Construction of a New Agrobacterium tumefaciens-Mediated Transformation System based on a Dual Auxotrophic Approach in Cordyceps militaris.

Cordyceps militaris is a significant edible fungus that produces a variety of bioactive compounds. We have previously established a uridine/uracil auxotrophic mutant and a corresponding Agrobacterium tumefaciens-mediated transformation (ATMT) system for genetic characterization in C. militaris using pyrG as a screening marker. In this study, we constructed an ATMT system based on a dual pyrG and hisB auxotrophic mutant of C. militaris. Using the uridine/uracil auxotrophic mutant as the background and pyrG as a selection marker, the hisB gene encoding imidazole glycerophosphate dehydratase, required for histidine biosynthesis, was knocked out by homologous recombination to construct a histidine auxotrophic C. militaris mutant. Then, pyrG in the histidine auxotrophic mutant was deleted to construct a ΔpyrG ΔhisB dual auxotrophic mutant. Further, we established an ATMT transformation system based on the dual auxotrophic C. militaris by using GFP and DsRed as reporter genes. Finally, to demonstrate the application of this dual transformation system for studies of gene function, knock out and complementation of the photoreceptor gene CmWC-1 in the dual auxotrophic C. militaris were performed. The newly constructed ATMT system with histidine and uridine/uracil auxotrophic markers provides a promising tool for genetic modifications in the medicinal fungus C. militaris.

Poor prognosis and risk factors of nonoperative treatment hip fracture patients with end-stage renal disease.

This study aims to summarize the prognosis and risk factors of nonoperative treatment patients of hip fracture with end-stage renal disease (ESRD). A total of 113 individuals of hip fracture with nonoperative treatment were retrospectively enrolled, 38 patients with ESRD were defined as the observation group, and the other 75 patients without ESRD were served as the control group. The difference in 30-day and 1-year mortality rate between the 2 groups was compared. The effects of risk factors on survival were estimated by the Cox proportional-hazards model. The survival difference was estimated by the method of Kaplan-Meier. In the subsequent subgroup analysis of the observation group, as before, the Cox proportional-hazards model and the Kaplan-Meier method were used. The 1-year mortality rate of the observation group was considerably higher than that of the control group, which was 86.84% and 32.0%, respectively (P < .005). For nonoperative treatment hip fracture subjects, ESRD, age ≥ 82, BMI (BMI) < 20, high Charlson Comorbidity Index (CCI) and low Barthel Index (BI) were associated with a low survival curve (P < .05). In the subsequent multivariable subgroup analyses, for nonoperative treatment hip fracture subjects with ESRD, high CCI and low BI were also the independent risk factors of mortality. For nonoperative treatment hip fracture subjects, ESRD, old age, low BMI, high CCI and low BI were the independent risk factors of mortality. For hip fracture subjects with ESRD, nonoperative treatment was associated with excess high 1-year mortality rate, especially for patients with high CCI and low BI.

KPT330 promotes the sensitivity of glioblastoma to olaparib by retaining SQSTM1 in the nucleus and disrupting lysosomal function.

ABBREVIATIONS: AO: acridine orange; ATM: ATM serine/threonine kinase; CHEK1: checkpoint kinase 1; CHEK2: checkpoint kinase 2; CI: combination index; DMSO: dimethyl sulfoxide; DSBs: double-strand breaks; GBM: glioblastoma; HR: homologous recombination; H2AX: H2A.X variant histone; IHC: immunohistochemistry; LAPTM4B: lysosomal protein transmembrane 4 beta; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; PARP: poly(ADP-ribose) polymerase; RAD51: RAD51 recombinase; SQSTM1: sequestosome 1; SSBs: single-strand breaks; RNF168: ring finger protein 168; XPO1: exportin 1.

Research and experiment on the trenching performance of orchard trenching device.

Aiming at the mismatch between the cutter combination of the furrowing device and the operating parameters, and resulting in low quality of furrowing and other problems, the theoretical analysis of the furrowing cutting operation is carried out and the influence law of the furrowing parameters on the trajectory, performance and quality of the furrowing movement is obtained. The influence of trenching parameters on trenching trajectory, performance and quality was obtained. The response surface method was applied to design and carry out field experiments. With the increase of cutter head speed and forward speed, the width and depth of trenching and the thickness of floating soil at the bottom of trenching decreased first and then increased, while the operation power consumption presented the increasing trend gradually. The optimization model of trenching performance quality was constructed to obtain the optimal parameter combination of influencing factors. Field experiments were carried out to verify the optimization results. The optimisation results were verified through field tests, which showed that the average depth of furrowing was 472 mm, the width was 332 mm, the thickness of soil return was 134 mm, and the operating power consumption was 19.95 kW. The results showed that the average depth of furrowing was 472 mm, the width was 332 mm, and the thickness of soil return was 134 mm. The optimization model could meet the operation quality indexes, and provide a theoretical basis for the design of the disc subsection cutting trenching device to select the operation parameter combination required by low power consumption and deep trenching.

Ezrin expression in female reproductive tissues: A review of regulation and pathophysiological implications.

Ezrin, a plasma membrane-microfilament linker, is a cytoskeletal organizer involved in many cellular activities by binding to the membrane protein-ezrin-cytoskeletal protein complex and regulating downstream signal transduction. Increasing evidence demonstrates that ezrin plays an important role in regulating cell polarity, proliferation and invasion. In this study, we analyzed the effects of ezrin on oocytes, follicle development, embryo development and embryo implantation. We reviewed the recent studies on the modalities of ezrin regulation and its involvement in the biological processes of female reproductive physiology and summarized the current research advances in ezrin inhibitors. These studies will provide new strategies and insights for the treatment of diseases.

Corrigendum: Anesthesia management of patients undergoing off-pump coronary artery bypass grafting: A retrospective study of single center.

[This corrects the article DOI: 10.3389/fsurg.2022.1067750.].

Transformation of hydroxylamine to nitrosated and nitrated products during advanced oxidation process.

Recently, hydroxylamine (HAm) was introduced to drive advanced oxidation processes (AOPs) for removing organic contaminants. However, we found that HAm-driven Cu(II)/peroxymonosulfate oxidation of phenol produced p-nitrosophenol, 2-nitrophenol and 4-nitrophenol. The total nitro(so) products accounted for approximately 25.0 % of the phenol transformation at certain condition. SO4•- and •OH were identified as the primary and second significant oxidants, respectively. Reactive nitrogen species (RNS) were involved in phenol transformation. The pathway and mechanism of HAm transformation in HAm-driven transition metal ion-catalyzed AOPs were proposed for the first time in this study. The product of HAm via twice single-electron oxidation by Cu(II) is nitroxyl (HNO/NO-), which is a critical oxidation intermediate of HAm. Further oxidation of HNO by SO4•- or •OH is the initial step in propagating radical chain reactions, leading to nitric oxide radical (•NO) and nitrogen dioxide radical (•NO2) as the primary RNS. HAm is a critical intermediate in natural nitrogen cycle, suggesting that HAm can drive the oxidation processes of pollutants in natural environments. Nitro(so) products will be readily produced when AOPs are applied for ecological remediation. This study highlights the formation of toxic nitrosated and nitrated products in HAm-driven AOPs, and the requirement of risk assessments to evaluate the possible health and ecological impacts.

Levosimendan administration is not associated with increased risk of bleeding and blood transfusion requirement in patients undergoing off-pump coronary artery bypass grafting: a retrospective study from single center.

BACKGROUND: Levosimendan (LEVO) is a positive inotropic drug which could increase myocardial contractility and reduce the mortality rate in cardiac surgical patients. However, Whether LEVO is associated with postoperative bleeding and blood transfusion in cardiac surgical patients is controversial. Therefore, the current study was designed to investigate the impact of LEVO administration on bleeding and blood transfusion requirement in off-pump coronary artery bypass grafting (OPCAB) patients. METHODS: In a retrospective analysis, a total of 292 patients, aged 40-87 years, undergoing elective OPCAB between January 2019 and July 2019, were divided into LEVO group (n = 151) and Control group (n = 141). Patients in LEVO group continuously received LEVO at a rate of 0.1-0.2 µg kg-1 min-1 after anesthesia induction until 24 hours after OPCAB or patients in Control group received no LEVO. The primary outcome was postoperative chest drainage volume. The secondary outcomes were reoperation for postoperative bleeding, transfusion requirement of red blood cells (RBCs), fresh frozen plasma (FFP) and platelet concentrate (PC), etc. Comparisons of two groups were performed with the Student's t-test or Wilcoxon-Mann-Whitney test. RESULTS: There was no significant difference with respect to chest drainage volume ((956.29 ± 555.45) ml vs (1003.19 ± 572.25) ml, p = 0.478) and the incidence of reoperation for postoperative bleeding (1.32% vs 1.42%, p = 0.945) between LEVO group and Control group. The transfusion incidence and volume of allogeneic RBCs, FFP, and PC were comparable between two groups. CONCLUSIONS: LEVO administration was neither associated with more postoperative blood loss nor increased allogeneic blood transfusion requirement in OPCAB patients.

Lifestyle-related risk factors correlated with mental health problems: A longitudinal observational study among 686 male college students in Chongqing, China.

Aim: Public concerns over the mental health problems of college students are rising. Previous research show that female tend to suffer more from mental health problems than males, with few studies focusing on males. This study sought to explore the association of lifestyle-related risk factors with the prevalence of mental health problems among male college students in China. Methods: The lifestyle information and mental health status of 686 male college students from Chongqing, China, were assessed in 2014, and 582 of them were followed up a year later. Participants completed a questionnaire assessing demographic and lifestyle factors which include sleep quality, computer usage, sedentariness, physical activity, smoking, current alcohol, coke, coffee, and milk tea drinking, and current tea/fried food/baked food consumption. Mental health problems were measured using the Depression Anxiety Stress Scale-21 (DASS-21). Results: Univariate analyses indicated that age, sleep latency, sleep duration, computer usage time, milk tea drinking, and fried food consumption were potential risk factors for mental health problems (p's < 0.05). Multivariate analysis further revealed that, either at baseline or during follow-up, participants with (i) more computer usage time were at a higher risk of having depression symptoms (p's < 0.05) and (ii) a higher frequency of fried food consumption were associated with a higher risk of having depression, anxiety, and stress symptoms (p's < 0.05). Additionally, the cross-lagged analysis showed that (i) computer usage time in 2014 is positively correlated with depression status (ß = 0.106, p < 0.05) but not anxiety (ß = 0.047, p > 0.05) and stress (ß = 0.019, p > 0.05) status a year later and (ii) fried food consumption in 2014 is positively correlated with depression (ß = 0.129, p < 0.01), anxiety (ß = 0.168, p < 0.001), and stress (ß = 0.113, p < 0.01) status a year later. Conclusions: Computer usage time and fried food consumption were lifestyle-related risk factors for mental health problems in male college students in Chongqing, China. These results might emphasize further preventive strategies for mental health problems, especially in male college students.

Corrigendum: Regulation of microrna-497-targeting AKT2 influences tumor growth and chemoresistance to cisplatin in lung cancer.

[This corrects the article DOI: 10.3389/fcell.2020.00840.].

The E3 ubiquitin ligase HUWE1 acts through the N-Myc-DLL1-NOTCH1 signaling axis to suppress glioblastoma progression.

BACKGROUND: Elucidation of the post-transcriptional modification has led to novel strategies to treat intractable tumors, especially glioblastoma (GBM). The ubiquitin-proteasome system (UPS) mediates a reversible, stringent and stepwise post-translational modification which is closely associated with malignant processes of GBM. To this end, developing novel therapeutic approaches to target the UPS may contribute to the treatment of this disease. This study aimed to screen the vital and aberrantly regulated component of the UPS in GBM. Based on the molecular identification, functional characterization, and mechanism investigation, we sought to elaborate a novel therapeutic strategy to target this vital factor to combat GBM. METHODS: We combined glioma datasets and human patient samples to screen and identify aberrantly regulated E3 ubiquitin ligase. Multidimensional database analysis and molecular and functional experiments in vivo and in vitro were used to evaluate the roles of HECT, UBA and WWE domain-containing E3 ubiquitin ligase 1 (HUWE1) in GBM. dCas9 synergistic activation mediator system and recombinant adeno-associated virus (rAAV) were used to endogenously overexpress full-length HUWE1 in vitro and in glioma orthotopic xenografts. RESULTS: Low expression of HUWE1 was closely associated with worse prognosis of GBM patients. The ubiquitination and subsequent degradation of N-Myc mediated by HUWE1, leading to the inactivation of downstream Delta-like 1 (DLL1)-NOTCH1 signaling pathways, inhibited the proliferation, invasion, and migration of GBM cells in vitro and in vivo. A rAAV dual-vector system for packaging and delivery of dCas9-VP64 was used to augment endogenous HUWE1 expression in vivo and showed an antitumor activity in glioma orthotopic xenografts. CONCLUSIONS: The E3 ubiquitin ligase HUWE1 acts through the N-Myc-DLL1-NOTCH1 signaling axis to suppress GBM progression. Antitumor activity of rAAV dual-vector delivering dCas9-HUWE1 system uncovers a promising therapeutic strategy for GBM.

Factors associated with left ventricular diastolic dysfunction in patients with septic shock.

PURPOSE: To investigate risk factors associated with left ventricular diastolic dysfunction (LVDD) of patients with septic shock. MATERIALS AND METHODS: Patients with septic shock concomitant with or without LVDD were retrospectively enrolled and divided into the LVDD group (n = 17) and control without LVDD (n = 85). The clinical and ultrasound data were analyzed. RESULTS: A significant (P < 0.05) difference existed between the two groups in serum creatinine, APACHE II score, serum glucose, triglyceride, BUN, FT4, LAVI, mitral E, average e', E/average e', septal e', septal e'/septal s', E/septal e', lateral s', lateral e', and E/lateral e'. LAVI > 37 mL/m2, septal e' < 7 cm/s (OR 11.04, 95% CI 3.38-36.05), septal e'/septal s' < 0.8 (OR 4.09, 95% CI 1.37-12.25), E/septal e' > 15 (OR 22.86, 95% CI 6.09-85.79), lateral e' < 8 cm/s (OR 9.16, 95% CI 2.70-31.07), E/lateral e' > 13 (OR 52, 95% CI 11.99- 225.55), lateral s' < 10 (OR 3.36, 95% CI 1.13-9.99), average e' > 10, E/average e' > 10 (OR 9.53, 95% CI 2.49-36.46), APACHE II score > 16 (OR 3.33, 95% CI 1.00-11.03), SOFA > 5 (or 3.43, 95% CI 1.11-10.60), BUN > 12 mmol/L (OR 3.37, 95% CI 1.15-9.87), serum creatinine > 146 µmol/L (OR 5.08, 95% CI 1.69-15.23), serum glucose > 8 mmol/L (OR 3.36, 95% CI 1.09-10.40), and triglyceride > 1.8 mmol/L were significant (P < 0.05) risk factors for LVDD. LAVI > 37 ml/m2, lateral e' < 8 cm/s, E/lateral e' > 13, and SOFA > 5 were significant (P < 0.05) independent risk factors for LVDD. ROC curve analysis demonstrated that the cut-off value and AUC were 37.09 mL/m2 and 0.85 for LAVI, 8.00 cm/s and 0.89 for lateral e', 12.86 and 0.82 for E/lateral e', and 5.00 and 0.69 for SOFA, respectively. CONCLUSION: Left atrial volume index, mitral lateral e', E/lateral e', and SOFA score are significant independent risk factors for predicting left ventricular diastolic dysfunction in patients with septic shock.

ASCL2 Maintains Stemness Phenotype through ATG9B and Sensitizes Gliomas to Autophagy Inhibitor.

Autophagy is a highly conserved process that is vital for tumor progression and treatment response. Although autophagy is proposed to maintain the stemness phenotype in adult diffuse glioma, the molecular basis of the link between autophagy and stemness is poorly understood, which makes it impossible to effectively screen for the population that will benefit from autophagy-targeted treatment. Here, ATG9B as essential for self-renewal capacity and tumor-propagation potential is identified. Notably, ASCL2 transcriptionally regulates the expression of ATG9B to maintain stemness properties. The ASCL2-ATG9B axis is an independent prognostic biomarker and indicator of autophagic activity. Furthermore, the highly effective blood-brain barrier (BBB)-permeable autophagy inhibitor ROC-325, which can significantly inhibit the progression of ASCL2-ATG9B axisHigh gliomas as a single agent is investigated. These data demonstrate that a new ASCL2-ATG9B signaling axis is crucial for maintaining the stemness phenotype and tumor progression, revealing a potential autophagy inhibition strategy for adult diffuse gliomas.

C-H Direct Arylation: A Robust Tool to Tailor the π-Conjugation Lengths of Non-Fullerene Acceptors.

Facile synthesis without involvement of toxic reagents is of great significance in the practical application of photovoltaic materials. In this work, four acceptor-donor-acceptor (A-D-A) type unfused-ring acceptors (UFRAs) with stepwise extension in π-conjugation, i. e., CPFB-IC-n (n=1-4), involving cyclopentadithiophene (CPDT) and 1,4-difluorobenzene (DFB) as cores, are facilely synthesized by an atom-, step-economic and labor-saving method through direct arylation of C-H bond (DACH). Among them, CPFB-IC-4 has the longest conjugation lengths among the molecular UFRA ever reported. The dependence of optoelectronic properties and photovoltaic performances of CPFB-IC-n (n=1-4) on conjugation length were systematically investigated. CPFB-IC-2 with near zero highest occupied molecular orbital (HOMO) offsets (ΔEHOMO =0.06 eV) achieves the highest power conversion efficiency (PCE), due to the significantly enhanced open voltage (VOC ) and short current (JSC ) caused by the balanced frontier molecular orbitals (FMOs) and complementary light absorption. Our work demonstrates that the optical properties and FMOs of UFRAs can be finely tuned by the stepwise elongation of conjugation lengths. Meanwhile, DACH coupling as a powerful tool here established will be a promising candidate for synthesizing high-performance oligomeric UFRAs.

Multi-View Random-Walk Graph Regularization Low-Rank Representation for Cancer Clustering and Differentially Expressed Gene Selection.

Cancer genome data generally consists of multiple views from different sources. These views provide different levels of information about gene activity, as well as more comprehensive cancer information. The low-rank representation (LRR) method, as a powerful subspace clustering method, has been extended and applied in cancer data research. Although the multi-view learning methods based on low rank representation have achieved good results in cancer multi-omics analysis because they fully consider the consistency and complementarity between views, these methods have some shortcomings in mining the potential local geometry of data. In view of this, this paper proposes a new method named Multi-view Random-walk Graph regularization Low-Rank Representation (MRGLRR) to comprehensively analyze multi-view genomics data. This method uses multi-view model to find the common centroid of view. By constructing a joint affinity matrix to learn the low-rank subspace representation of multiple sets of data, the hidden information of each view is fully obtained. In addition, this method introduces random walk graph regularization constraint to obtain more accurate similarity between samples. Different from the traditional graph regularization constraint, after constructing the KNN graph, we use the random walk algorithm to obtain the weight matrix. The random walk algorithm can retain more local geometric information and better learn the topological structure of the data. What's more, a feature gene selection strategy suitable for multi-view model is proposed to find more differentially expressed genes with research value. Experimental results show that our method is better than other representative methods in terms of clustering and feature gene selection for cancer multi-omics data.

Anesthesia management of patients undergoing off-pump coronary artery bypass grafting: A retrospective study of single center.

Background: To summarize the current practice of anesthesia management for Chinese patients undergoing off-pump coronary artery bypass (OPCAB) surgery at a large-volume cardiovascular center. Materials and methods: The clinical data of consecutive patients undergoing isolated, primary OPCAB surgery during the period from September 2019 to December 2019 were retrospectively analyzed. Patient characteristics, intraoperative data, and short-term outcomes were extracted from the Hospital Information System and the Anesthesia Information Management System. Results: A total of 255 patients who underwent OPCAB surgery were enrolled in the current study. High-dose opioids and short-acting sedatives were the most commonly administrated anesthetics intraoperatively. Pulmonary arterial catheter insertion is frequently performed in patients with serious coronary heart disease. Goal-directed fluid therapy, a restricted transfusion strategy, and perioperative blood management were routinely used. Rational usages of inotropic and vasoactive agents facilitate hemodynamic stability during the coronary anastomosis procedure. Four patients underwent re-exploration for bleeding, but no death was observed. Conclusions: The study introduced the current practice of anesthesia management at the large-volume cardiovascular center, and the short-term outcomes indicated the efficacy and safety of the practice in OPCAB surgery.

Altered Wnt5a expression affects radiosensitivity of non-small cell lung cancer via the Wnt/ß-catenin pathway.

It has been reported that upregulation of wingless-type protein 5a (Wnt5a) is associated with poor prognosis in patients with non-small cell lung cancer (NSCLC). Wnt5a expression is often upregulated in radiation-resistant NSCLC cells. However, the biological functions or molecular mechanisms of radiosensitivity in NSCLC remain unknown. In the present study, MTT assay and flow cytometric analysis were performed to assess the effect of overexpression or knockdown of Wnt5a and/or radiation on the proliferation and apoptosis of NSCLC cells. Furthermore, western blot analysis was performed to detect canonical Wnt signaling (ß-catenin) in H1650 and A549 cells. The results demonstrated that Wnt5a knockdown combined with irradiation inhibited proliferation and induced apoptosis in NSCLC cells compared with Wnt5a knockdown or radiotherapy alone. In addition, the combination of Wnt5a knockdown and irradiation decreased nuclear and increased cytoplasmic ß-catenin expression in H1650 and A549 cells, the effects of which were reversed following overexpression of Wnt5a. The combination of overexpressing Wnt5a and irradiation resulted in significant tumor regression, while ß-catenin knockdown reversed Wnt5a overexpression-induced NSCLC cell proliferation. Taken together, these results suggest that Wnt5a may be involved in the activation of ß-catenin-dependent canonical Wnt signaling, and thus may influence the effectiveness of radiation therapy in NSCLC.

Body Mass Index: An Effective Predictor of Ejection Fraction Improvement in Heart Failure.

Background: Heart failure patients with higher body mass index (BMI) exhibit better clinical outcomes. Therefore, we assessed whether the BMI can predict left ventricular ejection fraction (EF) improvement following heart failure. Methods and Results: We included 184 patients newly diagnosed with dilated cardiomyopathy and reduced EF in our center and who underwent follow-up examination of EF via echocardiography after 6 months. The EF improved at 6 months in 88 participants, who were included in the heart failure with recovered EF (HFrecEF) subgroup. Patients in whom the EF remained reduced were included in the heart failure with persistently reduced EF (persistent HFrEF) subgroup. Our analyses revealed that EF increase correlated with age (r = -0.254, P = 0.001), left ventricular diastolic dimension (LVDD; r = -0.210, P = 0.004), diabetes (P = 0.034), brain natriuretic peptide (r = -0.199, P = 0.007), and BMI grade (P = 0.000). BMI grade was significantly associated with elevated EF after adjustment for other variables (P = 0.001). On multivariable analysis, compared to patients with persistent HFrEF, those with HFrecEF had higher BMI [odds ratio (OR) = 2.342 per one standard deviation increase; P = 0.001] and lower LVDD (OR = 0.466 per one standard deviation increase; P = 0.001). ROC-curve analysis data showed that BMI > 22.66 kg/m2 (sensitivity 84.1%, specificity 59.4%, AUC 0.745, P = 0.000) indicate high probability of EF recovery in 6 months. Conclusions: Our data suggest that higher BMI is strongly correlated with the recovered EF and that BMI is an effective predictor of EF improvement in patients with heart failure and reduced EF.