanced quality of blueberry fruits.

Cutibacterium granulosum, a commensal bacterium found on human skin, formerly known as Propionibacterium granulosum, rarely causes infections and is generally considered non-pathogenic. Recent research has revealed the transferability of the multidrug-resistant plasmid pTZC1 between C. granulosum and Cutibacterium acnes, the latter being an opportunistic pathogen in surgical site infections. However, there is a noticeable lack of research on the genome of C. granulosum, and the genetic landscape of this species remains largely uncharted. We investigated the genomic features and evolutionary structure of C. granulosum by analyzing a total of 30 Metagenome-Assembled Genomes (MAGs) and isolate genomes retrieved from public databases, as well as those generated in this study. A pan-genome of 6,077 genes was identified for C. granulosum. Remarkably, the ‘cloud genes’ constituted 62.38% of the pan-genome. Genes associated with mobilome: prophages, transposons [X], defense mechanisms [V] and replication, recombination and repair [L] were enriched in the cloud genome. Phylogenomic analysis revealed two distinct mono-clades, highlighting the genomic diversity of C. granulosum. The genomic diversity was further confirmed by the distribution of Average Nucleotide Identity (ANI) values. The functional profiles analysis of C. granulosum unveiled a wide range of potential Antibiotic Resistance Genes (ARGs) and virulence factors, suggesting its potential tolerance to various environmental challenges. Subtype I-E of the CRISPR-Cas system was the most abundant in these genomes, a feature also detected in C. acnes genomes. Given the widespread distribution of C. granulosum strains within skin microbiome, our findings make a substantial contribution to our broader understanding of the genetic diversity, which may open new avenues for investigating the mechanisms and treatment of conditions such as acne vulgaris.

Summary: Rabies viral vectors have become important components of the systems neuroscience toolkit, allowing both direct retrograde targeting of projection neurons and monosynaptic tracing of inputs to defined postsynaptic populations, but the rapid cytotoxicity of first-generation (ΔG) vectors limits their use to short-term experiments. We recently introduced second-generation, double-deletion-mutant (ΔGL) rabies viral vectors, showing that they efficiently retrogradely infect projection neurons and express recombinases effectively but with little to no detectable toxicity; more recently, we have shown that ΔGL viruses can be used for monosynaptic tracing with far lower cytotoxicity than the first-generation system. Here, we introduce third-generation (ΔL) rabies viral vectors, which appear to be as nontoxic as second-generation ones but have the major advantage of growing to much higher titers, resulting in significantly increased numbers of retrogradely labeled neurons in vivo. Motivation: Rabies viral vectors are useful tools for labeling projection neurons, but first-generation vectors are cytotoxic, and second-generation vectors are difficult to produce at high concentrations, and this limits the numbers of labeled neurons. Here, we introduce third-generation vectors that are both nontoxic and easily grown to high concentrations, allowing labeling of many more neurons in vivo.

Introduction: Biomimetic lubricant-infused porous surfaces are developed and applied for omniphobicity and corrosion protection, which exhibit great advantages compared to superhydrophobic surfaces.Methods: Herein, superhydrophobic Fe@E-Zn@PFOA was prepared via the electrodeposition of laminated Zinc coating, further vapor etching, and post-modification with perfluoro caprylic acid. The facile, inexpensive, and environment-friendly water vapor etching process can form a porous honeycomb-like structure. Moreover, the perfluoropolyether lubricant was wicked into the porous and superhydrophobic surfaces, obtaining lubricant-infused surfaces of Fe@E-Zn@PFOA@PFPE.Results and discussion: The influences of the textured roughness and chemical composition on the surface wettability were systematically investigated. The Fe@E-Zn@PFOA@PFPE performs omniphobicity with small sliding angles and superior corrosion resistance compared with the superhydrophobic surface, owing to their multiple barriers, including infused lubricant, hydrophobic monolayers, and compact Zn electroplating coating. Thus, the proposed lubricant-infused surface may provide insights into constructing protective coatings for the potential applications of engineering metal materials.

Abstract Purpose The purpose of this study was to observe the harm of circadian misalignment (CM), caused by an inverted photoperiod (IP), on the hearts of the adolescent Wistar rats, and to explore the mechanisms leading to harm. Methods An IP was used to create a CM model. A total of 174 Wistar rats were randomly divided into circadian alignment (CA) and CM groups (87 rats per group). The different activity rhythms of the two groups of rats were adjusted through different light/dark cycles for 90 days. We recorded the rhythmic activity trajectory and sleep time of the rats. After 90 days of modeling, we performed various analyses (i.e., blood pressure, weight, cardiac ultrasound tests, serological tests, cardiac tissue immunofluorescence, immunohistochemistry, transmission electron microscopy on myocardial mitochondria, western blotting, and quantitative polymerase chain reactions). Results (1) The IP protocol caused CM in rats. (2) CM rats showed significantly higher blood pressure during the day (resting phase). They also showed significantly higher serum levels of angiotensin II and epinephrine during the day compared to the CA rats. (3) CM caused up-regulation of gene expression of adrenergic receptors α1 (α1-AR) and β1 (β1-AR) and down-regulation of the glucocorticoid receptor (Gr) gene expression in rat hearts. It also caused downregulation of Bmal1 expression. In addition, the changes in Bmal1 and Per2 correlated with the changes in β1-AR and α1-AR. (4) CM had adverse effects on multiple molecular proteins of the heart. (5) CM increased the collagen fibers in the rat heart and increased the destruction of mitochondria. (6) Eventually, CM caused a decrease in the pumping function of the heart and decreased the coronary blood flow rate. Conclusions (1) CM significantly affected the cardiac structure and function in the adolescent rats through a variety of mechanisms. (2) CM can regulate the expression of myocardial clock genes, and it is likely to have an impact on the heart through this pathway.

ABSTRACT: In this study, the effects of naringin on hepatic yolk precursors formation and antioxidant capacity of Three-Yellow breeder hens during late laying period were evaluated. A total of 480 (54-wk-old) Three-Yellow breeder hens were randomly assigned to 4 groups (6 replicates of 20 hens): nonsupplemented control diet (C), and control diet supplemented with 0.1%, 0.2%, and 0.4% of naringin (N1, N2, and N3), respectively. Results showed that dietary supplemented with 0.1%, 0.2%, and 0.4% of naringin for 8 wk promoted the cell proliferation and attenuated the excessive fat accumulation in the liver. Compared with C group, increased concentrations of triglyceride (TG), total cholesterol (T-CHO), high-density lipoprotein cholesterol (HDL-C), and very low-density lipoprotein (VLDL), and decreased contents of low-density lipoprotein cholesterol (LDL-C) were detected in liver, serum and ovarian tissues (P < 0.05). After 8 wk of feeding with naringin (0.1%, 0.2%, and 0.4%), serum estrogen (E2) level, expression levels of proteins and genes of estrogen receptors (ERs) increased significantly (P < 0.05). Meanwhile, naringin treatment regulated expression of genes related to yolk precursors formation (P < 0.05). Furthermore, dietary naringin addition increased the antioxidants, decreased the oxidation products, and up-regulated transcription levels of antioxidant genes in liver tissues (P < 0.05). These results indicated that dietary supplemented with naringin could improve hepatic yolk precursors formation and hepatic antioxidant capacity of Three-Yellow breeder hens during the late laying period. Doses of 0.2% and 0.4% are more effective than dose of 0.1%.

The potential toxicity of microplastic (MPs) to organisms has attracted extensive attention. However, due to the subacute toxicity of MPs, the biological effect is hard to verify in short-term exposure experiment. Here, by tracking the dynamics of gut microbes, mice model was utilized to evaluate the toxicity of compositional MPs (PE, PET, PP, PS and PVC). After 7 days digestive exposure, the physiological indicators were normal as the control group that the body weight and serum cholesterol levels were insignificant change. Whereas, through histopathological examination, all the treatment groups suffered colon tissue damage, among which PS had the most inflammatory cells. Moreover, the high-throughput sequencing results revealed great variation of intestinal flora in treated mice. The ratio of Bacteroidetes and Firmicutes in PE, PET and PP treatment groups heighten, and the relative abundance of Ruminococcaceae and Lachnospiraceae increased significantly at family levels. At the genus level, Alistipes bacteria in PS treatment group significantly decreased that is associated with obesity risk. It indicated that MPs induced inflammatory response would further interfere the dynamics of intestinal flora causing health effect in living organisms. This work shed light on MPs toxicity in short-term exposure and supplied research paradigm of MPs health risk assessment.

Abstract In this paper, we first consider the stability problem for a class of stochastic quaternion-valued neural networks with time-varying delays. Next, we cannot explicitly decompose the quaternion-valued systems into equivalent real-valued systems; by using Lyapunov functional and stochastic analysis techniques, we can obtain sufficient conditions for mean-square exponential input-to-state stability of the quaternion-valued stochastic neural networks. Our results are completely new. Finally, a numerical example is given to illustrate the feasibility of our results.

ABSTRACT: In this study, the effects of 3 graded dietary levels (0.1%, 0.2%, and 0.4%) of naringin were studied in Three-Yellow breeder hens during the late laying period (55–62 wk). A total of 480 Three-Yellow breeder hens (54-wk-old) were randomly divided into 4 groups (6 replicates of 20 hens): basal diet group (C), and basal diets supplemented with 0.1%, 0.2%, and 0.4% of naringin (N1, N2, and N3), respectively. Results showed that dietary supplementation with 0.1%, 0.2%, and 0.4% of naringin for 8 wk increased the laying rate and egg mass, enhanced egg yolk color, and decreased the feed egg ratio (P < 0.05). Meanwhile, compared with hens in C group, there were more preovulatory follicles and higher ovarian index as well as an enhanced ovarian somatic cell proliferation in hens of N2 and N3 groups (P < 0.05). With 0.2% and 0.4% naringin, glutathione concentration, the activity of catalase and total superoxide dismutase, and the total antioxidant capacity of ovarian tissues and serum increased (P < 0.05), while the contents of malondialdehyde and hydrogen peroxide decreased (P < 0.05). Moreover, compared to C group, the transcription levels of antioxidant genes in ovarian tissues increased in hens from N2 and N3 groups (P < 0.05). In conclusion, supplementation with 0.2% and 0.4% naringin both could improve the laying rate, ovarian and serum antioxidant capacity of Three-Yellow breeder hens during the late laying period.

The current standard treatments of glioma include surgical resection, supplemented with radiotherapy and chemotherapy, but the prognosis is poor. PARP-1 (Poly ADP-ribose polymerase 1) is a hot spot for cancer-targeted therapy and was reported to be significantly elevated in glioma. In this study, we analyzed the role of PARP-1 in DNA damage repair, constructed a PARP1-related DNA-repair prognostic signature (DPS), and screened targeted drugs for glioma. RNA-seq data of 639 glioma samples were downloaded from the GEO (Gene Expression Omnibus) database and divided into PARP1_H and PARP1_L according to the front and rear thirds of the expression level of PARP-1. First, we systematically analyzed the influence of PARP-1 on DNA damage repair, prognosis, and chemoradiotherapy sensitization of glioma. All glioma patients and patients with radiotherapy or chemotherapy had a better prognosis in PARP1_L than in PARP1_H. Next, differentially expressed DNA-repair related genes (DEGs) were identified between PARP1_H and PARP1_L by LASSO (Least Absolute Shrinkage and Selection Operator) Cox analysis and applied for constructing DPS. Based on the four-gene DPS, we then developed a new nomogram to assess overall survival in glioma patients. Additionally, PARP-1 was proved an effective target for glioma therapy. So, a series of computer-aided techniques, including Discovery Studio 4.5, Schrodinger, and PyMol, were applied for the virtual screening of favorable PARP-1 inhibitors. In conclusion, this study investigated the effect of PARP-1 on glioma prognosis and the sensitization effect of radiotherapy and chemotherapy, established a novel nomogram to evaluate the overall survival of glioma patients, and further explored targeted therapy for glioma.

In this paper, we consider the problem of the $S$ -asymptotically $\omega $ -periodic synchronization of fractional-order complex-valued recurrent neural networks with time delays. Firstly, we can not explicitly decompose the fractional-order complex-valued systems into equivalent fractional-order real-valued systems, by means of the contraction mapping principle and some important features of Mittag-Leffler functions, we obtain some sufficient conditions for the existence and uniqueness of $S$ -asymptotically $\omega $ -periodic solutions for this class of neural networks. Then, by constructing an appropriate Lyapunov functional, the theory of fractional differential equation, and some inequality techniques, sufficient conditions are obtained to guarantee the global Mittag-Leffler synchronization of the drive-response systems. Finally, two examples are given to illustrate the effectiveness and feasibility of our main results.

Abstract Background Nanoparticle-based pulmonary drug delivery systems are commonly developed and applied for drug-targeted delivery. They exhibit significant advantages compared to traditional pulmonary drug delivery systems. However, developing the formulation of each drug is a time-consuming and laborious task. Results In this study, a universal lung-targeting nanoparticle was designed and constructed. The self-assembled micelles were composed of a platycodon secondary saponin, 3-O-β-d-glucopyranosyl platycodigenin 682 (GP-682), based on its specific amphiphilic structure. The GP-682 micelles exhibited a relatively stable zeta potential with a particle size between 60 and 90 nm, and the critical micelle concentration (CMC) value was approximately 42.3 μg/mL. Preincubation of GP-682 micelles markedly enhanced their cell membrane permeability and improved drug uptake in vitro. The results were visualized using fluorescent dye tracing, transmission electron microscopy (TEM) observations and the lactate dehydrogenase (LDH) release assay. The obtained benefits enhanced the distribution of levofloxacin (Lev) in mouse lung tissue and reduced antibiotics overdosing. The acute lung injury mouse model induced by the Pseudomonas aeruginosa PA 14 strain demonstrated that preinjection of GP-682 micelles before antibiotic administration resulted in a higher survival rate and anti-infective efficacy in vivo. It also caused reductions in pulmonary injury, bacterial invasion and cytokine expression compared with treatment with Lev alone. Conclusions GP-682 micelles are another nanoparticle-based pulmonary drug delivery system and provide a new lung-targeting therapy option.

Although the tumor suppressor P53 is known to regulate a broad network of signaling pathways, it is still unclear how certain drugs influence these P53 signaling networks. Here, we used a comprehensive single-cell multiomics view of the effects of ginsenosides on cancer cells. Transcriptome and proteome profiling revealed that the antitumor activity of ginsenosides is closely associated with P53 protein. A miRNA–proteome interaction network revealed that P53 controlled the transcription of at least 38 proteins, and proteome-metabolome profiling analysis revealed that P53 regulated proteins involved in nucleotide metabolism, amino acid metabolism and “Warburg effect”. The results of integrative multiomics analysis revealed P53 protein as a potential key target that influences the anti-tumor activity of ginsenosides. Furthermore, by applying affinity mass spectrometry (MS) screening and surface plasmon resonance fragment library screening, we confirmed that 20(S)-protopanaxatriol directly targeted adjacent regions of the P53 DNA-binding pocket and promoted the stability of P53–DNA interactions, which further induced a series of omics changes.

The automatic extraction of individual tree from mobile laser scanning (MLS) scenes has important applications in tree growth monitoring, tree parameter calculation and tree modeling. However, trees often grow in rows and tree crowns overlap with varying shapes, and there is also incompleteness caused by occlusion, which makes individual tree extraction a challenging problem. In this paper, we propose a trunk-constrained and tree structure analysis method to extract trees from scanned urban scenes. Firstly, multi-feature enhancement is performed via PointNet to segment the tree points from raw urban scene point clouds. Next, the candidate local tree trunk clusters are obtained by clustering based on the intercepted local tree trunk layer, and the real local tree trunk is obtained by removing noise data. Then, the trunk is located and extracted by combining circle fitting and region growing, so as to obtain the center of the tree crown. Further, the points near the tree’s crown (core points) are segmented through distance difference, and the tree crown boundary (boundary points) is distinguished by analyzing the density and centroid deflection angle. Therefore, the core and boundary points are deleted to obtain the remaining points (intermediate points). Finally, the core, intermediate and boundary points, as well as the tree trunks, are combined to extract individual tree. The performance of the proposed method was evaluated on the Pairs-Lille-3D dataset, which is a benchmark for point cloud classification, and data were produced using a mobile laser system (MLS) applied to two different cities in France (Paris and Lille). Overall, the precision, recall, and F1-score of instance segmentation were 90.00%, 98.22%, and 99.08%, respectively. The experimental results demonstrate that our method can effectively extract trees with multiple rows of occlusion and improve the accuracy of tree extraction.

Objective: By combining the expression profiles of metabolism-related genes (MRGS) with clinical information, the expression quantities of MRGS and the influence on development and prognosis were systematically analyzed, so as to provide a theoretical basis for the clinical study on the prognosis of Ewing's sarcoma. Methods: MRGs expression profiles of 64 patients with Ewing's sarcoma were obtained from GEO dataset. Univariate Cox regression analysis was used to identify metabolization-related differentially expressed genes (DEGs) related with prognosis in Ewing's sarcoma patients. Then, multivariate Cox analysis was used to calculate novel prognostic markers based on metabolism-related DEGs. Besides, We validate the model using ICGC datasets. Finally, the new prognostic index was verified on the basis of the prognostic models. Results: Multivariate Cox regression analysis identified 74 metabolization-related DEGs, 25 of which were associated with Ewing's sarcoma patients' overall survival. Subsequently, we used 25 DEGs to construct metabolism-related prognostic signature for patients with Ewing's sarcoma. Based on the 18 DEGs regression coefficient, we propose the formula of each patient's risk score, and then divided the patients into high-risk group and low-risk group. The results indicated that the survival rate and survival time were higher in the low-risk group and lower in the high-risk group. Multivariate Cox analysis showed that risk score index was an independent prognostic factor for Ewing's sarcoma. Conclusion: The experimental results suggest that the 18 metabolism-related DEGs marker may be effective in predicting the prognosis of Ewing's sarcoma to some extent, helping to individualize treatment of patients at different risks.

BackgroundHypothyroidism in the first trimester of pregnancy (T1) has great adverse effects on mothers and foetuses. However, few studies have investigated the influence on postpartum thyroid dysfunction. This study aimed to evaluate their long-term effect on postpartum thyroid function within one year after delivery.MethodsIn total, 151 women were recruited from 1496 participants and were classified as newly diagnosed subclinical hypothyroidism (SCH) in T1 (ND-SCH, n=50), previously known SCH before pregnancy (PK-SCH, n=51) and previously known overt hypothyroidism (PK-OH, n=50). Their thyroid functions were dynamically monitored from pre-conception to one-year postpartum.ResultsDuring pregnancy, the first thyroid functions’ test time in T1 were 5-8 gestational weeks. After delivery, the prevalence of postpartum thyroiditis (PPT) was comparable in women with previously known and newly diagnosed hypothyroidism [ND-SCH 62.0% vs PK-SCH 64.7% vs PK-OH 64.0%, P=0.96]. For the ND-SCH group, PPT was significantly related with thyroid-stimulating hormone (TSH) >4.0 mU/L occurring at 2.5 mU/L in T1 [OR=3.55, 95% CI, 1.43-8.81] and TPOAb≥300 μIU/mL [OR=6.58, 95% CI, 2.05-21.12] were associated with PPT. Regardless of whether SCH was diagnosed before pregnancy or in T1, the levothyroxine (LT4) treatment was discontinued at delivery. More than 50% of the patients had to face the hypothyroidism phase of postpartum and restarted LT4 treatment in the first-year follow-up. The logistic regression analysis revealed that TSH elevation occurring at

Puerarin, one of the major isoflavonoid compounds isolated from Gegen, is widely used to treat glucose and lipid disorders. Identifying the mechanism of puerarin’s action against lipid and glucose metabolism dysfunction is very important for its application and development. Based on the structure of puerarin, different chemical probes were synthesized to capture and identify puerarin’s protein target. A membrane translocation assay was conducted to identify the interaction between puerarin and target proteins. The results show that puerarin targets the PH domain of Akt1, inhibits Akt1′s transmembrane effect and activates Akt1. Activated Akt1 improves the phosphorylation or activity of downstream proteins, such as GSK-3β and PDE3, and ultimately regulates glucose and lipid metabolism. Puerarin, as a dietary supplement, could target the PH domain of Akt1 to activate the insulin pathway and improve insulin resistance, thereby assisting in the regulation of glucose and lipid metabolism disorders.

Members of the RAS proto-oncogene superfamily are indispensable molecular switches that play critical roles in cell proliferation, differentiation, and cell survival. Recent studies have attempted to prevent the interaction of RAS/GTP with RAS guanine nucleotide exchange factors (GEFs), impair RAS-effector interactions, and suppress RAS localization to prevent oncogenic signalling. The present study aimed to investigate the effect of the natural triterpenoic acid inhibitor glycyrrhetinic acid, which is isolated from the roots of Glycyrrhiza plant species, on RAS stability. We found that glycyrrhetinic acid may bind to the P-loop of RAS and alter its stability. Based on our biochemical tests and structural analysis results, glycyrrhetinic acid induced a conformational change in RAS. Meanwhile, glycyrrhetinic acid abolishes the function of RAS by interfering with the effector protein RAF kinase activation and RAS/MAPK signalling. KEY WORDS: Glycyrrhetinic acid, RAS, Allosteric inhibitor, RAS/MAPK signalling

Urban trees are vital elements of outdoor scenes via mobile laser scanning (MLS), accurate individual trees detection from disordered, discrete, and high-density MLS is an important basis for the subsequent analysis of city management and planning. However, trees cannot be easily extracted because of the occlusion with other objects in urban scenes. In this work, we propose a coarse-to-fine individual trees detection method from MLS point cloud data (PCD) based on treetop points extraction and radius expansion. Firstly, an improved semantic segmentation deep network based on PointNet is applied to segment tree points from the scanned urban scene, which combining spatial features and dimensional features. Next, through calculating the local maximum, the candidate treetop points are located. In addition, the optimized treetop points are extracted after the tree point projection plane was filtered to locate the candidate treetop points, and a distance rule is used to eliminate the pseudo treetop points then the optimized treetop points are obtained. Finally, after the initial clustering of treetop points and vertical layering of tree points, a top-down layer-by-layer segmentation based on radius expansion to realize the complete individual extraction of trees. The effectiveness of the proposed method is tested and evaluated on five street scenes in point clouds from Oakland outdoor MLS dataset. Furthermore, the proposed method is compared with two existing individual trees segmentation methods. Overall, the precision, recall, and F-score of instance segmentation are 98.33%, 98.33%, and 98.33%, respectively. The results indicate that our method can extract individual trees effectively and robustly in different complex environments.

Kaempferol and quercetin, as dietary flavonoids, are beneficial to diabetes risk reduction. Clarifying the mechanism of dietary flavonoids in improving glucose metabolism may meet the functional and nutritional requirements. In this paper, PCA revealed two glucuronides, kaempferol-3-glucuronide (K-3-G) and quercetin-3-glucuronide (Q-3-G), which were the primary metabolites. Target prediction, enzymatic activity, and AKT plasma translocation assays indicated that K-3-G and Q-3-G could target the AKT PH domain. Fluorescence spectrometry was used to measure the dissociation constants of K-3-G (KD = 27.6 μM) and Q-3-G (KD = 10.8 μM). Molecular docking provided interaction insights for K-3-G and Q-3-G with the AKT PH domain. K-3-G and Q-3-G activated the AKT/GSK3β phosphorylation, thereby improving glucose consumption. Our study demonstrates that the glucuronide metabolites of flavonoids directly target the AKT PH domain and activate AKT/GSK3β signal pathway, thereby improving glucose metabolism.

Background. Propofol is a widely used intravenous anesthetic drug with potential neuroprotective effect in diverse diseases of neuronal injuries such as traumatic brain injury and ischemic stroke. However, the underlying molecular mechanism remains largely unknown. Methods. Real-time qPCR, enzyme-linked immunosorbent assay, and Western blotting were used to identify the expression pattern of miR-221/222, inflammatory genes, cytokines, and IRF2. The biological roles and mechanisms of propofol in microglia activation were determined in BV2 cells and primary microglia. Bioinformatic analysis and luciferase reporter assay were used to confirm the regulatory role of miR-221/222 in Irf2 expression. Results. We found that miR-221 and miR-222 were downstream targets of propofol and were consistently upregulated in lipopolysaccharide- (LPS-) primed BV2 cells. Gain- and loss-of-function studies revealed that miR-221 and miR-222 were profoundly implicated in microglia activation. Then, interferon regulatory factor 2 (Irf2) was identified as a direct target gene of miR-221/222. IRF2 protein levels were reduced by miR-221/222 and increased by propofol treatment. Ectopic expression of IRF2 attenuated the proinflammatory roles induced by LPS in BV2 cells. More importantly, the suppressive effects of propofol on LPS-primed activation of BV2 cells or primary mouse microglia involved the inhibition of miR-221/222-IRF2 axis. Conclusions. Our study highlights the critical function of miR-221/222, which inhibited Irf2 translation, in the anti-inflammatory effects of propofol, and provides a new perspective for the molecular mechanism of propofol-mediated neuroprotective effect.

Background. Propofol is a known intravenous hypnotic drug used for induction and maintenance of sedation and general anesthesia. Emerging studies also reveal a neuroprotective effect of propofol in diverse diseases of neuronal injuries via modulating microglia activation. In this study, we aimed to uncover the downstream targets of propofol in this process. Methods. RNA sequencing analysis to identify genes implicated in the propofol-mediated neuroprotective effect. Quantitative real-time PCR, enzyme-linked immunosorbent assay, and Western blotting analysis were performed to analyze inflammatory gene expression, cytokine levels, and TGM2. BV2 cells and primary microglia were used for functional verification and mechanism studies. Results. The multifunctional enzyme transglutaminase 2 (TGM2) was identified as a putative functional mediator of propofol. TGM2 was significantly upregulated in lipopolysaccharide- (LPS-) primed BV2 cells. Genetic silencing of TGM2 abolished LPS-induced microglial activation. Notably, gain-of-function experiments showed that the proinflammatory effects of TGM2 were dependent on its GTP binding activity instead of transamidase activity. Then, TGM2 was revealed to activate the NF-κB signaling pathway to facilitate microglial activation. Propofol can inhibit TGM2 expression and NF-κB signaling in BV2 cells and primary microglia. Ectopic expression of TGM2 or constitutively active IKKβ (CA-IKKβ) can compromise propofol-induced anti-inflammatory effects. Conclusions. Our findings suggest that TGM2-mediated activation of NF-κB signaling is an important mechanism in the propofol-induced neuroprotective effect that prevents microglial activation.

Background: Previous studies indicate the effects of thyroid dysfunction on adverse obstetric outcomes and fetal neurodevelopment, of which the results on gestational anemia are controversial. Here, we evaluated the influence of thyroid dysfunction on gestational anemia via published epidemiological articles and a new prospective study conducted by our team, respectively.Methods: We searched studies on the PubMed, Embase, MEDLINE, and Cochrane databases as of November 2019, and conducted a prospective study in which participants underwent thyroid function and blood routine testing throughout pregnancy.Results: The meta-analysis showed that pregnancies with overt hypothyroidism [OH; odds ratio (OR) = 3.74, 95% confidence interval (CI): 1.95–7.15] or that were thyroid peroxidase antibody (TPOAb)-positive (OR = 1.97, 95%CI: 1.19–3.26) had increased anemia risk, but similar results were not found in pregnancies with subclinical hypothyroidism (SCH) and hyperthyroidism. In the prospective study from our new data, the hypothyroid group had significant reductions in hemoglobin (Hb) (P = 0.048) and increased anemia risk (OR = 6.384, 95%CI: 2.498–16.311) during the second half of pregnancy. From the first to second half of pregnancy, the longitudinal reductions in Hb, erythrocyte (RBC), and hematocrit (Hct) levels were significantly increased in hypothyroid group.Conclusions: Our meta-analysis indicates that untreated OH or TPOAb-positive pregnant women have increased risk of anemia. In addition, our new data showed that treated hypothyroidism is also a risk factor for anemia in the second half of pregnancy rather than in the first half. The results may guide strengthening of Hb monitoring in pregnancies with thyroid dysfunction.

Neural networks and deep learning have been successfully applied to tackle problems in drug discovery with increasing accuracy over time. There are still many challenges and opportunities to improve molecular property predictions with satisfactory accuracy even further. Here, we proposed a deep-learning architecture model, namely Bidirectional long short-term memory with Channel and Spatial Attention network (BCSA), of which the training process is fully data-driven and end to end. It is based on data augmentation and SMILES tokenization technology without relying on auxiliary knowledge, such as complex spatial structure. In addition, our model takes the advantages of the long- and short-term memory network (LSTM) in sequence processing. The embedded channel and spatial attention modules in turn specifically identify the prime factors in the SMILES sequence for predicting properties. The model was further improved by Bayesian optimization. In this work, we demonstrate that the trained BSCA model is capable of predicting aqueous solubility. Furthermore, our proposed method shows noticeable superiorities and competitiveness in predicting oil–water partition coefficient, when compared with state-of-the-art graphs models, including graph convoluted network (GCN), message-passing neural network (MPNN), and AttentiveFP.

Abstract Background Tracking targets of natural products is one of the most challenging issues in fields ranging from pharmacognosy to biomedicine. It is widely recognized that the biocompatible nanoparticle (NP) could function as a “key” that opens the target “lock”. Results We report a functionalized poly-lysine NP technique that can monitor the target protein of arctigenin (ATG) in vivo non-invasively. The NPs were synthesized, and their morphologies and surface chemical properties were characterized by transmission electron microscopy (TEM), laser particle size analysis and atomic force microscopy (AFM). In addition, we studied the localization of ATG at the level of the cell and the whole animal (zebrafish and mice). We demonstrated that fluorescent NPs could be ideal carriers in the development of a feasible method for target identification. The distributions of the target proteins were found to be consistent with the pharmacological action of ATG at the cellular and whole-organism levels. Conclusions The results indicated that functionalized poly-lysine NPs could be valuable in the multimodal imaging of arctigenin.

Platycodon grandiflorum, as a traditional medicinal plant, is commonly used in the treatment of pulmonary disease. Platycodon saponins are proposed as active ingredients. However, the role of secondary saponin metabolites (SSM) in the traditional use of Platycodon has not yet been fully clarified. In this study, [18F]-phillygenin ([18F]-PH) probe was synthesized and thereby used as a tracer for micro-positron emission tomography scanning to explore the effects of platycodon saponins. The membrane permeability with different SSM was evaluated in vitro based on the dye-carrying capacity of fluorescein isothiocyanate. The results showed that total platycodon saponins improved the dosimetry of [18F]-PH in the lung tissue, and an SSM named 3-O-β-D-glucopyranosyl platycodigenin (GPD682) appreciably changed the distribution of drugs both in vitro and in vivo. We propose that GPD682 could be utilized as an important ingredient to help drug delivery to the lung tissue and improve the treatment of respiratory disease.

Chronic obstructive pulmonary disease (COPD) has been a major public health problem and is still a formidable challenge for clinicians. It is urgent to find new compounds for minimizing the risk of disease progression and exacerbation especially in the early phase of COPD. A traditional Chinese medicine (TCM) formula, Chuan Bei Pi Pa dropping pills (CBPP), was tested in this study to investigate its potential mechanisms in preventing the exacerbation of COPD. Phosphoproteomics analysis for a smog stimulated early stage COPD mice model was employed to detect the underlying molecular mechanisms of CBPP. In addition, protein–protein interaction (PPI) and bioinformatics analyses were included to analyze the key proteins and predict the key bioactive compounds. The results indicated that peiminine (PEI) target epidermal growth factor receptor (EGFR) prevented the exacerbation of COPD by inhibiting the EGFR signaling pathway, and ursolic acid (UA) can alleviate inflammation disorders via inhibition of CASP3 on mitogen-activated protein kinase (MAPK) signaling pathway. After in vivo and in vitro evaluations, we revealed that PEI from CBPP, as a lead compound, can improve lung function and alleviate pulmonary fibrosis by acting on the EGFR and MLC2 signaling pathways. Furthermore, the approach described here is an effective way to analyze and identify the bioactive ingredients from a mixture by functional proteomics analysis.

The effects of calcium chloride (CaCl2) treatment on chilling injury (CI), reactive oxygen species (ROS) metabolism, and ascorbate-glutathione (AsA-GSH) cycle in loquat fruit at 1 °C storage for 35 d were investigated. The results indicated that CaCl2 treatment remarkably suppressed the increase in browning index and firmness as well as the decrease in extractable juice rate. CaCl2 treatment also decreased the production of superoxide radical (O2•−), hydrogen peroxide (H2O2) content, but increased the 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical (OH•) scavenging ability, the activities of superoxide dismutase (SOD), catalase (CAT), and their gene expressions. Moreover, compared to the control loquat fruit, CaCl2-treated fruit maintained higher contents of AsA, GSH, higher levels of activities of ascorbate peroxidase (APX), glutathione reductase (GR), dehydroascorbate reductase (DHAR), and monodehydroascorbate reductase (MDHAR) and expressions of EjAPX, EjGR, EjMDHAR, and EjDHAR, but exhibited lower glutathione disulfide (GSSG) content. These results suggested that CaCl2 treatment alleviated CI in loquat fruit through enhancing antioxidant enzymes activities and AsA-GSH cycle system to quench ROS.

Near-infrared spectroscopy (NIRS) with its fast and nondestructive advantages can be qualified for the real-time quantitative analysis. This paper demonstrates that NIRS combined with partial least squares (PLS) regression can be used as a rapid analytical method to simultaneously quantify l-glutamic acid (l-Glu) and γ-aminobutyric acid (GABA) in a biotransformation process and to guide the optimization of production conditions when the merits of NIRS are combined with response surface methodology. The high performance liquid chromatography (HPLC) reference analysis was performed by the o-phthaldialdehyde pre-column derivatization. NIRS measurements of two batches of 141 samples were firstly analyzed by PLS with several spectral pre-processing methods. Compared with those of the HPLC reference analysis, the resulting determination coefficients (R2), root mean square error of prediction (RMSEP) and residual predictive deviation (RPD) of the external validation for the l-Glu concentration were 99.5%, 1.62 g/L, and 11.3, respectively. For the GABA concentration, R2, RMSEP, and RPD were 99.8%, 4.00 g/L, and 16.4, respectively. This NIRS model was then used to optimize the biotransformation process through a Box-Behnken experimental design. Under the optimal conditions without pH adjustment, 200 g/L l-Glu could be catalyzed by 7148 U/L glutamate decarboxylase (GAD) to GABA, reaching 99% conversion at the fifth hour. NIRS analysis provided timely information on the conversion from l-Glu to GABA. The results suggest that the NIRS model can not only be used for the routine profiling of enzymatic conversion, providing a simple and effective method of monitoring the biotransformation process of GABA, but also be considered to be an optimal tool to guide the optimization of production conditions.

Gram-negative pathogen–induced nosocomial infections and resistance are a most serious menace to global public health. Qingfei Xiaoyan Wan (QF), a traditional Chinese medicine (TCM) formula, has been used clinically in China for the treatment of upper respiratory tract infections, acute or chronic bronchitis and pulmonary infection. In this study, the effects of QF on Pseudomonas aeruginosa–induced acute pneumonia in mice were evaluated. The mechanisms by which four typical anti-inflammatory ingredients from QF, arctigenin (ATG), cholic acid (CLA), chlorogenic acid (CGA) and sinapic acid (SPA), regulate anti-inflammatory signaling pathways and related targets were investigated using molecular biology and molecular docking techniques. The results showed that pretreatment with QF significantly inhibits the release of cytokines (TNF-α and IL-6) and chemokines (IL-8 and RANTES), reduces leukocytes recruitment into inflamed tissues and ameliorates pulmonary edema and necrosis. In addition, ATG was identified as the primary anti-inflammatory agent with action on the PI3K/AKT and Ras/MAPK pathways. CLA and CGA enhanced the actions of ATG and exhibited synergistic NF-κB inactivation effects possibly via the Ras/MAPK signaling pathway. Moreover, CLA is speculated to target FGFR and MEK firstly. Overall, QF regulated the PI3K/AKT and Ras/MAPK pathways to inhibit pathogenic bacterial infections effectively.

Central composite design (CCD), utilized with three independent variables, verified that the optimal treatment conditions in bitter melon fruit were melatonin (MT) concentration of 120 µmol L−1, ethanol concentration of 6%, and immersing time of 10 min. Under optimal conditions, the experimental values of firmness, chilling injury (CI) index, and weight loss were shown as 27.81 N, 65.625%, and 0.815%, respectively. Moreover, the combined effect of MT and ethanol on CI and physiological quality in postharvest bitter melon fruit stored at 4 °C was investigated. It was found that the combined treatment contributed to the reduced CI symptoms and inhibited ion leakage and malondialdehyde (MDA) accumulation. Moreover, higher levels of chlorophyll, total soluble solids (TSSs), soluble sugar, soluble protein, and ascorbic acid (AsA) were observed in comparison with the control group. Furthermore, the synthesis of total phenols and flavonoids in bitter melon was greatly promoted. Therefore, the combination of MT and ethanol could have the potential for alleviating CI and maintaining postharvest quality for the duration of cold storage.

BackgroundInflammasomes, which mediate maturation of interleukin-1β (IL-β) and interleukin-18 (IL-18) and lead to pyroptosis, have been linked to various autoimmune disorders. This study investigated whether they are involved in the pathogenesis of autoimmune thyroiditis (AIT).MethodsWe collected thyroid tissues from 50 patients with AIT and 50 sex- and age-matched controls. Serum levels of free T3, free T4, thyrotropin, thyroid peroxidase antibody (TPOAb), and thyroglobulin antibody (TgAb) were measured by electrochemiluminescent immunoassays. Expression of several inflammasome components, the NOD-like receptor (NLR) family pyrin domain containing 1 (NLRP1), NLRP3, CARD-domain containing 4 (NLRC4), absent in melanoma 2 (AIM2), the apoptosis-associated speck-like protein that contains a caspase recruitment domain (ASC), caspase-1, IL-1β, and IL-18 was determined by real-time PCR and western blot. Immunohistochemistry was used to localize the expression of NLRP1, NLRP3, NLRC4, and AIM2. The Nthy-ori 3-1 thyroid cell line was stimulated with tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-17A, interleukin-6, and poly(dA:dT). The levels of IL-18 and IL-1β in the cell supernatant were measured by enzyme-linked immunosorbent assay, and lactate dehydrogenase was quantified by absorptiometry. ASC specks were examined by confocal immunofluorescence microscopic analysis. Cell death was examined by flow cytometry, and the N-terminal domain of gasdermin D was detected by western blot analysis.ResultsExpression of NLRP1, NLRP3, NLRC4, AIM2, ASC, caspase-1, pro IL-1β, pro IL-18, mRNA, and protein was significantly increased in thyroid tissues from patients with AIT, and enhanced posttranslational maturation of caspase-1, IL-18 and IL-1β was also observed. Expression of NLRP1, NLRP3, NLRC4, and AIM2 was localized mainly in thyroid follicular cells adjacent to areas of lymphatic infiltration. The thyroid mRNA level of NLRP1 and ASC was correlated to the serum TPOAb and TgAb levels in the AIT group. TNF-α and IFN-γ had a priming effect on the expression of multiple inflammasome components in thyroid cells. IFN-γ was found to strengthen poly(dA:dT)-induced cell pyroptosis and bioactive IL-18 release.ConclusionOur work has demonstrated for the first time that multiple inflammasomes are associated with AIT pathogenesis. The identified NLRP3, NLRP1, NLRC4, AIM2 inflammasomes and their downstream cytokines may represent potential therapeutic targets and biomarkers of AIT.

Classical Chinese pharmacopeias describe numerous excellent herbal formulations, and each prescription is an outstanding pool of effective compounds for drug discovery. Clarifying the bioactivity of the combined mechanisms of the ingredients in complex traditional Chinese medicine formulas is challenging. A classical formula known as Qingfei Xiaoyan Wan, used clinically as a treatment for prevalent chronic lung disease, was investigated in this work. A mutually enhanced bioactivity-guided ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS) characterization system was proposed, coupled with a dual-luciferase reporter assay for β2AR-agonist cofactor screening. Arctiin, arctigenin, descurainoside and descurainolide B, four lignin compounds that showed synergistic bronchodilation effects with ephedrine, were revealed. The synergistic mechanism of arctigenin with the β2ARagonist involved with the reduction of free Ca2+ was clarified by a dual-luciferase reporter assay for intracellular calcium and the Ca2+ indicator fluo-4/AM to monitor changes in the fluorescence. The relaxant and contractile responses of airway smooth muscle are regulated by crosstalk between the intracellular cAMP and calcium signaling pathways. Our data indicated the non-selective βAR agonist ephedrine as the principal bronchodilator of the formula, whereas the lignin ingredients served as adjuvant ingredients. A greater understanding of the mechanisms governing the control of these pathways, based on conventional wisdom, could lead to the identification of novel therapeutic targets or new agents for the treatment of asthma and COPD.

Lotus nelumbo (LN) (Nelumbo nucifera Gaertn.) is an aquatic crop that is widely distributed throughout Asia and India, and various parts of this plant are edible and medicinal. It is noteworthy that different organs of this plant are used in traditional herbal medicine or folk recipes to cure different diseases and to relieve their corresponding symptoms. The compounds that are contained in each organ, which are named based on their chemical compositions, have led to their respective usages. In this work, a strategy was used to identify the difference ingredients and screen for Nuclear-factor-kappaB (NF-κB) inhibitors with anti-inflammatory ability in LN. Seventeen main difference ingredients were compared and identified from 64 samples of 4 different organs by ultra-performance liquid chromatography that was coupled with quadrupole/time of flight mass spectrometry (UPLC/Q-TOF-MS) with principal component analysis (PCA). A luciferase reporter assay system combined with the UPLC/Q-TOF-MS information was applied to screen biologically active substances. Ten NF-κB inhibitors from Lotus plumule (LP) extracts, most of which were isoquinoline alkaloids or flavone C-glycosides, were screened. Heat map results showed that eight of these compounds were abundant in the LP. In conclusion, the LP extracts were considered to have the best anti-inflammatory ability of the four LN organs, and the chemical material basis (CMB) of this biological activity was successfully validated by multivariate statistical analysis and biological research methods.

Glycyrrhetic acid (GA) exerts synergistic anti-asthmatic effects via a β₂-adrenergic receptor (β₂AR)-mediated pathway. Cholesterol is an important component of the structure and function of lipid rafts, which play critical roles in the β₂AR-Gs-adenylate cyclase (AC)-mediated signaling pathway. Owing to the structural similarities between GA and cholesterol, we investigated the possibility that GA enhances β₂AR signaling by altering cholesterol distribution. Azide-terminal GA (ATGA) was synthesized and applied to human embryonic kidney 293 (HEK293) cells expressing fusion β₂AR, and the electron spin resonance (ESR) technique was utilized. GA was determined to be localized predominantly on membrane and decreased their cholesterol contents. Thus, the fluidity of the hydrophobic region increased but not the polar surface of the cell membrane. The conformations of membrane proteins were also changed. GA further changed the localization of Gαs from lipid rafts to non-raft regions, resulting the binding of β₂AR and Gαs, as well as in reduced β₂AR internalization. Co-localization of β₂AR, Gαs, and AC increased isoproterenol-induced cAMP production and cholesterol reloading attenuated this effect. A speculation wherein GA enhances beta-adrenergic activity by increasing the functional linkage between the subcomponents of the membrane β₂AR-protein kinase A (PKA) signaling pathway was proposed. The enhanced efficacy of β₂AR agonists by this novel mechanism could prevent tachyphylaxis.