Whole genome sequencing analysis of seven unknown resistance mechanisms of carbapenem-resistant Klebsiella pneumoniae strains resistance to ceftazidime-avibactam.

AIMS: To investigate alternative resistance mechanisms among seven ceftazidime-avibactam (CZA)-resistant carbapenem-resistant Klebsiella pneumoniae (CRKP) strains lacking common antimicrobial resistance genes (ARGs) using whole genome sequencing. METHODS AND RESULTS: ARG and virulence factors (VFs) were screened using the ARG database CARD and the VF database, respectively, and identified using genomic annotation data with BLAST+. Six strains were ST11 sequence types (STs), and one was ST2123. ST11 strains harbored more ARGs than the ST2123 strains. All seven strains carried multiple ARGs with efflux-mediated antibiotic resistance, including oqxA, oqxB, tet (A), qacEdltal, CRP, H-NS, Kpn-E, F, G, H, acrA, LptD, acrB, acrD, cpxA, mdtB, and mdtC. These efflux-mediated ARGs were identified in most strains and even all strains. Whole genome sequencing revealed that the ST11 strain carried multiple potential prophages, genomic islands, and integrative and conjugative elements, while the ST2123 strain carried an independent potential prophages and a genomic island. CONCLUSIONS: Whole genome sequencing analysis revealed that these seven CZA-resistant CRKP strains lacking common ARGs exhibited efflux-mediated antibiotic resistance-associated ARGs. The main mechanism by which CRKP resists CZA is antibiotic inactivation. Except for tet (A), no ARGs and validation experiments related to efflux were found. This study's results provide a new possibility for the resistance mechanism of CRKP to CZA, and we will verify this conclusion through experiments in the future.

Identification of a broadly neutralizing epitope within Gc protein of Akabane virus using newly prepared neutralizing monoclonal antibodies.

Akabane virus (AKAV) is characterized by abortion, stillbirth, premature birth, and congenital deformities in livestock and is widely distributed throughout Australia, Southeast Asia, East Asia, the Middle East, and Africa. Gc protein is the major neutralizing target of AKAV and is often considered as an immunogen to prepare neutralizing antibodies. In this study, we prepared and characterized three monoclonal antibodies (mAbs), 4D1, 4E6, and 4F12, against the Gc protein of AKAV (TJ2016 strain). Western blot (WB) and indirect immunofluorescence assay (IFA) analysis proved that the mAbs can react with both the truncated recombinant AKAV Gc protein and the natural Gc protein produced in the AKAV-infected cells. Further research demonstrated that these mAbs possess neutralizing activity. We next defined a neutralizing epitope 1134SVQSFDGKL1142 by screening a panel of overlapping peptides spanning the truncated Gc protein (aa991∼1232) using the generated neutralizing mAbs. Bioinformatic analysis shows that the neutralizing epitope is highly conserved across different genotypes of AKAV. The newly produced neutralizing mAbs and the identified neutralizing epitope in this study enrich the antigenic epitope information of the AKAV Gc protein and could have potential applications in the development of antigen and antibody detection systems that are specific to AKAV.

Investigation on the mechanism of androsta-4,6,8,14-tetraene-3,11,16-trione against acute lymphoblastic leukemia.

Steroids are potential anti-leukemia agents, and Epigynum auritum is a Yunnan folk medicine with high levels of androsterone, pregnane, and steroid derivatives. However, the underlying therapeutic mechanism of androsta-4,6,8,14-tetraene-3,11,16-trione (ATT), an androsterone isolated from Epigynum auritum, is not yet clear. This study aimed to explore the anti-leukemia mechanism of ATT using molecular biology, network pharmacology, and molecular docking technology. The cell viability results showed that ATT had an anti-proliferation effect in acute lymphoblastic leukemia cells (CEM/C1, MOLT-4, Jurkat, BALL-1, Nalm-6, and RS4;11). Further studies showed that ATT reduced the mitochondrial membrane potential in B-cell acute lymphoblastic leukemia cell lines (BALL-1, Nalm-6, and RS4;11) and induced cell cycle arrest in MOLT-4 and BALL-1. ATT induced BALL-1 cell apoptosis by activating Caspase 3/7 activity and causing DNA fragmentation. Network pharmacology results suggested that ATT exerts its anti-leukemia activity via the PI3K/Akt signaling pathway. In addition, molecular docking analysis showed that ATT had high scores in docking with PTGS2, NR3C1, and AR. Western blotting results showed that ATT reduced the relative protein level of P-PI3K and P-Akt, thereby increasing the relative level of pro-apoptosis protein Bax and reducing the relative level of anti-apoptosis protein Bcl-2, the apoptosis downstream protein pro-caspase3, and cell proliferation-related proteins (P-GSK3B and CyclinD1). In conclusion, these results demonstrated that ATT could be a potential candidate drug with apoptosis-induction and cell cycle arrest effects for further investigation in acute lymphoblastic leukemia therapy.

Pan-cancer analysis implicates novel insights of lactate metabolism into immunotherapy response prediction and survival prognostication.

BACKGROUND: Immunotherapy has emerged as a potent clinical approach for cancer treatment, but only subsets of cancer patients can benefit from it. Targeting lactate metabolism (LM) in tumor cells as a method to potentiate anti-tumor immune responses represents a promising therapeutic strategy. METHODS: Public single-cell RNA-Seq (scRNA-seq) cohorts collected from patients who received immunotherapy were systematically gathered and scrutinized to delineate the association between LM and the immunotherapy response. A novel LM-related signature (LM.SIG) was formulated through an extensive examination of 40 pan-cancer scRNA-seq cohorts. Then, multiple machine learning (ML) algorithms were employed to validate the capacity of LM.SIG for immunotherapy response prediction and survival prognostication based on 8 immunotherapy transcriptomic cohorts and 30 The Cancer Genome Atlas (TCGA) pan-cancer datasets. Moreover, potential targets for immunotherapy were identified based on 17 CRISPR datasets and validated via in vivo and in vitro experiments. RESULTS: The assessment of LM was confirmed to possess a substantial relationship with immunotherapy resistance in 2 immunotherapy scRNA-seq cohorts. Based on large-scale pan-cancer data, there exists a notably adverse correlation between LM.SIG and anti-tumor immunity as well as imbalance infiltration of immune cells, whereas a positive association was observed between LM.SIG and pro-tumorigenic signaling. Utilizing this signature, the ML model predicted immunotherapy response and prognosis with an AUC of 0.73/0.80 in validation sets and 0.70/0.87 in testing sets respectively. Notably, LM.SIG exhibited superior predictive performance across various cancers compared to published signatures. Subsequently, CRISPR screening identified LDHA as a pan-cancer biomarker for estimating immunotherapy response and survival probability which was further validated using immunohistochemistry (IHC) and spatial transcriptomics (ST) datasets. Furthermore, experiments demonstrated that LDHA deficiency in pancreatic cancer elevated the CD8+ T cell antitumor immunity and improved macrophage antitumoral polarization, which in turn enhanced the efficacy of immunotherapy. CONCLUSIONS: We unveiled the tight correlation between LM and resistance to immunotherapy and further established the pan-cancer LM.SIG, holds the potential to emerge as a competitive instrument for the selection of patients suitable for immunotherapy.

Multiple systemic infections caused by Rhodococcus equi: a case report.

Background: Rhodococcus equi is one of the most important causes of zoonotic infections from grazing animals. It poses a particular risk to immunocompromised individuals, including those who are undergoing long-term immunosuppressive therapy. Case presentation: We report a case of Rhodococcus equi infection in a 65-year-old man with a medical history of diabetes, hypertension, and Adult Still's Disease, currently taking long-term hormone therapy. The non-human immunodeficiency virus (HIV)-infected patient had blood, lung tissue, and sputum samples infected with Rhodococcus equi. His condition initially failed to improve despite multiple therapies, including vancomycin and meropenem. Although his symptoms improved after shifting his antibiotics to cover for the causative agent, he did not completely recover upon hospital discharge. Conclusions: In recent years, the number of Rhodococcus equi cases has increased. This report describes a lethal case of Rhodococcus equi infection in a patient without HIV.

IGF2BP2-related modification patterns in pancreatic cancer: A machine learning-driven approach towards personalized treatment.

Pancreatic cancer (PC) is a malignant digestive system tumor with a very poor prognosis. N6-methyladenosine (m6A) is mediated by a variety of readers and participates in important regulatory roles in PC. Based on TCGA_PAAD, ICGC_AU_PAAD, ICGC_CA_PAAD, GSE28735 and GSE62452 datasets, We mapped the multi-omics changes of m6A readers in PC and found that m6A readers, especially IGF2BP family genes, had specific changes and were significantly associated with poor prognosis. An unsupervised consensus clustering algorithm was used to explore the correlation between specific expression patterns of m6A readers in PC and enrichment pathways, tumor immunity and clinical molecular subtypes. Then, the principal component analysis (PCA) algorithm was used to quantify specific expression patterns and screen core genes. Machine learning algorithms such as Bootstrapping and RSF were used to quantify the expression patterns of core genes and construct a prognostic scoring model for PC patients. What's more, pharmacogenomic databases were used to screen sensitive drug targets and small molecule compounds for high-risk PC patients in an all-around and multi-angle way. Our study has not only provided new insights into personalized prognostication approaches, but also thrown light on integrating tailored risk stratification with precision therapy based on IGF2BP2-mediated m6A modification patterns.

Clinical outcomes of endodontic microsurgery in complicated cases with large or through-and-through lesions: a retrospective longitudinal study.

OBJECTIVES: To investigate the clinical outcomes of endodontic microsurgery in complicated cases presenting with large or through-and-through lesions. MATERIALS AND METHODS: We retrospectively collected and analyzed preoperative, intraoperative, and follow-up data from 143 complicated cases that underwent endodontic microsurgery. Clinical outcomes were assessed in terms of tooth survival and surgery success. Cox regression analysis was used to evaluate the survival rate and identify associated risk factors. Additionally, the success rate was compared across different postoperative periods, and potential factors contributing to surgical failure were identified through binary logistic regression. RESULTS: The overall survival and success rates were 93.0% and 91.7%, respectively. The Cox regression model identified four risk factors affecting tooth survival, including apicoectomy of four teeth (HR = 35.488; P = 0.0002), an open apex observed on preoperative radiographs (HR = 6.300; P = 0.025), the performance of guided tissue regeneration technique (HR = 8.846; P = 0.028), and a palatal surgical approach (HR = 8.685; P = 0.030). The success rate demonstrated an initial increase in the early postoperative period (from 0.5 to 2 years; P = 5.8124e-30), followed by stabilization (from 2 to 9 years; P = 0.298). Surgery success rate significantly declined when apicoectomy involved four teeth (OR = 109.412; P = 0.002). CONCLUSIONS: Endodontic microsurgery demonstrates satisfactory outcomes in complicated cases, maintaining a stable success rate after two years. However, tooth survival and surgery success are significantly compromised when apicoectomy involves four teeth. Factors such as guided tissue regeneration, an open apex, and the palatal surgical approach are associated with an increased risk of tooth extraction. CLINICAL RELEVANCE: Despite achieving acceptable outcomes in complicated cases, endodontic microsurgery is adversely affected by the apicoectomy of four teeth.

Analogies between photochemical reactions and ground-state post-transition-state bifurcations shed light on dynamical origins of selectivity.

Revealing the origins of kinetic selectivity is one of the premier tasks of applied theoretical organic chemistry, and for many reactions, doing so involves comparing competing transition states. For some reactions, however, a single transition state leads directly to multiple products, in which case non-statistical dynamic effects influence selectivity control. The selectivity of photochemical reactions-where crossing between excited-state and ground-state surfaces occurs near ground-state transition structures that interconvert competing products-also should be controlled by the momentum of the reacting molecules as they return to the ground state in addition to the shape of the potential energy surfaces involved. Now, using machine-learning-assisted non-adiabatic molecular dynamics and multiconfiguration pair-density functional theory, these factors are examined for a classic photochemical reaction-the deazetization of 2,3-diazabicyclo[2.2.2]oct-2-ene-for which we demonstrate that momentum dominates the selectivity for hexadiene versus [2.2.2] bicyclohexane products.

Comparative pharmacokinetic analysis of sporoderm-broken and sporoderm-removed Ganoderma lucidum spore in rat by using a sensitive plasma UPLC-QqQ-MS method.

Previous studies have found that removing the sporoderm significantly enhanced antitumor and immunoregulatory activities of Ganoderma lucidum spore (GLS) compared with breaking the sporoderm. However, the pharmacokinetics of sporoderm-removed GLS (RGLS) and sporoderm-broken GLS (BGLS) remain elusive. To compare the pharmacokinetic differences between the two products, we developed a UPLC-QqQ MS method for determining nine representative triterpenoid concentrations. Chloramphenicol was used as an internal standard. The samples were separated on a reversed-phase column using acetonitrile-0.1% formic acid and water-0.1% formic acid as mobile phases. Nine triterpenoids were analyzed using multiple reaction monitoring mode. The results showed that the area under the concentration-time curve from dosing to time t of all nine components was increased in RGLS compared with BGLS. And the time to the maximum concentration in BGLS was delayed compared with that of RGLS. These indicated that the absorption of RGLS was better than that of BGLS, and the sporoderm might hinder the absorption of the active components. These results increase our understanding of the bioavailability of BGLS and RGLS and indicate that increased bioavailability is one of the main reasons for the enhanced efficacy of RGLS.

A survey of methods for handling initial state shifts in iterative learning control.

This paper introduces three types of controllers: a PID-type iterative learning controller, an adaptive iterative learning controller, and an optimal iterative learning controller, and reviews the history and research status of initial shifts rectifying algorithms. Initial state shifts have attracted research attention because they affect both the tracking performance and system stability. This study focuses on the current common initial shifts rectifying methods and analyzes the underlying mechanism in detail. To verify the effectiveness of the presented initial shifts rectifying algorithms, we simulated those using ideal first- and second-order systems. Finally, directions for the future development of iterative learning control (ILC) and some challenging topics related to initial shifts rectifying for ILC are presented. This article aims to introduce recent developments and advances in initial shifts rectifying algorithms and discuss the directions for their further exploration.

Effects of Bulk LPSO Phases on Mechanical Properties and Fracture Behavior of As-Extruded Mg-Gd-Y-Zn-Zr Alloys.

Despite the consensus on the constructive effect of LPSO (long-period stacking-ordered) phases, the true effect of bulk LPSO phases on strengthening and toughening in deformed magnesium alloys is still controversial. This article, which introduces the alloys Mg-8Gd-4Y-0.6Zn-0.5Zr, without bulk LPSO phases, and Mg-8Gd-4Y-1.6Zn-0.5Zr, containing bulk LPSO phases, details a systematically comparative analysis conducted to clarify the true contribution of bulk LPSO phases to the properties of as-extruded alloys. The results indicate that bulk LPSO phases significantly improve strength by refining grain sizes remarkably. But contrary to expectations, bulk LPSO phases themselves only provide a small strengthening effect and deteriorate plasticity, ascribed to the poor compatible plastic deformation of bulk LPSO phases. More importantly, this work may offer new insights into the strengthening and toughening of LPSO phases for further research and engineering applications of this series of alloys. Additionally, an example of a design strategy for Mg-Gd-Y-Zn alloys with superior strength and excellent plasticity is proposed at the end of this article.

Machine learning algorithm integrates bulk and single-cell transcriptome sequencing to reveal immune-related personalized therapy prediction features for pancreatic cancer.

Pancreatic cancer (PC) is a digestive malignancy with worse overall survival. Tumor immune environment (TIME) alters the progression and proliferation of various solid tumors. Hence, we aimed to detect the TIME-related classifier to facilitate the personalized treatment of PC. Based on the 1612 immune-related genes (IRGs), we classified patients into Immune_rich and Immune_desert subgroups via consensus clustering. Patients in distinct subtypes exhibited a difference in sensitivity to immune checkpoint blockers (ICB). Next, the immune-related signature (IRS) model was established based on 8 IRGs (SYT12, TNNT1, TRIM46, SMPD3, ANLN, AFF3, CXCL9 and RP1L1) and validated its predictive efficiency in multiple cohorts. RT-qPCR experiments demonstrated the differential expression of 8 IRGs between tumor and normal cell lines. Patients who gained lower IRS score tended to be more sensitive to chemotherapy and immunotherapy, and obtained better overall survival compared to those with higher IRS scores. Moreover, scRNA-seq analysis revealed that fibroblast and ductal cells might affect malignant tumor cells via MIF-(CD74+CD44) and SPP1-CD44 axis. Eventually, we identified eight therapeutic targets and one agent for IRS high patients. Our study screened out the specific regulation pattern of TIME in PC, and shed light on the precise treatment of PC.

Comparison of Thermal Deformation Behavior and Characteristics of Mg-Gd-Y-Zn Alloys with and without Bulk LPSO Phase.

Alloys Mg-8Gd-4Y-0.6Zn-0.5Zr (referred to as 0.6Zn) without the bulk long-period stacking ordered (LPSO) phase and Mg-8Gd-4Y-1.1Zn-0.5Zr (referred to as 1.1Zn) containing the bulk LPSO phase were prepared and a series of hot compression tests were conducted to examine and evaluate the influence of the bulk LPSO phase on the thermal deformation behavior and characteristics of the Mg-Gd-Y-Zn-Zr alloy. The bulk LPSO phase affects the dynamic recrystallization behavior, resulting in differences in flow stress between two alloys under different conditions. Specifically, in the temperature range of 380~460 °C, compression at lower strain rates is beneficial for the LPSO phase to promote dynamic recrystallization, while compression at a high strain rate inhibits the dynamic recrystallization due to the severe deformation of the bulk LPSO phase to release the stress concentration instead. The increase in temperature helps the LPSO promote dynamic recrystallization. As a result, the LPSO phase promotes dynamic recrystallization at all experimental strain rates at 500 °C. Furthermore, the thermal processing maps of the 0.6Zn and 1.1Zn alloys are established, and their optimal processing windows are located at 500 °C/0.001~0.01 s-1 and 500 °C/0.01 s-1, respectively. In addition, the instability zones for the 1.1Zn alloy are much larger than that for the 0.6Zn alloy, which corresponds to the microcracks generated at the interfaces between α-Mg and bulk LPSO phases.

Generation of Stable Cell Lines Expressing Akabane Virus N Protein and Insight into Its Function in Viral Replication.

Akabane virus (AKAV) is a world wide epidemic arbovirus belonging to the Bunyavirales order that predominantly infects livestock and causes severe congenital malformations. The nucleocapsid (N) protein of AKAV possesses multiple important functions in the virus life cycle, and it is an ideal choice for AKAV detection. In this study, we successfully constructed two stable BHK-21 cell lines (C8H2 and F7E5) that constitutively express the AKAV N protein using a lentivirus system combined with puromycin selection. RT-PCR analysis confirmed that the AKAV N gene was integrated into the BHK-21 cell genome and consistently transcribed. Indirect immunofluorescence (IFA) and Western blot (WB) assays proved that both C8H2 and F7E5 cells could react with the AKAV N protein mAb specifically, indicating potential applications in AKAV detection. Furthermore, we analyzed the growth kinetics of AKAV in the C8H2 and F7E5 cell lines and observed temporary inhibition of viral replication at 12, 24 and 36 h postinfection (hpi) compared to BHK-21 cells. Subsequent investigations suggested that the reduced viral replication was linked to the down-regulation of the viral mRNAs (Gc and RdRp). In summary, we have established materials for detecting AKAV and gained new insights into the function of the AKAV N protein.

Comprehensive Analysis of Physiological, Biochemical and Flavor Characteristics Changes in Crucian Carp (Carassius auratus) under Different Concentrations of Eugenol.

Eugenol is a widely used fishery anesthetic. This study investigated the effects of various concentrations of eugenol on blood physiological and biochemical indexes, and muscle flavor, in crucian carp (Carassius auratus). To determine the appropriate concentration of eugenol anesthetic for use in crucian carp transportation and production operations, we evaluated seven anesthesia groups of 20, 30, 40, 50, 60, 70, and 80 mg/L and one control group (without eugenol) to determine the effects on blood physiological and biochemical indexes, and muscle flavor. The red blood cells and platelets of crucian carp decreased significantly (p < 0.05) with eugenol treatment. With increasing eugenol concentration, the white blood cells and hemoglobin did not change significantly, whereas lactate dehydrogenase, alkaline phosphatase, alanine aminotransferase, and aspartate aminotransferase increased significantly (p < 0.05). The content of phosphorus, magnesium, and sodium increased after anesthesia, whereas the content of potassium decreased with increasing eugenol concentration. After anesthesia, the content of albumin and total protein in the serum decreased with increasing eugenol concentration (p < 0.05); triglyceride first increased and subsequently decreased (p < 0.05); blood glucose content first increased and then decreased (p < 0.05); and no significant difference was observed in total cholesterol content (p > 0.05). No significant difference was observed in muscle glycogen and liver glycogen content after eugenol anesthesia (p > 0.05). The eugenol-based anesthesia test did not indicate major liver histomorphological effects, but the very small number of gill sheet edema cases observed requires further study. Analysis of electronic nose data indicated that eugenol treatment affected the flavor of the fish. The anesthesia concentration of 20-80 mg/L had some effect on the physiology and biochemistry of crucian carp, thus providing a reference for the application of eugenol in crucian carp transportation and experimental research.

Study on the Regulation Mechanism of Quality Deterioration Due to Chilling Stress and Dry Exposure during Anhydrous Storage and Transportation of Yesso Scallop Patinopecten yessoensis.

In this paper, the quality change of Yesso scallop (Patinopecten yessoensis) in the process of anhydrous storage and transportation after cold acclimation and induced dormancy was studied, and the regulation mechanism of quality degradation during storage and transportation in the process of gradient chilling stress and drying exposure was further explored. The results show that, when transferred from hydrous to anhydrous states, the breathing pattern of the scallops changed from aerobic to anaerobic. Their gill filaments were altered and their apparent vitality constantly declined, which was reflected by the edge shrinkage of the pallium and the direct proportions of the edge reduction rate and the stimulus response period. After being in the anhydrous state for 4 d, the AEC value dropped to 67.59%. At this time, if they were placed under hydration again, the scallops resumed a good growth state. By proteomics analysis, it was revealed that cold acclimation and dry exposure mainly led to changes in biological functions and pathways, such as mitochondrial inner membrane and ATP hydrolysis activity. In addition, it can be seen from the functional annotation and enrichment analysis of the metabolite KEGG that cold acclimation promoted the purine metabolism of scallops, while dry exposure inhibited the metabolism of saturated fatty acids. In this study, the infrared sensing mode was used for the first time, too, in order to record the heart-rate changes of the scallops during circulation, which shows that non-destructive vitality monitoring of Lamellibranchia is feasible.

Synthesis, anti-leukemia activity, and molecular docking of novel 3,16-androstenedione derivatives.

In this study, we synthesized androsta-4,14-diene-3,16-dione, 12ß-hydroxyandrosta-4,14-diene-3,16-dione, and other 3,16-androstenedione derivatives from commercially available dehydroepiandrosterone as a starting material in 9-13 steps with high yields. The bioactivity of the obtained compounds was evaluated. Compounds 14a and 23a were shown to have high antitumor activity against acute lymphoblastic leukemia cell lines Nalm-6 and BALL-1, respectively. Network pharmacology analysis showed that the anti-leukemia activity of compounds 14a and 23a might be related to the JAK2, ABL1 protein, and PI3K/Akt signaling pathways. The molecular docking of compounds 14a and 23a identified possible active sites, with the lowest docking scores for PTGS2 and MAPK14, respectively. In addition, the absorption, distribution, metabolism, and excretion prediction results revealed the drug-likeness of the two compounds. Therefore, compounds 14a and 23a should be considered anti-leukemia candidates in future studies.

Effects of ceramsite derived from sewage sludge combined with the O3-FeCl3/PAM process on the dewatering of waste-activated sludge and investigation of dewatering mechanisms.

The high water content of waste-activated sludge restricts the subsequent disposal of sludge. The dehydration properties of O3, ferric chloride (FeCl3)/polyacrylamide, and sludge ceramsite sand (SCS) were studied. Simultaneously, the effect of combining the three was investigated to support the deep dehydration of waste-activated sludge. Experimental results showed that with13.42 mg/(g dry solids (DS)) of O3, 109.89 mg/(g DS) of FeCl3, and 100 mesh dosage of 70% DS of sludge ceramsite on weight, the highest sludge net yield was 7.13 kg/(m2·h) and the minimum specific resistance to filtration of sludge cake was 1.02 × 1012 (m/kg). Compared with the compressibility of the raw sludge, the compressibility of the sludge cake decreased by 37.48%. Moreover, the YN (net yield) increased by 73.55%. The results demonstrate that the structure of cracking, flocculation, and hydrophobic framework is the mechanism of sludge dewatering in this combined process. This combined treatment process provides a new perspective for the realization of deep dewatering of sludge and is anticipated to be a successful sludge dehydration method.

Comprehensive machine learning-generated classifier identifies pro-metastatic characteristics and predicts individual treatment in pancreatic cancer: A multicenter cohort study based on super-enhancer profiling.

Rationale: Accumulating evidence illustrated that the reprogramming of the super-enhancers (SEs) landscape could promote the acquisition of metastatic features in pancreatic cancer (PC). Given the anatomy-based TNM staging is limited by the heterogeneous clinical outcomes in treatment, it is of great clinical significance to tailor individual stratification and to develop alternative therapeutic strategies for metastatic PC patients based on SEs. Methods: In our study, ChIP-Seq analysis for H3K27ac was performed in primary pancreatic tumors (PTs) and hepatic metastases (HMs). Bootstrapping and univariate Cox analysis were implemented to screen prognostic HM-acquired, SE-associated genes (HM-SE genes). Then, based on 1705 PC patients from 14 multicenter cohorts, 188 machine-learning (ML) algorithm integrations were utilized to develop a comprehensive super-enhancer-related metastatic (SEMet) classifier. Results: We established a novel SEMet classifier based on 38 prognostic HM-SE genes. Compared to other clinical traits and 33 published signatures, the SEMet classifier possessed robust and powerful performance in predicting prognosis. In addition, patients in the SEMetlow subgroup owned dismal survival rates, more frequent genomic alterations, and more activated cancer immunity cycle as well as better benefits in immunotherapy. Remarkably, there existed a tight correlation between the SEMetlow subgroup and metastatic phenotypes of PC. Among 18 SEMet genes, we demonstrated that E2F7 may promote PC metastasis through the upregulation of TGM2 and DKK1. Finally, after in silico screening of potential compounds targeted SEMet classifier, results revealed that flumethasone could enhance the sensitivity of metastatic PC to routine gemcitabine chemotherapy. Conclusion: Overall, our study provided new insights into personalized treatment approaches in the clinical management of metastatic PC patients.

Effect of Vanillin on the Anaesthesia of Crucian Carp: Effects on Physiological and Biochemical Indices, Pathology, and Volatile Aroma Components.

The anaesthetic effect of vanillin on crucian carp was investigated using different concentrations of vanillin, with a nonvanillin control. The effective concentration range of vanillin anaesthesia was determined from the behavioural characteristics of crucian carp during the anaesthesia onset and recovery phases. Physiological and biochemical indices, and the electronic nose response to the fish muscle, were measured over the range of effectiveanaestheticc concentrations. An increased concentration of vanillin shortened the time taken to achieve deep anaesthesia but increased the recovery time. The levels of white blood cells, red blood cells, haemoglobinn, platelets, alanine aminotransferase, alkaline phosphatase, lactate dehydrogenase, phosphorus, potassium, magnesium, total protein, and serum albumin were lower than the control in the vanillin treatment group. Triglycerides and total cholesterol were not significantly affected. Histology showed no effect of vanillin on the liver, except at 1.00 g/L vanillin. Vanillin resulted in a nondose-responsive effect on the gill tissue, increasing the width and spacing of the gill lamellae. E-Nose analysis of the carp-muscle flavour volatiles was able to distinguish between different vanillin treatment concentrations. GC-IMS identified 40 flavour compounds, including 8 aldehydes, 11 alcohols, 10 ketones, 2 esters, and 1 furan. Vanillin had aanaestheticic effect on crucian carp and these findings provide a theoretical basis for improving the transport and experimental manipulation of crucian carp.