Graphene Oxide Quantum Dots-Preactivated Dental Pulp Stem Cells/GelMA Facilitates Mitophagy-Regulated Bone Regeneration.

Background: In bone tissue engineering (BTE), cell-laden scaffolds offer a promising strategy for repairing bone defects, particularly when host cell regeneration is insufficient due to age or disease. Exogenous stem cell-based BTE requires bioactive factors to activate these cells. Graphene oxide quantum dots (GOQDs), zero-dimensional derivatives of graphene oxide, have emerged as potential osteogenic nanomedicines. However, constructing biological scaffolds with GOQDs and elucidating their biological mechanisms remain critical challenges. Methods: We utilized GOQDs with a particle size of 10 nm, characterized by a surface rich in C-O-H and C-O-C functional groups. We developed a gelatin methacryloyl (GelMA) hydrogel incorporated with GOQDs-treated dental pulp stem cells (DPSCs). These constructs were transplanted into rat calvarial bone defects to estimate the effectiveness of GOQDs-induced DPSCs in repairing bone defects while also investigating the molecular mechanism underlying GOQDs-induced osteogenesis in DPSCs. Results: GOQDs at 5 µg/mL significantly enhanced the osteogenic differentiation of DPSCs without toxicity. The GOQDs-induced DPSCs showed active osteogenic potential in three-dimensional cell culture system. In vivo, transplantation of GOQDs-preactivated DPSCs/GelMA composite effectively facilitated calvarial bone regeneration. Mechanistically, GOQDs stimulated mitophagy flux through the phosphatase-and-tensin homolog-induced putative kinase 1 (PINK1)/Parkin E3 ubiquitin ligase (PRKN) pathway. Notably, inhibiting mitophagy with cyclosporin A prevented the osteogenic activity of GOQDs. Conclusion: This research presents a well-designed bionic GOQDs/DPSCs/GelMA composite scaffold and demonstrated its ability to promote bone regeneration by enhancing mitophagy. These findings highlight the significant potential of this composite for application in BTE and underscore the crucial role of mitophagy in promoting the osteogenic differentiation of GOQDs-induced stem cells.

Longitudinal assessment of quality of life indicators and prognosis in esophageal cancer patients with curative resection.

Background: The relationship between quality of life and survival outcomes in esophageal cancer patients following curative resection is not well established. This study aimed to longitudinally assess quality of life indicators and their association with overall survival (OS) in these patients. Methods: A total of 232 patients were included in the study, and their quality of life was prospectively assessed at different time points using the European Organisation for Research and Treatment of Cancer (EORTC) 30-item core quality of life questionnaire (QLQ-C30) and the disease-specific esophageal module (QLQ-OES18). The scores of QLQ indicators at each time point were summarized, and changes in postoperative assessment were compared with preoperative assessments. The association of deterioration in certain indicators with OS was evaluated at each time point using Cox univariable analysis. Further confirmation of independent variables was carried out using Cox multivariable analysis. Results: The study cohort comprised 62 females (26.7%), and 113 patients (48.7%) aged over 60 years. The median follow-up time was 80 months (range, 8-118 months). At 24 months after discharge, patients reported improvements in role function, fatigue, cognition function, emotional function, social function, insomnia, appetite loss, nausea and vomiting, constipation, financial status, trouble swallowing saliva, and pain related to esophageal cancer. However, physical function, dyspnea, diarrhea, global health status, choking when swallowing, trouble talking, and reflux remained compromised. Multivariable regression analysis revealed deterioration in role function, emotional function, and coughing difficulty at 6 months, and dyspnea, pain, and cognitive function at 24 months post-discharge were identified as independent prognostic factors for OS. Conclusions: Our findings underscore the importance of monitoring quality of life indicators in esophageal cancer patients as they may significantly influence survival outcomes. The identification of specific quality of life indicators as prognostic factors highlights the need for a patient-centered approach in clinical practice to enhance care and potentially improve survival.

Knocking Out of UDP-Glycosyltransferase Gene UGT2B10 via CRISPR/Cas9 in Helicoverpa armigera Reveals Its Function in Detoxification of Insecticides.

The role of insect UDP-glycosyltransferases (UGTs) in the detoxification of insecticides has rarely been reported. A UGT gene UGT2B10 was previously found overexpressed in a fenvalerate-resistant strain of Helicoverpa armigera. Herein, UGT2B10 was cloned, and its involvement in insecticide detoxification was investigated. UGT2B10 was highly expressed in the larvae, mainly in the fat body and midgut. Treatment with UGT inhibitors 5-nitrouracil and sulfinpyrazone significantly enhanced the fenvalerate toxicity. Knocking down UGT2B10 by RNAi significantly increased the larvae mortality by 17.89%. UGT2B10 was further knocked out by CRISPR/Cas9, and a homozygous strain (HD-dUGT2B10) with a C-base deletion at exon 2 was obtained. The sensitivity of HD-dUGT2B10 to fenvalerate, deltamethrin, cyantraniliprole, acetamiprid, and lufenuron increased significantly, with sensitivity index increased 2.523-, 2.544-, 2.250-, 2.473-, and 3.556-fold, respectively. These results suggested that UGT2B10 was involved in the detoxification of H. armigera to insecticides mentioned above, shedding light upon further understanding of the detoxification mechanisms of insecticides by insect UGTs.

Improving the Retention Rate of Fat Grafting by Botulinum Toxin A: A Randomized, Self-controlled, Clinical Trial.

BACKGROUND: Botulinum toxin A (BTX-A) can enhance the efficacy of fat grafting. However, most studies conducted animal experiments, lacked objective clinical data, or were non-randomized controlled trials. Thus, objective evaluation of the clinical effectiveness of BTX-A is still elusive. METHODS: A randomized, self-controlled trial (2020-2022) on 16 patients who underwent bilateral autogenous fat breast augmentation was performed with each patient receiving autologous fat graft and BTX-A on one side and fat graft and equal volume of saline on the other side. All patients were followed. The effects of BTX-A were evaluated objectively by comparing the remaining bilateral fat graft volumes obtained through digital three-dimensional reconstruction. The improvement of each breast appearance and complication were assessed by the physician and patients who were blinded to the treatment. RESULTS: The outcome of fat breast augmentation was evident for both sides at follow-up with no evidence of fat embolism, vascular/nervous injury, infection, and prolonged bruising. The analysis of the three-dimensional reconstruction data and assessments from both physicians and patients showed significant differences in the fat graft retention volume between the BTX-A side and the control side. No significant difference was found in the incidence of complications between the two sides. CONCLUSIONS: Autogenous fat breast augmentation is safe and effective. This study shows that BTX-A can significantly improve the retention rate of fat transplantation, but cannot reduce complications. Trial registration This study was registered prior to patient enrollment (ClinicalTrials.gov identifier:ChiCTR2100054878). LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Engineered Enucleated Mesenchymal Stem Cells Regulating Immune Microenvironment and Promoting Wound Healing.

Persistent excessive inflammation caused by neutrophil and macrophage dysfunction in the wound bed leads to refractory response during wound healing. However, previous studies using cytokines or drugs often suffer from short half-lives and limited targeting, resulting in unsatisfactory therapeutic effects. Herein, the enucleated mesenchymal stem cell is engineered by aptamer bioorthogonal chemistry to modify the cell membrane and mRNA loading in the cell cytoplasm as a novel delivery vector (Cargocyte) with accurate targeting and sustained cytokine secretion. Cargocytes can successfully reduce NETosis by targeting the nuclear chromatin protein DEK protein with aptamers and sustaining interleukin (IL)-4 expression to overcome the challenges associated with the high cost and short half-life of IL-4 protein and significantly prevent the transition of macrophages into the M1 phenotype. Therapeutic effects have been demonstrated in murine and porcine wound models and have powerful potential to improve wound immune microenvironments effectively. Overall, the use of engineered enucleated mesenchymal stem cells as a delivery system may be a promising approach for wound healing.

Marital status impacts survival of stage I non-small-cell lung cancer: a propensity-score matching analysis.

Aim: This population-based analysis aimed to explore the associations among marital status, prognosis and treatment of stage I non-small-cell lung cancer. Materials & methods: The propensity score matching (PSM), logistic regression and Cox proportional hazards model were used in this study. Results: A total of 13,937 patients were included. After PSM, 10579 patients were co-insured. The married were more likely to receive surgical treatment compared with the unmarried patients (OR: 1.841, p < 0.001), and patients who underwent surgery also tended to have better survival (HR: 0.293, p < 0.001). Conclusion: Compared with unmarried patients, a married group with stage I NSCLC had timely treatment and more satisfactory survival. This study highlights the importance of prompt help and care for unmarried patients.

Expert consensus on endodontic therapy for patients with systemic conditions.

The overall health condition of patients significantly affects the diagnosis, treatment, and prognosis of endodontic diseases. A systemic consideration of the patient's overall health along with oral conditions holds the utmost importance in determining the necessity and feasibility of endodontic therapy, as well as selecting appropriate therapeutic approaches. This expert consensus is a collaborative effort by specialists from endodontics and clinical physicians across the nation based on the current clinical evidence, aiming to provide general guidance on clinical procedures, improve patient safety and enhance clinical outcomes of endodontic therapy in patients with compromised overall health.

Functional study on candidate regulators mediating the expression of CYP6B7 induced by fenvalerate in a susceptible strain of Helicoverpa armigera.

Transcription factors play an important role in regulating the expression of detoxification genes (e.g. P450s) that confer insecticide resistance. Our previous study identified a series of candidate transcription factors (CYP6B7-fenvalerate association proteins, CAPs) that may be related to fenvalerate-induced expression of CYP6B7 in a field HDTJ strain of H. armigera. Whether these CAPs can mediate the transcript of CYP6B7 induced by fenvalerate in a susceptible HDS strain of H. armigera remains unknown. Further study showed that the expression levels of multiple CAPs were significantly induced by fenvalerate in HDS strain. Knockdown of CAP19 [fatty acid synthase-like (FAS)], CAP22 [polysaccharide biosynthesis domain-containing protein 1 (PBDC1)], CAP24 [5-formyltetrahydrofolate cycloligase (5-FCL)], CAP30 [peptidoglycan recognition protein LB-like (PGRP)] and CAP33 [NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 11 (NDUFA11)] resulted in significant inhibition of CYP6B7 and some other P450 genes expression; meanwhile, the sensitivity of HDS strain larvae to fenvalerate was significantly increased. In addition, PBDC1, PGRP and NDUFA11, either alone or in combination, could significantly enhance the activity of CYP6B7 promoter in HDS strain, as well as the expression level of CYP6B7 gene in Sf9 cells line. These results suggested that PBDC1, PGRP and NDUFA11 may be involved in the transcript regulation of key detoxifying genes in response to fenvalerate in HDS strain of H. armigera.

Nano-ZnO-modified hydroxyapatite whiskers with enhanced osteoinductivity for bone defect repair.

Hydroxyapatite (HA) whisker (HAw) represents a distinct form of HA characterized by its high aspect ratio, offering significant potential for enhancing the mechanical properties of bone tissue engineering scaffolds. However, the limited osteoinductivity of HAw hampers its widespread application. In this investigation, we observed HAw-punctured osteoblast membranes and infiltrated the cell body, resulting in mechanical damage to cells that adversely impacted osteoblast proliferation and differentiation. To address this challenge, we developed nano-zinc oxide particle-modified HAw (nano-ZnO/HAw). Acting as a reinforcing and toughening agent, nano-ZnO/HAw augmented the compressive strength and ductility of the matrix materials. At the same time, the surface modification with nano-ZnO particles improved osteoblast differentiation by reducing the mechanical damage from HAw to cells and releasing zinc ion, the two aspects collectively promoted the osteoinductivity of HAw. Encouragingly, the osteoinductive potential of 5% nano-ZnO/HAw and 10% nano-ZnO/HAw was validated in relevant rat models, demonstrating the efficacy of this approach in promoting new bone formation in vivo. Our findings underscore the role of nano-ZnO particle surface modification in enhancing the osteoinductivity of HAw from a physical standpoint, offering valuable insights into the development of bone substitutes with favorable osteoinductive properties while simultaneously bolstering matrix material strength and toughness.

Long-term exposure of metamifop affects sex differentiation and reproductive system of zebrafish (Danio rerio).

The extensive use of herbicide metamifop (MET) in rice fields for weeds control will inevitably lead to its entering into water environments and threaten the aquatic organisms. Previous researches have demonstrated that sublethal exposure of MET significantly affected zebrafish development. Yet the long-term toxicological impacts of MET on aquatic life remains unknown. Herein, we investigated the potential effects of MET (5 and 50 µg/L) on zebrafish during an entire life cycle. Since the expression level of male sex differentiation-related gene dmrt1 and sex hormone synthesis-related gene cyp19a1b were significantly changed after 50 µg/L MET exposure for only 7 days, indicators related to sex differentiation and reproductive system were further investigated. Results showed that the transcript of dmrt1 was inhibited, estradiol content increased and testosterone content decreased in zebrafish of both sexes after MET exposure at 45, 60 and 120 dpf. Histopathological sections showed that the proportions of mature germ cells in the gonads of male and female zebrafish (120 dpf) were significantly decreased. Moreover, males had elevated vitellogenin content while females did not after MET exposure; MET induced feminization in zebrafish, with the proportion of females significantly increased by 19.6% while that of males significantly decreased by 13.2% at 120 dpf. These results suggested that MET interfered with the expression levels of gonad development related-genes, disrupted sex hormone balance, and affected sex differentiation and reproductive system of female and male zebrafish, implying it might have potential endocrine disrupting effects after long-term exposure.

Terrirubrum flagellatum gen. nov., sp. nov. of Terrirubraceae fam. nov. and Lichenibacterium dinghuense sp. nov. from forest soil and proposal of Rhodoblastaceae fam. nov.

Two Gram-negative, aerobic, rod-shaped bacterial strains, 7MK25T and 6Y81T, were isolated from forest soil of Dinghushan Biosphere Reserve, Guangdong Province, PR China. Based on the results of 16S rRNA gene sequence analysis, strain 7MK25T showed the highest similarity (93.6 %) to Methyloferula stellata AR4T, followed by Bosea thiooxidans DSM 9653T (93.3 %). Strain 6Y81T had the highest similarity of 97.9 % to Lichenibacterium minor RmlP026T, followed by Lichenibacterium ramalinae RmlP001T (97.2 %). Phylogenomic analysis using the UBCG and PhyloPhlAn methods consistently showed that strain 7MK25T formed a sister clade to Boseaceae, while strain 6Y81T formed an independent clade within the genus Lichenibacterium, both in the order Hyphomicrobiales. The digital DNA-DNA hybridization and average nucleotide identity values between strains 7MK25T, 6Y81T and their close relatives were in the ranges of 19.1-29.9 % and 72.5-85.5 %, respectively. The major fatty acids of 7MK25T were summed feature 8 (C18 : 1 ω7c/C18 : 1 ω6c), C19 : 0 cyclo ω8c, C16 : 0 and C17 : 0 cyclo, while those of 6Y81T were summed feature 8 (C18 : 1 ω7c/C18 : 1 ω6c), C16 : 0 and C16 : 0 3-OH. Strains 7MK25T and 6Y81T took diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and phosphatidylcholine as their dominant polar lipids, and Q-10 as their major respiratory quinone. On the basis of phenotypic and phylogenetic data, strain 7MK25T is proposed to represent a novel species of a novel genus with name Terrirubrum flagellatum gen. nov., sp. nov., within a novel family Terrirubraceae fam. nov., with 7MK25T (=KCTC 62738T=GDMCC 1.1452T) as its type strain. Strain 6Y81T represents a novel species in the genus Lichenibacterium, for which the name Lichenibacterium dinghuense sp. nov. (type strain 6Y81T=KACC 21 727T=GDMCC 1.2176T) is proposed. Rhodoblastaceae fam. nov. with Rhodoblastus as the type genus is also proposed to solve the non-monophylectic problem of the family Roseiarcaceae.

Increased visceral fat area to skeletal muscle mass ratio is positively associated with the risk of metabolic dysfunction-associated steatotic liver disease in a Chinese population.

BACKGROUND: The diagnosis and comprehension of nonalcoholic fatty liver disease (NAFLD), recently redefined as metabolic dysfunction-associated steatotic liver disease (MASLD) are gaining a better understanding. In this study, we examined the association between visceral fat area and skeletal muscle mass ratio (VSR) and the prevalence of MASLD in a Chinese population. METHODS: A cross-sectional study was conducted involving 10,916 individuals who underwent bioelectrical impedance analysis, along with anthropometric and biochemical measurements, from January 2022 to June 2023. According to the VSR distribution, sex-specific quartiles of VSR within the study population were defined. Linear trend tests were performed for the categorized VSR variables. Logistic regression models were performed to estimate the odds ratio and 95% confidence intervals between VSR distribution and MASLD prevalence stratified by sex. RESULTS: The prevalence of MASLD was 37.94% in the overall population (56.34% male), and it gradually increased with higher VSR levels in both genders (P < 0.001). Logistic regression analysis demonstrated a significant association between VSR and MASLD prevalence after adjusting for confounders. The odds ratio (95% confidence interval) for MASLD, comparing the lowest to the highest VSR quartile, was 3.159 (2.671, 3.736) for men and 2.230 (1.764, 2.819) for women (all P < 0.001). Restricted cubic splines also indicated significant non-linear relationships between VSR and MASLD prevalence. CONCLUSIONS: VSR is positively associated with the prevalence of MASLD in this Chinese population, with a notably higher risk for men as VSR increases compared to women.

The molecular characteristics could supplement the staging system of pT2/T3N0M0 esophageal squamous cell carcinoma: a translational study based on a cohort with over 20 years of follow-up.

OBJECTIVE: This study aimed to construct a model based on 23 enrolled molecules to evaluate prognoses of pT2/3N0M0 esophageal squamous cell carcinoma (ESCC) patients with up to 20 years of follow-up. METHODS: The lasso-Cox model was used to identify the candidate molecule. A nomogram was conducted to develop the survival model (molecular score, MS) based on the molecular features. Cox regression and Kaplan-Meier analysis were used in this study. The concordance index (C-index) was measured to compare the predicted ability between different models. The primary endpoint was overall survival (OS). RESULTS: A total of 226 patients and 23 proteins were enrolled in this study. Patients were classified into high-risk (MS-H) and low-risk (MS-L) groups based on the MS score of 227. The survival curves showed that the MS-L cohort had better 5-year and 10-year survival rates than the MS-H group (5-year OS: 51.0% vs. 8.0%; 10-year OS: 45.0% vs. 5.0%, all p < 0.001). Furthermore, multivariable analysis confirmed MS as an independent prognostic factor after eliminating the confounding factors (Hazard ratio 3.220, p < 0.001). The pT classification was confirmed to differentiate ESCC patients' prognosis (Log-rank: p = 0.029). However, the combination of pT and MS could classify survival curves evidently (overall p < 0.001), which showed that the prognostic prediction efficiency was improved significantly by the combination of the pT and MS than by the classical pT classification (C-index: 0.656 vs. 0.539, p < 0.001). CONCLUSIONS: Our study suggested an MS for significant clinical stratification of T2/3N0M0 ESCC patients to screen out subgroups with poor prognoses. Besides, the combination of pT staging and MS could predict survival more accurately for this cohort than the pT staging system alone.

Identification of insect cuticular protein genes LCP17 and SgAbd5 from Helicoverpa armigera and evaluation their roles in fenvalerate resistance.

Insect cuticular protein (ICP) plays an important role in insect growth and development. However, research on the role of ICP in insecticide resistance is very limited. In this study, insect cuticular protein genes LCP17 and SgAbd5 were cloned and characterized in Helicoverpa armigera based on previous transcriptome data. The functions of LCP17 and SgAbd5 genes in fenvalerate resistance were assessed by RNA interference (RNAi), and their response to fenvalerate was further detected. The results showed that LCP17 and SgAbd5 were overexpressed in the fenvalerate-resistant strain comparing with a susceptible strain. The open reading frames of LCP17 and SgAbd5 genes were 423 bp and 369 bp, encoding 141 and 123 amino acids, respectively. LCP17 and SgAbd5 genes were highly expressed in the larval stage, but less expressed in the adult and pupal stages. The expression level of LCP17 and SgAbd5 genes increased significantly after fenvalerate treatment at 24 h. When the cotton bollworms larvae were exposed to fenvalerate at LD50 level, RNAi-mediated silencing of LCP17 and SgAbd5 genes increased the mortality from 50.68% to 68.67% and 63.89%, respectively; the mortality increased to even higher level, which was 73.61%, when these two genes were co-silenced. Moreover, silencing of these two genes caused the cuticle lamellar structure to become loose, which led to increased penetration of fenvalerate into the larvae. The results suggested that LCP17 and SgAbd5 may be involved in the resistance of cotton bollworm to fenvalerate, and LCP17 and SgAbd5 could serve as potential targets for H. armigera control.

Expert consensus on difficulty assessment of endodontic therapy.

Endodontic diseases are a kind of chronic infectious oral disease. Common endodontic treatment concepts are based on the removal of inflamed or necrotic pulp tissue and the replacement by gutta-percha. However, it is very essential for endodontic treatment to debride the root canal system and prevent the root canal system from bacterial reinfection after root canal therapy (RCT). Recent research, encompassing bacterial etiology and advanced imaging techniques, contributes to our understanding of the root canal system's anatomy intricacies and the technique sensitivity of RCT. Success in RCT hinges on factors like patients, infection severity, root canal anatomy, and treatment techniques. Therefore, improving disease management is a key issue to combat endodontic diseases and cure periapical lesions. The clinical difficulty assessment system of RCT is established based on patient conditions, tooth conditions, root canal configuration, and root canal needing retreatment, and emphasizes pre-treatment risk assessment for optimal outcomes. The findings suggest that the presence of risk factors may correlate with the challenge of achieving the high standard required for RCT. These insights contribute not only to improve education but also aid practitioners in treatment planning and referral decision-making within the field of endodontics.

Expert consensus on irrigation and intracanal medication in root canal therapy.

Chemical cleaning and disinfection are crucial steps for eliminating infection in root canal treatment. However, irrigant selection or irrigation procedures are far from clear. The vapor lock effect in the apical region has yet to be solved, impeding irrigation efficacy and resulting in residual infections and compromised treatment outcomes. Additionally, ambiguous clinical indications for root canal medication and non-standardized dressing protocols must be clarified. Inappropriate intracanal medication may present side effects and jeopardize the therapeutic outcomes. Indeed, clinicians have been aware of these concerns for years. Based on the current evidence of studies, this article reviews the properties of various irrigants and intracanal medicaments and elucidates their effectiveness and interactions. The evolution of different kinetic irrigation methods, their effects, limitations, the paradigm shift, current indications, and effective operational procedures regarding intracanal medication are also discussed. This expert consensus aims to establish the clinical operation guidelines for root canal irrigation and a position statement on intracanal medication, thus facilitating a better understanding of infection control, standardizing clinical practice, and ultimately improving the success of endodontic therapy.

Three new species of Agaricus from Baiyun Mountain, Guangzhou, China.

Agaricus is a species-rich genus with more than 600 species around the world. In this work, three new species, Agaricus cacainus, A. baiyunensis, and A. praeclarefibrillosus are described from the specimens collected at Baiyun Mountain, Guangzhou, China, a subtropical area with a monsoon maritime climate, based on phylogenetic analyses and morphological examinations of internal transcribed spacer (ITS1-5.8S-ITS2 = ITS), D1/D2 domains of the large subunit of ribosomal DNA (28S), and a part of translation elongation factor 1-alpha (TEF1). Agaricus cacainus in A. sect. Amoeni is characterized by a parabolic to applanate, slightly depressed pileus covered with chocolate brown, appressed, triangular squamules against white background, a white, furfuraceous stipe, an unchanging context when cut, a fragile and evanescent annulus, usually 4- or 2-spored basidia, and mostly pyriform cheilocystidia. Agaricus baiyunensis in A. sect. Minores has a pileus with a slightly truncate top covered with light brown, downy-wooly fibrillose scales and a light yellowish stipe with membranous annulus. Agaricus praeclarefibrillosus in A. sect. Brunneopicti is characterized by a pileus surface with brownish, triangular, recurved scales and longitudinally splitting lines toward margin, a cottony stipe with white, tiny, recurved fibrils, a single annulus, and variously shaped cheilocystidia, with sparsely ornamented basidiospores. The detailed comparison of their morphological characteristics with closely related species is provided.

Synthesis and Biological Activities of Novel Pyrazole Carboxamides Containing an Aryloxypyridyl Ethylamine Module.

Pyrazole carboxamide is widely utilized in agricultural crop protection. In this research, we synthesized two classes of compounds, namely, pyrazole-5-carboxamide (4a) and pyrazole-4-carboxamide (4b), which are distinguished by the inclusion of the N-1-(6-aryloxypyridin-3-yl) ethylamine skeleton. This design was inspired by the frequent occurrence of diaryl ether modules in pesticide molecules. The bioassay results revealed that some compounds 4a exhibit higher insecticidal activity (IA) than 4b, while some compounds 4b display stronger fungicidal activity compared to 4a. This suggests that pyrazolyl plays a crucial role in determining the selectivity of these compounds toward different biological species. Notably, compound 4a-14 not only retains the potent activity of tolfenpyrad, the exact lead compound of 4a, against Lepidoptera pest Plutella xylostella and Thysanoptera pest Frankliniella occidentalis but also shows excellent IA against pests with piercing-sucking mouthparts, such as Aphis craccivora Koch and Nilaparvata lugens. This research has important implications for the control of pests with piercing-sucking mouthparts and the development of new insecticides and fungicides. The findings highlight the potential of inhibitory complex I as an effective control target for these pests, particularly those that have developed resistance to traditional insecticides. Additionally, it sheds light on the binding mode of 4b-11 and complex II, which serves as a negative reference for the design of SDHI fungicides. The study emphasizes the significance of pyrazolyl in determining selectivity in biological species and identifies avenues for future research in enhancing the biological activity of amino modules. The discovery of (S)-4a-14 not only presents a promising candidate compound for pesticide development but also provides valuable insights into the inhibitory effect of a respiratory chain complex on piercing-sucking insect pests. These findings have important implications in both theory and practice, offering new directions for pest control strategies and pesticide and fungicide development.

Vitamin E alleviates pyraclostrobin-induced toxicity in zebrafish (Danio rerio) and its potential mechanisms.

Strobilurin fungicides (SFs) are commonly used in agriculture worldwide and frequently detected in aquatic environments. High toxicity of SFs to aquatic organisms has caused great concerns. To explore whether vitamin E (VE) can relieve the toxicity caused by pyraclostrobin (PY), zebrafish were exposed to PY with or without VE supplementation. When co-exposure with VE (20 µM), the 96 h-LC50 values of PY to zebrafish embryos, adult, and the 24 h-LC50 value of PY to larvae increased from 43.94, 58.36 and 38.16 µg/L to 64.72, 108.62 and 72.78 µg/L, respectively, indicating that VE significantly decreased the toxicity of PY to zebrafish at different life stages. In addition, VE alleviated the deformity symptoms (pericardial edema and brain damage), reduced speed and movement distance, and decreased heart rate caused by 40 µg/L PY in zebrafish larvae. Co-exposure of PY with VE significantly reduced PY-caused larval oxidative stress and immunotoxicity via increasing the activities of superoxide dismutase, catalase and level of glutathione, as well as reducing the malondialdehyde production and the expression levels of Nrf2, Ucp2, IL-8, IFN and CXCL-C1C. Meanwhile, the expression levels of gria4a and cacng4b genes, which were inhibited by PY, were significantly up-regulated after co-exposure of PY with VE. Moreover, co-exposure with VE significantly reversed the increased mitochondrial DNA copies and reduced ATP content caused by PY in larvae, but had no effect on the expression of cox4i1l and activity of complex III that reduced by PY, suggesting VE can partially improve PY-induced mitochondrial dysfunction. In conclusion, the potential mechanisms of VE alleviating PY-induced toxicity may be ascribed to decreasing the oxidative stress level, restoring the functions of heart and nervous system, and improving the immunity and mitochondrial function in zebrafish.

The ATF4-regulated LncRNA MALAT1 promotes odontoblastic differentiation of human dental pulp stem cells via histone demethylase JMJD3: An in vitro study.

AIM: This study aimed to investigate the upstream regulators and specific mechanisms of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in the odontoblastic differentiation of human dental pulp stem cells (hDPSCs). METHODOLOGY: Human dental pulp stem cells were isolated and cultured, followed by conducting loss- or gain-of-function experiments on ATF4 and loss experiments on MALAT1 to elucidate their respective biological functions in odontoblastic differentiation. Chromatin immunoprecipitation assays and RNA immunoprecipitation were performed to uncover the interaction between ATF4-MALAT1 and MALAT1-JMJD3, respectively. The odontoblastic differentiation was estimated by the mRNA and protein of DSPP and DMP1, as well as alkaline phosphatase staining. RESULTS: Expression of MALAT1 was upregulated in the hDPSCs cultured in an odontoblastic medium, and MALAT1 downregulation suppressed the odontoblastic differentiation of the hDPSCs. Subsequent experiments confirmed that ATF4 promoted odontoblastic differentiation and induced MALAT1 expression by binding to the MALAT1 promoter region. Further experiments revealed that nuclear MALAT1 interacted with JMJD3. MALAT1 knockdown decreased the JMJD3 protein level and demethylase activity, and it enhanced H3K27me3 occupancy of the promoter region of DSPP and DMP1, resulting in the inhibition of DSPP and DMP1 transcription. Importantly, JMJD3 overexpression significantly attenuated the inhibition of odontoblastic differentiation induced by MALAT1 knockdown. CONCLUSIONS: ATF4-regulated MALAT1 plays a positive regulatory role in odontoblastic differentiation of hDPSCs through JMJD3-mediated H3K27me3 modifications of the DSPP and DMP1 promoters.