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Background. 4-Methoxydalbergione (4MOD) is a flavonoid isolated from the heartwood of Dalbergia. Studies have demonstrated that 4MOD exerts anticancer activities on bladder cancer and astrocytoma. However, the anticancer activity of 4MOD in hepatocellular carcinoma (HCC) remains unknown. This study aims to examine its anticancer activities and mechanisms in human liver cancer cells. Methods. CCK-8, colony forming, wound healing, transwell migration, and AnnexinV/PI assays were used to assess the anticancer effects of 4MOD in HCC cells. RNA sequencing (RNA-Seq) was selected to explore the possible mechanisms underlying the anti-HCC activity of 4MOD. The mRNA expression levels of target genes were verified through quantitative real-time PCR (qRT-PCR). A lentiviral shRNA interference technique was used to silence GADD45G expression. GADD45G knockdown was employed to confirm the crucial role of GADD45G in the 4MOD-mediatedanti-HCC effects. Results. 4MOD inhibited HCC cells’ proliferation and migration and promoted tumor cell apoptosis. RNA-Seq and qRT-PCR analyses revealed that 4MOD treatment increased GADD45G expression. Silencing GADD45G reversed 4MOD-mediated inhibition of proliferation, migration, and promotion of apoptosis. Conclusions. Our findings show that 4MOD elicits anti-HCC effects by upregulating GADD45G expression and could be a valuable anticancer agent for liver cancer.

In obesity, CD11c+ innate immune cells are recruited to adipose tissue and create an inflammatory state that causes both insulin and catecholamine resistance. We found that ablation of Gnas, the gene that encodes Gas, in CD11c expressing cells protects mice from obesity, glucose intolerance, and insulin resistance. Transplantation studies showed that the lean phenotype was conferred by bone marrow-derived cells and did not require adaptive immunity. Loss of cAMP signaling was associated with increased adipose tissue norepinephrine and cAMP signaling, and prevention of catecholamine resistance. The adipose tissue had reduced expression of catecholamine transport and degradation enzymes suggesting that the elevated norepinephrine resulted from decreased catabolism. Collectively, our results identified an important role for cAMP signaling in CD11c+ innate immune cells in whole-body metabolism by controlling norepinephrine levels in WAT, modulating catecholamine-induced lipolysis, and increasing thermogenesis, which together created a lean phenotype.

In obesity, CD11c+ innate immune cells are recruited to adipose tissue and create an inflammatory state that causes both insulin and catecholamine resistance. We found that ablation of Gnas, the gene that encodes Gαs, in CD11c expressing cells protects mice from obesity, glucose intolerance, and insulin resistance. Transplantation studies showed that the lean phenotype was conferred by bone marrow-derived cells and did not require adaptive immunity. Loss of cAMP signaling was associated with increased adipose tissue norepinephrine and cAMP signaling, and prevention of catecholamine resistance. The adipose tissue had reduced expression of catecholamine transport and degradation enzymes suggesting that the elevated norepinephrine resulted from decreased catabolism. Collectively, our results identified an important role for cAMP signaling in CD11c+ innate immune cells in whole-body metabolism by controlling norepinephrine levels in WAT, modulating catecholamine-induced lipolysis, and increasing thermogenesis, which together created a lean phenotype.

Supplemental Methods

Figure S1-7, Table S1

Hypoxia is known to be commonly present in breast tumor microenvironments. Stem-like cells that repopulate breast tumors, termed tumor-repopulating cells (TRC), thrive under hypoxic conditions, but the underlying mechanism remains unclear. Here, we show that hypoxia promotes the growth of breast TRCs through metabolic reprogramming. Hypoxia mobilized transcription factors HIF1α and FoxO1 and induced epigenetic reprogramming to upregulate cytosolic phosphoenolpyruvate carboxykinase (PCK1), a key enzyme that initiates gluconeogenesis. PCK1 subsequently triggered retrograde carbon flow from gluconeogenesis to glycogenesis, glycogenolysis, and the pentose phosphate pathway. The resultant NADPH facilitated reduced glutathione production, leading to a moderate increase of reactive oxygen species that stimulated hypoxic breast TRC growth. Notably, this metabolic mechanism was absent in differentiated breast tumor cells. Targeting PCK1 synergized with paclitaxel to reduce the growth of triple-negative breast cancer (TNBC). These findings uncover an altered glycogen metabolic program in breast cancer, providing potential metabolic strategies to target hypoxic breast TRCs and TNBC.Significance:Hypoxic breast cancer cells trigger self-growth through PCK1-mediated glycogen metabolism reprogramming that leads to NADPH production to maintain a moderate ROS level.

Background Individuals with certain chronic inflammatory lung diseases have a higher risk of developing lung cancer (LC). However, the underlying mechanisms remain largely unknown. Here, we hypothesized that chronic exposure to house dust mites (HDM), a common indoor aeroallergen associated with the development of asthma, accelerates LC development through the induction of chronic lung inflammation (CLI). Methods The effects of HDM and heat-inactivated HDM (HI-HDM) extracts were evaluated in two preclinical mouse models of LC (a chemically-induced model using the carcinogen urethane and a genetically-driven model with oncogenic KrasG12D activation in lung epithelial cells) and on murine macrophages in vitro. Pharmacological blockade or genetic deletion of the Nod-like receptor family pyrin domain-containing protein 3 (NLRP3) inflammasome, caspase-1, interleukin-1β (IL-1β), and C–C motif chemokine ligand 2 (CCL2) or treatment with an inhaled corticosteroid (ICS) was used to uncover the pro-tumorigenic effect of HDM. Results Chronic intranasal (i.n) instillation of HDM accelerated LC development in the two mouse models. Mechanistically, HDM caused a particular subtype of CLI, in which the NLRP3/IL-1β signaling pathway is chronically activated in macrophages, and made the lung microenvironment conducive to tumor development. The tumor-promoting effect of HDM was significantly decreased by heat treatment of the HDM extract and was inhibited by NLRP3, IL-1β, and CCL2 neutralization, or ICS treatment. Conclusions Collectively, these data indicate that long-term exposure to HDM can accelerate lung tumorigenesis in susceptible hosts (e.g., mice and potentially humans exposed to lung carcinogens or genetically predisposed to develop LC).

This study attempts to improve the vibration isolation performance of a vehicle suspension system with a magnetorheological damper (MRD) under complex driving conditions. Structure parameter uncertainty, disturbance of the driving process, and response time delay of MRD are all addressed. Firstly, experiments of MRD were carried out in a damping force testing machine to identify the parameters of the MRD adjustable Sigmoid model by the Levenberg-Marquardt optimization algorithm. Then, the parameter identification is verified by comparing experimental and simulation data. Secondly, the state space equations of the suspension system are derived by Newton’s second law. The transfer function from the bounded disturbance input to the control output is obtained based on H∞ control theory. To make the Infinite norm of the system transfer function less than a certain value, three control strategies are proposed: variable structure control (VSC), disturbance rejection control (DRC), and delay tolerance control (DTC). Thirdly, considering these issues together to weaken the effect of disturbances on vehicle driving conditions, a fuzzy cooperative control (FCC) strategy is proposed based on the linear matrix inequality (LMI) theory. Simulation results demonstrate that FCC semi-active vehicle suspension systems conduct effective vibration isolation performance while responding to multiple external disturbances.

The increasing availability of both indoor positioning services and sensors for 3D data capture, such as RGB-D sensors, allows the provision of indoor spatial information services for indoor localization-based applications. To efficiently realize these services, the indoor information and the relationships between indoor spaces are required. The recently released Indoor Geography Markup Language (IndoorGML) attempts to represent and exchange geo-information for modeling topology and semantics of indoor spaces. However, it is still challenging to map indoor space features to the IndoorGML-encoded navigation network model directly from colorized 3D points. Therefore, we propose a semantics-guided method for indoor navigation element reconstruction from RGB-D sensor data. First, a hierarchical indoor scene interpretation framework is used for robustly recognizing the architecture structures and doors, respectively. In the developed hierarchical structure, a graph convolutional network-based architectural structure recognition method is adopted to deduce the long-range interactions among primitives for describing the rich physical relationships in the real world. Its output is the produced initial annotated results, from which doors as the common openings are further detected using a U-Net-based door recognition method. This enables to effectively provide the semantic guidance for the cellular representation of the indoor space and its topological reconstruction. Second, an adaptive architectural structure-guided room segmentation method is developed by combining distance transform and watershed segmentation to determine cellular spaces according to the definition in IndoorGML. Third, taking the different states of doors into consideration, a door-guided topological relationship reconstruction method is proposed to achieve the network graph representation of indoor environments. In this context, a simulated door model is designed to correct and update the true position of a door leaf, and a virtual door is defined to optimize the topological analysis. As a consequence, an IndoorGML-encoded navigation network model is generated, which can be used as the base for indoor navigation applications independent of the platform. Experiments are performed on the public Stanford large-scale 3D Indoor Spaces Dataset to verify the robustness and effectiveness of the proposed method both qualitatively and quantitatively. Results indicate the capability of the proposed method in automatically reconstructing indoor navigation elements of Manhattan-world indoor environments from RGB-D sensor data.

Infectious microbes that spread easily in healthcare facilities remain as the severe threat for the public health, especially among immunocompromised populations. Given the intricate problem of dramatic increase in resistance to common biocides, the development of safe and efficient biocide formulated agents to alleviate drug resistance is highly demanding. In this study, Schiff-base ligands were successfully formed on natural biopolymer of epsilon-poly-L-lysine (e-PL) decorated aldehyde functionalized mesoporous silica SBA-15 (CHO-SBA-15) for the selective coordination of silver ions, which was affirmed by various physicochemical methods. Besides the identified broad-spectrum antibacterial activities, the as-prepared Schiff-base silver nanocomplex (CHO-SBA-15/e-PL/Ag, CLA-1) exhibited an improved inhibitory effect on infectious pathogen growth typified by Escherichia coli and Staphylococcus aureus in comparison with two control silver complexes without Schiff-base conjugates, SBA-15/e-PL/Ag and CHO-SBA-15/Ag, respectively. In addition, CLA-1 remarkably inhibited the growth of Mycobacterium tuberculosis due to the excellent antimicrobial activity of silver species. Significantly, CLA-1 kills Candida albicans cells, inhibits biofilm formation, and eliminates preformed biofilms, with no development of resistance during continuous serial passaging. The antifungal activity is connected to disruption of bacterial cell membranes and increased levels of intracellular reactive oxygen species. In mouse models of multidrug-resistant C. albicans infection, CLA-1 exhibited efficient in vivo fungicidal efficacy superior to two antifungal drugs, amphotericin B and fluconazole. Moreover, CLA-1 treatment induces negligible toxicity against normal tissues with safety. Therefore, this study reveals the pivotal role of the molecular design of Schiff-base silver nanocomplex formation on biopolymer surface-functionalized silica mesopores as a green and efficient nanoplatform to tackle infectious microbes.

BACKGROUND: This study aims to identify the feasibility of uniportal video-assisted thoracoscopic surgery (VATS) and robot-assisted thoracoscopic surgery (RATS) compared with multiportal VATS in the resection of mediastinal lesions. METHODS: Patients who underwent mediastinal lesion resection were enrolled and allocated to the uni-VATS, tri-VATS, and RATS groups according to the surgical approach. Propensity score-matched (PSM) analysis was performed between the VATS and RATS groups as well as the uni-VATS and tri-VATS groups. The operative and recovery parameters were compared. RESULTS: Totally, 274 patients were enrolled. There was no difference in the operative parameters among the groups. Compared with multiportal VATS, uniportal VATS and RATS had a significantly shorter chest tube placement time (2.43±0.88 vs. 1.78±1.22 vs. 2.21±1.11 days, P

An unprecedented dimeric pimarane-derived diterpenoid (trichanthol A, 1) and a new 17-nor-(9β-H)-pimarane diterpenoid glucoside (trichanthol B, 2) together with ten known analogs (3–12) were obtained from the tuber of Icacina trichantha (Icacinaceae) in a bioassay-guided phytochemical investigation. Trichanthol A (1) is the first example of a pimarane-derived diterpenoid dimer furnished by forming an undescribed C-16–C-7′ linkage, and this is the first report of the glucosylation of a 17-nor-(9β-H)-pimarane diterpenoid (2). Their structures were elucidated by the interpretation of spectroscopic data and chemical methods in combination with calculated 13C NMR-DP4+ analysis and electronic circular dichroism methods. All isolated compounds exhibited antibacterial activity against standard and drug-resistant Helicobacter pylori strains with MIC values ranging from 8 to 64 μg mL−1. Moreover, icacinlactone B (6) showed an additive effect in combination with metronidazole or clarithromycin against H. pylori.

Background Although the outcome of breast cancer patients has been improved by advances in early detection, diagnosis and treatment. Due to the heterogeneity of the disease, prognostic assessment still faces challenges. The accumulated data indicate that there is a clear correlation between the tumor immune microenvironment and clinical outcomes. Objective Construct immune-related gene pairs to evaluate the prognosis of breast cancer and patient survival rate. Methods From the Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) database, the Gene expression profiles and clinical data of breast cancer samples were downloaded. TCGA cohort were further divided into a training set (n = 764) and internal validation sets (n = 325). The GEO cohort was analyzed as an external validation cohort (n = 327). In the training set, differently expressed immune-relevant genes (IRGs) were screened firstly, and they were used to construct immune-relevant gene pairs (IRGPs). Then, the prognostic IRGPs were identified via univariate Cox regression analysis. Finally, least absolute shrinkage and selection operator (LASSO) Cox regression analysis was used to constituted the IRGP prognostic signature. Kaplan-Meier (KM) survival curves, receiver operating characteristic (ROC) curve analysis, univariate and multivariate Cox regression analysis were used to estimate the predictive value of the IRGP prognostic signature. And the IRGP prognostic signature was validated in the internal validation cohort and external validation cohort. We used gene set enrichment analysis (GSEA) to elucidate the biological functions of the IRGP prognostic signature. Results A total of 474 differently expressed IRGs and 2942 prognostic IRGPs were identified. Finally, we generated a IRGP prognostic signature consisting of 33 IRGPs. Subsequently, the 33 IRGPs grouped BRCA patients into high- and low-risk groups. Kaplan-Meier curves shown a significantly different overall survival in risk groups. Time-dependent ROC curves indicated that the IRGP prognostic signature possessed a high specificity and sensitivity in all the sets. Univariate and multivariate Cox regression analysis showed a statistical significance for the prognostic value of IRGP prognostic signature and the IRGP prognostic signature was a strong independent risk factor. The functional enrichment analysis indicated that low IRGP value was correlated with biological processes related to immune. Immune cell infiltration analysis indicated a significant difference in percentage of M2 macrophages between high- and low-risk groups. Conclusion The 33-IRGPs prognostic signature was developed to provide new insights for the identification of high-risk breast cancer and the evaluation of the possibility of immunotherapy in personalized breast cancer treatment.

Aspergillus fumigatus is a well-known opportunistic pathogen that causes invasive aspergillosis (IA) infections with high mortality in immunosuppressed individuals. Morphogenesis, including hyphal growth, conidiation, and cell wall biosynthesis is crucial in A. fumigatus pathogenesis. Based on a previous random insertional mutagenesis library, we identified the putative polysaccharide synthase gene Afcps1 and its para-log Afcps2. Homologs of the cps gene are commonly found in the genomes of most fungal and some bacterial pathogens. Afcps1/cpsA is important in sporulation, cell wall composition, and virulence. However, the precise regulation patterns of cell wall integrity by Afcps1/cpsA and further effects on the immune response are poorly understood. Specifically, our in-depth study revealed that Afcps1 affects cell-wall stability, showing an increased resistance of ΔAfcps1 to the chitinmicrofibril destabilizing compound calcofluor white (CFW) and susceptibility of ΔAfcps1 to the β-(1,3)-glucan synthase inhibitor echinocandin caspofungin (CS). Additionally, deletion of Afcps2 had a normal sporulation phenotype but caused hypersensitivity to Na+ stress, CFW, and Congo red (CR). Specifically, quantitative analysis of cell wall composition using high-performance anion exchange chromatography-pulsed amperometric detector (HPAEC-PAD) analysis revealed that depletion of Afcps1 reduced cell wall glucan and chitin contents, which was consistent with the down-regulation of expression of the corresponding biosynthesis genes. Moreover, an elevated immune response stimulated by conidia of the ΔAfcps1 mutant in marrow-derived macrophages (BMMs) during phagocytosis was observed. Thus, our study provided new insights into the function of polysaccharide synthase Cps1, which is necessary for the maintenance of cell wall stability and the adaptation of conidia to the immune response of macrophages in A. fumigatus.

External sensors are vulnerable to the external magnetic field, temperature, noise, and other factors in the semi-active control system based on magnetorheological (MR) damper, which not only reduces the reliability and stability but also increases the complexity, installation space, and system cost. Meanwhile, mechanical energy is generally dissipated as heat due to the friction between the cylinder and damper piston of the MR damper. To further broaden the applications of MR damper, it is essential to maintain excellent functions when the power supply is cut off in various emergencies. Based on this, a novel MR damper with displacement self-sensing and energy harvesting capability is proposed to solve the problems such as large structural size, high system cost, and vibration energy dissipation. Firstly, the structures of the MR damping, displacement self-sensing, and vibration energy harvesting are designed and integrated into the proposed MR damper. Then, the distribution of the magnetic flux density, displacement self-sensing voltage, and energy harvesting efficiency with a single induction coil and double induction coil is obtained using the finite element method. The experimental test system is set up and the performance of MR damping, displacement self-sensing voltage, and energy harvesting efficiency is experimentally tested. The experimental results show that the damping force reaches 513 N at the applied current of 0.25 A. At the same time, the amplitude of self-sensing voltage increases linearly with the increase of the amplitude of sinusoidal displacement excitation, and the displacement sensitivity comes up to 54.37 mV/mm. Moreover, the self-sensing voltage of the vibration energy harvesting component with a double induction coil is 2.512 V, and the energy harvesting efficiency is about twice than that with a single induction coil. It is found that the proposed MR damper, with a relatively compact structure and lower cost compared with traditional MR damper, not only promisingly achieves excellent dynamic damping performance but also simultaneously possesses displacement self-sensing ability and energy harvesting capability.

Dysbiosis of gut microbiota has been implicated in the pathogenesis of hypertension. A definite relationship between gut microbiota and hypertension remains intriguing. Here, we show that the Sacha inchi (Plukenetia volubilis L.) shell extract (SISE) intervention significantly reduced systolic blood pressures in spontaneous hypertensive rats (SHR), attenuated the oxidative damage and modulated plasma calcium homeostasis and left ventricular hypertrophy in both SHR and high-salt diet Wistar-Kyoto rats. SISE reshaped the gut microbiome and metabolome, particularly by improving the prevalence of Roseburia and dihydrofolic acid levels in the gut. Transcriptome analyses showed that the protective effects of SISE were accompanied by the modulation of renal molecular pathways, beneficial for cardiovascular functions such as the L-type voltage-dependent calcium channel (LTCC), a key regulator of calcium signaling. Overall, the results have shown that dietary SISE can alleviate hypertension regulating the gut microbiota, and Ca2+ signaling might be a potential target for spontaneous hypertension.

CD11c+ immune cells are a potential therapeutic target for treatment of obesity-related insulin resistance and type 2 diabetes (T2D). In obesity, CD11c+ immune cells are recruited to white adipose tissue and create an inflammatory state that causes both insulin and catecholamine resistance. In this study, we found that ablation of Gnas, the gene that encodes Gas, in CD11c expressing cells protects mice from high-fat diet-induced obesity, glucose intolerance and insulin resistance. GnasΔCD11c mice (KO) had increased oxygen consumption, energy expenditure, and beigeing of white adipose tissue (WAT). Transplantation studies showed that the lean phenotype was conferred by bone marrow-derived cells and the absence of T and B cells by crossing the KO to a Rag1-/- background did not alter the phenotype. Notably, we observed elevated norepinephrine and elevated cAMP signaling in the WAT of KO mice. The KO adipose tissue also had reduced expression of catecholamine transport and degradation enzymes. Collectively, our results identified an important role of Gas in CD11c+ cells in whole body metabolism regulation by controlling norepinephrine levels in WAT, modulating catecholamine-induced lipolysis and increasing thermogenesis that together created a lean phenotype.

Zhang et al. (2022) show that TCR signaling promotes the phosphorylation and activation of glycogen phosphorylase B (PYGB) in CD8(+) memory T (Tmem) cells. PYGB-dependent glycogen mobilization provides a carbon source to support glycolysis and early Tmem cell recall responses.

The prevalence of invasive aspergillosis with azole resistance is increasing, but the mechanisms underlying the development of resistance and treatment strategies are still limited. The present work is focused on finding a relationship between long-chain unsaturated fatty acids (LCUFAs), Aspergillus fumigatus development, and antifungal resistance. The effects of LCUFAs on antifungal agents in vitro were determined, and the stearic acid desaturase gene (sdeA) of A. fumigatus was characterized. In in vitro antifungal tests, LCUFAs antagonized the antifungal activity of itraconazole by extracting it from media, thereby preventing it from entering cells. The OA auxotrophic phenotype caused by an sdeA deletion confirmed that SdeA was required for OA biosynthesis in A. fumigatus. Furthermore, several low-level sdeA-overexpressing mutants with impaired vegetative growth phenotypes were successfully constructed. Additionally, an sdeA-overexpressing mutant, OEsdeA-5, showed lowered sensitivity levels to itraconazole. Moreover, RNA sequencing of OEsdeA-5 revealed that the altered gene-expression pattern. Through targeted metabolomics, decreased palmitic acid and stearic acid contents, accompanied by higher palmitoleic acid, margaroleic acid, and OA production levels, were found in OEsdeA-5. This study provides a novel insight of understanding of azole resistance and a potential target for drug development.

SummaryPlants employ an array of intricate and hierarchical signaling cascades to perceive and transduce informational cues to synchronize and tailor adaptive responses. Systemic stress response (SSR) is a recognized complex signaling and response network quintessential to plant’s local and distal responses to environmental triggers, however, the identity of the initiating signals has remained fragmented.Here, we show that both biotic (aphids and viral pathogens) and abiotic (high-light and wounding) stresses induce accumulation of the plastidial-retrograde-signaling metabolite, methylerythritol cyclodiphosphate (MEcPP), leading to reduction of the phytohormone, auxin, and the subsequent decreased expression of the phosphatase, PP2C.D1.This enables phosphorylation of mitogen-activated protein kinases (MAPK3/6), and the consequential induction of the downstream events ultimately resulting in biosynthesis of the two SSR priming metabolites, pipecolic- and N-hydroxy-pipecolic acid.This work identifies plastids as the initiation site, and the plastidial retrograde-signal, MEcPP as the initiator of a multi-component signaling cascade potentiating the biosynthesis of SSR activators, in response to biotic and abiotic triggers.

Glycogen has long been considered to have a function in energy metabolism. However, our recent study indicated that glycogen metabolism, directed by cytosolic phosphoenolpyruvate carboxykinase Pck1, controls the formation and maintenance of CD8+ memory T (Tmem) cells by regulating redox homeostasis1. This unusual metabolic program raises the question of how Pck1 is upregulated in CD8+ Tmem cells. Here, we show that mitochondrial acetyl coenzyme A is diverted to the ketogenesis pathway, which indirectly regulates Pck1 expression. Mechanistically, ketogenesis-derived β-hydroxybutyrate is present in CD8+ Tmem cells; β-hydroxybutyrate epigenetically modifies Lys 9 of histone H3 (H3K9) of Foxo1 and Ppargc1a (which encodes PGC-1α) with β-hydroxybutyrylation, upregulating the expression of these genes. As a result, FoxO1 and PGC-1α cooperatively upregulate Pck1 expression, therefore directing the carbon flow along the gluconeogenic pathway to glycogen and the pentose phosphate pathway. These results reveal that ketogenesis acts as an unusual metabolic pathway in CD8+ Tmem cells, linking epigenetic modification required for memory development. Zhang et al. show that ketogenesis-derived β-hydroxybutyrate (BHB) epigenetically modifies H3K9 of Foxo1 and Ppargc1a to regulate Pck1, which in turn controls metabolic flux and CD8+ memory T-cell development.

Background The development of minimally invasive surgical approaches has revolutionized surgical care and greatly improved surgical outcomes. This study aimed to evaluate the clinical effectiveness of a powered vascular stapler (PVS) during video-assisted thoracoscopic surgery (VATS) lobectomy. Methods This prospective, multi-center, post-market clinical study in China enrolled 50 patients with either a suspected or formal diagnosis of clinical stage IA to IIB non-small cell lung cancer (NSCLC) scheduled for VATS lobectomy. The clinical effectiveness of the PVS for successful pulmonary artery (PA)/pulmonary vein (PV) transection was evaluated. In addition, the surgeon's stress, device usability, and surgeon satisfaction were measured using multiple questionnaires. Results A total of 167 PAs/PVs were transected with 3 (1.8%) requiring intra-operative intervention. Fourteen of the 50 patients (28.0%) experienced at least one adverse event (AE), among whom 5 (10.0%) suffered from serious AEs. There were no postoperative hemorrhagic complications related to transection of the PA/PV with PVS. Surgeon satisfaction was surveyed by questionnaire after each of the 50 procedures resulting in a 96% reported satisfaction with device usability, specifically related to a low stress load and an increase in work efficiency. Conclusions For VATS lobectomy, the PVS demonstrated a positive surgical effectiveness and value in cognitive and physical distress reduction. Complications following VATS lobectomy to treat NSCLC were generally low and as expected. Intraoperative complications were few and there were no postoperative complications related to the transection of the PA and PV during VATS lobectomy. Favorable results were reported on the surgeon satisfaction questionnaire regarding usability and surgeon stress.

The biosynthesis of the pimelate moiety of biotin in Escherichia coli requires two specialized proteins, BioC and BioH. However, the enzymes that have BioC- or BioH-like activities show remarkable sequence diversity among biotin-producing bacteria. Here, we report that the intracellular rickettsial pathogen Ehrlichia chaffeensis encodes two novel proteins, BioT and BioU, which functionally replace the E. coli BioC and BioH proteins, respectively. The desthiobiotin assays demonstrated that these two proteins make pimeloyl-acyl carrier protein (ACP) from the substrate malonyl-ACP with the aid of the FAS II pathway, through the expected pimeloyl-ACP methyl ester intermediate. BioT and BioU homologues seem restricted to the species of Ehrlichia and its close relative, Anaplasma. Taken together, the synthesis of the biotin precursor in E. chaffeensis appears to be catalyzed by two novel BioC- and BioH-like proteins.

Cellulase is an important enzyme that can be used to breakdown lignocellulose into glucose. Microbulbifer hydrolyticus IRE-31(ATCC 700072) is a kind of marine bacterium, which could grow in high salinity medium and has fast-strong growth ability. In this study, a novel strain was screened from Microbulbifer hydrolyticus IRE-31 through mutations to produce cellulase. The effect of different carbon sources on the growth as well as on the production of cellulase of the new strain was studied. Carboxymethyl-cellulase (CMCase) activity selected to represent cellulase was proven to be effectively promoted while xylose, galactose, and melibiose as well as glucose were used as carbon sources. When xylose and glucose were chosen to be further investigated, 472.57 U/L and 266.01 U/L CMCase activity were obtained from 30 g/L glucose and 10 g/L xylose, respectively. These results clarified the effect of different carbon sources on the production of cellulase, which laid a good foundation for the further research in the production of cellulase by marine bacteria.

PROTEIN PHOSPHATASE4 (PP4) is a highly conserved Ser/Thr protein phosphatase found in yeast, plants, and animals. The composition and functions of PP4 in plants are poorly understood. Here, we uncovered the complexity of PP4 composition and function in Arabidopsis (Arabidopsis thaliana) and identified the composition of one form of PP4 containing the regulatory subunit PP4R3A. We show that PP4R3A, together with one of two redundant catalytic subunit genes, PROTEIN PHOSPHATASE X (PPX)1 and PPX2, promotes the biogenesis of microRNAs (miRNAs). PP4R3A is a chromatin-associated protein that interacts with RNA polymerase II and recruits it to the promoters of miRNA-encoding (MIR) genes to promote their transcription. PP4R3A likely also promotes the cotranscriptional processing of miRNA precursors, because it recruits the microprocessor component HYPONASTIC LEAVES1 to MIR genes and to nuclear dicing bodies. Finally, we show that hundreds of introns exhibit splicing defects in pp4r3a mutants. Together, this study reveals roles for Arabidopsis PP4 in transcription and nuclear RNA metabolism.

Hypoxia is known to be commonly present in breast tumor microenvironments. Stem-like cells that repopulate breast tumors, termed tumor-repopulating cells (TRC), thrive under hypoxic conditions, but the underlying mechanism remains unclear. Here, we show that hypoxia promotes the growth of breast TRCs through metabolic reprogramming. Hypoxia mobilized transcription factors HIF1α and FoxO1 and induced epigenetic reprogramming to upregulate cytosolic phosphoenolpyruvate carboxykinase (PCK1), a key enzyme that initiates gluconeogenesis. PCK1 subsequently triggered retrograde carbon flow from gluconeogenesis to glycogenesis, glycogenolysis, and the pentose phosphate pathway. The resultant NADPH facilitated reduced glutathione production, leading to a moderate increase of reactive oxygen species that stimulated hypoxic breast TRC growth. Notably, this metabolic mechanism was absent in differentiated breast tumor cells. Targeting PCK1 synergized with paclitaxel to reduce the growth of triple-negative breast cancer (TNBC). These findings uncover an altered glycogen metabolic program in breast cancer, providing potential metabolic strategies to target hypoxic breast TRCs and TNBC. Significance: Hypoxic breast cancer cells trigger self-growth through PCK1-mediated glycogen metabolism reprogramming that leads to NADPH production to maintain a moderate ROS level.

Helicobacter pylori is a major global pathogen and has been implicated in gastritis, peptic ulcer, and gastric carcinoma. The efficacy of the extensive therapy of H. pylori infection with antibiotics is compromised by the development of drug resistance and toxicity toward human gut microbiota, which urgently demands novel and selective antibacterial strategies. The present study was mainly performed to assess the in vitro and in vivo effects of a natural herbal compound, dihydrotanshinone I (DHT), against standard and clinical H. pylori strains. DHT demonstrated effective antibacterial activity against H. pylori in vitro (MIC50/90, 0.25/0.5 μg/ml), with no development of resistance during continuous serial passaging. Time-kill curves showed strong time-dependent bactericidal activity for DHT. Also, DHT eliminated preformed biofilms and killed biofilm-encased H. pylori cells more efficiently than the conventional antibiotic metronidazole. In mouse models of multidrug-resistant H. pylori infection, dual therapy with DHT and omeprazole showed in vivo killing efficacy superior to that of the standard triple-therapy approach. Moreover, DHT treatment induces negligible toxicity against normal tissues and exhibits a relatively good safety index. These results suggest that DHT could be suitable for use as an anti-H. pylori agent in combination with a proton pump inhibitor to eradicate multidrug-resistant H. pylori.

Antibiotic resistance in Helicobacter pylori has been growing worldwide with current treatment regimens. Development of new compounds for treatment of H. pylori infections is urgently required to achieve a successful eradication therapy in the future. Armeniaspirols, a novel class of natural products isolated from Streptomyces armeniacus, have been previously identified as antibacterial agents against Gram-positive pathogens. In this study, we found that armeniaspirol A (ARM1) exhibited potent antibacterial activity against H. pylori, including multidrug-resistant strains, with MIC range values of 4-16 μg ml-1 . The underlying mechanism of action of ARM1 against H. pylori involved the disruption of bacterial cell membranes. Also, ARM1 inhibited biofilm formation, eliminated preformed biofilms and killed biofilm-encased H. pylori in a dose-dependent manner. In a mouse model of multidrug-resistant H. pylori infection, dual therapy with ARM1 and omeprazole showed efficient in vivo killing efficacy comparable to the standard triple therapy, and induced negligible toxicity against normal tissues. Moreover, at acidic pH 2.5, ARM1 exhibited a much more potent anti-H. pylori activity than metronidazole. Thus, these findings demonstrated that ARM1 is a novel potent anti-H. pylori agent, which can be developed as a promising drug lead for treatment of H. pylori infections.

Additional file 2: Table S1. Synonymous (dN) and nonsynonymous (dS) substitution rates estimated by PAML. Table S2. The relative expression ratio of the MEP-pathway genes in represented species. Table S3. The Codon Adaption Index of the MEP-pathway genes.

Background: Women with diffuse adenomyosis frequently have severe symptoms, including dysmenorrhea and infertility. Though it is beneficial for long-term health if the uterus can be preserved, there is no prospective investigations to assess lesion removal, symptom alleviation and even gestations after uterus-preservative surgery. Methods: We applied self-optimized PUSH surgery for 146 patients with diffuse adenomyosis and conducted 2 to 11-year follow-up for 114 patients. We detailed procedures in uterine incision , excision of the lesion, uterus reconstruction, solutions for removing giant lesions, and assessed the clinical efficacy and fertility in follow-up. Findings: The average size and volume of the patients’ uterus decreased significantly after PUSH surgery. Operation-related complications occurred in 2.7% (4/146) of the patients. Among 114 patients who had been investigated for ≥2 years, 107 (93.9%, analytic group) had detailed records that was needed to evaluate the long-term outcomes. In analytic group, 5 cases recurred and 95.3% (102/107) of the patients had normal uterine size two years after surgery. The visual analogue scale (VAS) for dysmenorrhea significantly decreased in 102 cases, from 9.35±1.25 to 0.34±0.76 and 0.56±1.06 in the first and second year after surgery, respectively. Additionally, 97.7% of the 86 patients with severe menorrhagia had their symptoms ameliorated in the second year of follow-up. Of the 23 patients who desired for pregnancy and stopped contraception for ≥2 years, 13 (13/23,57%) individuals conceived with17 pregnancies. Twelve patients had 15 intrauterine pregnancies and 7 patients had 8 living fetus. No uterine rupture or severe complications were found in all gestations. Interpretation: This study demonstrate good performance of PUSH surgery in removing uterine lesions, ameliorating menorrhagia and preserving fertility for women with diffuse adenomyosis. Funding: Shenzhen High-level Hospital Construction Fund(YBH2019-260)，Shenzhen Key Medical Discipline Construction Fund(No.SZXK027), Sanming Project of Medicine in Shenzhen(No.SZSM202011016), Scientific Research Foundation of PEKING UNIVERSITY SHENZHEN HOSPITAL(No.KYQD2021075). Declaration of Interest: The authors declare that they have no competing interests. Ethical Approval: The research protocol was approved by the Ethics Committee of Peking University Shenzhen Hospital (IRB approval number: 20190729).

Additional file 1: Figure S1. Phylogenetic tree of DXS. Figure S2. Phylogenetic tree of MDS. Figure S3. Phylogenetic tree of DXR. Figure S4. Phylogenetic tree of HDR. Figure S5. Phylogenetic tree of CMS. Figure S6. Phylogenetic tree of CMK. Figure S7. Phylogenetic tree of HDS.

The flue gas flow in gas turbines is an important parameter for determining the accurate capacity of waste heat boilers for recovering the waste heat released by the flue gas of gas turbines in offshore oil production facilities. The theory of flue gas composition ratio pertaining to boiler combustion is applied to calculate the flue gas flow in gas turbines. The given gas fuel composition, fuel characteristic coefficient and tested dry combustion products of gas fuel are used to calculate the excess air factor α using the theory of flue gas composition ratio; thus, the flue gas flow of gas turbines can be obtained. The results show that the value of α of a gas turbine ranges from 7 to 3 with different loads and is significantly greater than the α value of boilers, which is obtained during the burning of gas fuel; moreover, the calculation mode of O d 2 while considering β coincides more with the actual operation data of gas turbines. This mode is useful for the engineering application of recovering the waste heat released by the flue gas of gas turbines and provides an important guidance to improve the waste heat recovery process in gas turbines.

The phosphopantetheinyl transferases (PPTases) catalyze the post-translational modification of carrier proteins (CPs) from fatty acid synthases (FASs) in primary metabolism and from polyketide synthases (PKSs) and non-ribosomal polypeptide synthases (NRPSs) in secondary metabolism. Based on the conserved sequence motifs and substrate specificities, two types (AcpS-type and Sfp-type) of PPTases have been identified in prokaryotes. We present here that Porphyromonas gingivalis, the keystone pathogen in chronic periodontitis, harbors merely one PPTase, namely PptP. Complementation and gene deletion experiments clearly show that PptP can replace the function of Escherichia coli AcpS and is essential for the growth of P. gingivalis. Purified PptP transfers the 4-phosphopantetheine moiety of CoA to inactive apo-acyl carrier protein (ACP) to form holo-ACP, which functions as an active carrier of the acyl intermediates of fatty acid synthesis. Moreover, PptP exhibits broad substrate specificity, modifying all ACP substrates tested and catalyzing the transfer of coenzyme A (CoA) derivatives. The lack of sequence alignment with known PPTases together with phylogenetic analyses revealed PptP as a new class of PPTases. Identification of the new PPTase gene pptP exclusive in Porphyromonas species reveals a potential target for treating P. gingivalis infections.

Nuclear factor erythroid-2–related factor-2 (NFE2L2/Nrf2) is a transcription factor that regulates the expression of antioxidant genes. Both Kelch-like ECH-associated protein 1 (Keap1) mutations and Nrf2 mutations contribute to the activation of Nrf2 in non-small cell lung cancer (NSCLC). Nrf2 activity is associated with poor prognosis in NSCLC. Metabolic reprogramming represents a cancer hallmark. Increasing studies reveal that Nrf2 activation promotes metabolic reprogramming in cancer. In this review, we discuss the underlying mechanisms of Nrf2-mediated metabolic reprogramming and elucidate its role in NSCLC. Inhibition of Nrf2 can alter metabolic processes, thus suppress tumor growth, prevent metastasis, and increase sensitivity to chemotherapy in NSCLC. In conclusion, Nrf2 may serve as a therapeutic target for the treatment of NSCLC.

BackgroundIsoprenoids are amongst the most ancient and essential class of metabolites produced in all organisms, either via mevalonate (MVA)-and/or methylerythritol dicyclophosphate (MEP)-pathways. The MEP-pathway is present in all plastid-bearing organisms and most eubacteria. However, no comprehensive study reveals the origination and evolutionary characteristics of MEP-pathway genes in eukaryotes.ResultsHere, detailed bioinformatics analyses of the MEP-pathway provide an in-depth understanding the evolutionary history of this indispensable biochemical route, and offer a basis for the co-existence of the cytosolic MVA- and plastidial MEP-pathway in plants given the established exchange of the end products between the two isoprenoid-biosynthesis pathways. Here, phylogenetic analyses establish the contributions of both cyanobacteria and Chlamydiae sequences to the plant’s MEP-pathway genes. Moreover, Phylogenetic and inter-species syntenic block analyses demonstrate that six of the seven MEP-pathway genes have predominantly remained as single-copy in land plants in spite of multiple whole-genome duplication events (WGDs). Substitution rate and domain studies display the evolutionary conservation of these genes, reinforced by their high expression levels. Distinct phenotypic variation among plants with reduced expression levels of individual MEP-pathway genes confirm the indispensable function of each nuclear-encoded plastid-targeted MEP-pathway enzyme in plant growth and development. ConclusionCollectively, these findings reveal the polyphyletic origin and restrict conservation of MEP-pathway genes, and reinforce the potential function of the individual enzymes beyond production of the isoprenoids intermediates.

BackgroundThe incidence and mortality rate of cholangiocarcinoma (CCA) have been rising globally. Patients with CCA have extremely poor prognosis, partly due to the silent clinical character and hence diagnosed at advantage stage without effective treatments. There is growing evidence showing that aberrant expression of messenger RNAs (mRNAs) and long non-coding RNAs (lncRNAs) are involved in tumorigenesis and development of CCA. It is essential to establish an integrated mRNA-lncRNA signature to improve the ability of prognostic prediction in CCA patients.MethodsWe collected a training dataset of 45 patients from The Cancer Genome Atlas dataset and a validation cohort (GSE107943) of 57 patients from Gene Expression Omnibus. An integrated mRNA-lncRNA risk score was established by a univariate and a multivariate Cox regression analyses. Time-dependent receiver operating characteristic (ROC) analysis was used to evaluate prognostic performance. Moreover, we conducted a correlation analysis between the signature and different clinical characteristics, and preformed weighted gene co-expression network analysis (WGCNA) and functional enrichment analysis to investigate functional roles of the integrated signature.ResultsA total of two mRNAs (CFHR3 and PIWIL4) and two lncRNAs (AC007285.1 and AC134682.1) were identified to construct the integrated signature through a univariate Cox regression (P-value = 1.35E-02) and a multivariable Cox analysis (P-value = 1.12E-02). The ROC curve suggested the integrated mRNA-lncRNA signature possessed a high specificity and sensitivity of prognostic prediction with an area under the curve (AUC) of 0.872 and 0.790 at 1-year and 3-years, respectively. Subsequently, the signature was validated in GSE107943 cohort and combined dataset, and an area under the ROC curve reached up to 0.750 and 0.819 at 1-year. The signature was not only independent from different clinical features (P-value= 1.12E-02), but also outperformed other clinical characteristics as prognostic biomarkers with AUC of 0.781 at 3 years. These molecules in the integrated signature may associated with metabolic-related biological process and lipid metabolism pathway, which was highly involved in CCA carcinogenesis. ConclusionThese results showed that the integrated mRNA-lncRNA signature had an independent prognostic value for risk stratification, and further facilitated personalized treatment for CCA patients.

Due to highly maneuverable vessels, dense channels and strong clutter, intermittent track segments are common in large-scale marine monitoring. To solve this problem, we propose a long-term continuous tracking method for high frequency surface wave radar (HFSWR) based on an adaptive filtering algorithm combined with an extreme learning machine (ELM). Via analyzing the features of the vessels and tracks and lots of simulations, we select average velocity, average acceleration, average heading angle, average curvature, ratio of the arc length to the chord length, and wavelet coefficient to compose the multidimensional feature vector to train and test the ELM. Field experiment results show that the proposed method has better performance than conventional algorithms with the correct association probability of 94.7%.

Helicobacter pylori is a major global pathogen and has been implicated in gastritis, peptic ulcer, and gastric carcinoma. The efficacy of the extensive therapy of H. pylori infection with antibiotics is compromised by the development of drug resistance and toxicity toward human gut microbiota, which urgently demands novel and selective antibacterial strategies. The present study was mainly performed to assess the in vitro and in vivo effects of a natural herbal compound, dihydrotanshinone I (DHT), against standard and clinical H. pylori strains. DHT demonstrated effective antibacterial activity against H. pylori in vitro (MIC(50/90), 0.25/0.5 μg/ml), with no development of resistance during continuous serial passaging. Time-kill curves showed strong time-dependent bactericidal activity for DHT. Also, DHT eliminated preformed biofilms and killed biofilm-encased H. pylori cells more efficiently than the conventional antibiotic metronidazole. In mouse models of multidrug-resistant H. pylori infection, dual therapy with DHT and omeprazole showed in vivo killing efficacy superior to that of the standard triple-therapy approach. Moreover, DHT treatment induces negligible toxicity against normal tissues and exhibits a relatively good safety index. These results suggest that DHT could be suitable for use as an anti-H. pylori agent in combination with a proton pump inhibitor to eradicate multidrug-resistant H. pylori.

Dysbiosis of gut microbiota has been implicated in the pathogenesis of hypertension. A definite relationship between gut microbiota and hypertension remains intriguing. Here, we show that the Sacha inchi (Plukenetia volubilis L.) shell extract (SISE) intervention significantly reduced systolic blood pressures in spontaneous hypertensive rats (SHR), attenuated the oxidative damage and modulated plasma calcium homeostasis and left ventricular hypertrophy in both SHR and high-salt diet Wistar-Kyoto rats. SISE reshaped the gut microbiome and metabolome, particularly by improving the prevalence of Roseburia and dihydrofolic acid levels in the gut. Transcriptome analyses showed that the protective effects of SISE were accompanied by the modulation of renal molecular pathways, beneficial for cardiovascular functions such as the L-type voltage-dependent calcium channel (LTCC), a key regulator of calcium signaling. Overall, the results have shown that dietary SISE can alleviate hypertension regulating the gut microbiota, and Ca2+ signaling might be a potential target for spontaneous hypertension.

Background Congenital anatomic abnormalities of fallopian tubes and ovaries are rarely reported. Herein, we describe four cases of abnormal ovarian descent during laparoscopic surgery with abnormal anatomy of fallopian tube, without abnormal uterine development and urinary system abnormalities. These cases are analyzed by their clinical features and their effects on reproductive function. Case presentation For patients with undescended ovary, the upper part of the unilateral/bilateral ovary decline was much higher than that of the common iliac vessel bifurcation, and the fallopian tube on the same side opened in the para-colonic sulcus. Among four patients, two cases had primary infertility, one case had tubal pregnancy rupture and bleeding, and one case had uterine leiomyoma. The development of uterus was normal in all the four patients, and there was no abnormal development of urinary system. During the infertility examination, hysterosalpingography (HSG) suggestion of oviduct lift was a sign for possible undescended ovarian. The pelvic B-ultrasound examination was limited to discover ovarian maldescent. Conclusion Laparoscopy was the gold standard for the diagnosis of poor ovarian decline. When B-ultrasound prompts one-sided or double-sided "ovary display is unclear" or detects "shrinking small ovaries", it might be ovarian maldescent. When there is periodic post-sacral spinal pain, MRI positioning or HSG can be used for diagnosis.