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.

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.

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.

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.

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.

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.

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.

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.

Identification and characterization of both cis- and trans-regulators mediating fenvalerate-induced expression of CYP6B7 in Helicoverpa armigera.

As we known, inducibility is an important feature of P450 genes. Previous studies indicated that CYP6B7 could be induced and involved in fenvalerate detoxification in Helicoverpa armigera. However, the regulatory mechanism of CYP6B7 induced by fenvalerate is still unclear. In this study, CYP6B7 promoter of H. armigera was cloned and the cis-acting element of fenvalerate was identified to be located between -84 and - 55 bp of CYP6B7 promoter. Subsequently, 33 candidate transcription factors (CYP6B7-fenvalerate association proteins, CAPs) that may bind to the cis-acting element were screened and verified by yeast one-hybrid. Among them, the expression levels of several CAPs were significantly induced by fenvalerate. Knockdown of juvenile hormone-binding protein-like (JHBP), RNA polymerase II-associated protein 3 (RPAP3), fatty acid synthase-like (FAS) and peptidoglycan recognition protein LB-like (PGRP) resulted in significant expression inhibition of CYP6B7, and increased sensitivity of H. armigera to fenvalerate. Co-transfection of reporter gene p (-84/-55) with pFast-CAP showed that JHBP, RPAP3 and PGRP could significantly increase the activity of CYP6B7 promoter. These results suggested that trans-acting factors JHBP, RPAP3 and PGRP may bind with cis-acting elements to regulate the expression of CYP6B7 induced by fenvalerate, and play an important role in the detoxification of H. armigera to fenvalerate.

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.

Expert consensus on digital guided therapy for endodontic diseases.

Digital guided therapy (DGT) has been advocated as a contemporary computer-aided technique for treating endodontic diseases in recent decades. The concept of DGT for endodontic diseases is categorized into static guided endodontics (SGE), necessitating a meticulously designed template, and dynamic guided endodontics (DGE), which utilizes an optical triangulation tracking system. Based on cone-beam computed tomography (CBCT) images superimposed with or without oral scan (OS) data, a virtual template is crafted through software and subsequently translated into a 3-dimensional (3D) printing for SGE, while the system guides the drilling path with a real-time navigation in DGE. DGT was reported to resolve a series of challenging endodontic cases, including teeth with pulp obliteration, teeth with anatomical abnormalities, teeth requiring retreatment, posterior teeth needing endodontic microsurgery, and tooth autotransplantation. Case reports and basic researches all demonstrate that DGT stand as a precise, time-saving, and minimally invasive approach in contrast to conventional freehand method. This expert consensus mainly introduces the case selection, general workflow, evaluation, and impact factor of DGT, which could provide an alternative working strategy in endodontic treatment.

Regulation of CncC in insecticide-induced expression of cytochrome P450 CYP9A14 and CYP6AE11 in Helicoverpa armigera.

The expression of many detoxification genes can be regulated by CncC pathway and contributes to insecticide tolerance in insects. Our previous study has demonstrated that the transcripts of CncC and cytochrome P450s (CYP9A14, CYP6AE11) were significantly up-regulated after different insecticides treatment in Helicoverpa armigera. Further study indicated that H2O2, GSH, and MDA contents and antioxidant enzyme activities of H. armigera were enhanced after chlorantraniliprole, cyantraniliprole, indoxacarb, and spinosad exposure. Silencing CncC by RNA interference significantly down-regulated the expression levels of CYP9A14 and CYP6AE11, and increased the susceptibility of dsRNA-injected larvae to λ-cyhalothrin, chlorantraniliprole, and cyantraniliprole. On the contrary, applying CncC agonist curcumin on H. armigera induced the expression of CYP9A14 and CYP6AE11, and enhanced the tolerance of H. armigera to insecticides. Treatment of ROS scavenger N-acetylcysteine on H. armigera reduced the H2O2 content and antioxidant enzyme activities, suppressed the transcripts of CncC, CYP9A14, and CYP6AE11, and decreased the larval tolerance to insecticides. These results demonstrated that the induced-expression of CYP9A14 and CYP6AE11 related with insecticides tolerance in H. armigera was regulated by CncC, which may be activated by ROS generated by insecticides. This study will help to better understand the underlying regulation mechanisms of CncC pathway in H. armigera tolerance to insecticides.

Study on the correlation between lifestyle and negative conversion time in patients diagnosed with coronavirus disease (COVID-19): a retrospective cohort study.

BACKGROUND: As of early December 2022, China eased the coronavirus disease (COVID-19) restriction, affecting over 80% of the country's population and posing a severe threat to public health. Previous studies mostly focused factors on the severity/mortality rate of hospitalized COVID-19 patients, but limited studies explored factors associated with virus-negative conversion, particularly lifestyles. Therefore, the aim of our study was to analyze the correlation between lifestyle factors and the negative conversion time in COVID-19 patients. METHODS: We recruited individuals aged 18 years or older who had a clear time record for both the diagnosis and negative conversion of COVID-19 and completed the electronic questionnaire with no missing data. Dietary data collected from the questionnaire was analyzed using exploratory factor analysis to establish dietary patterns. Age segmentation was performed using restricted cubic spline (RCS) plots. The association between lifestyle factors and the time to negative conversion in different age groups, was assessed using Kaplan-Meier plots and Cox regression analysis. RESULT: Out of 514 participants, all achieved viral negative conversion within a median time of 11 days. Based on nutrient intake, we identified four dietary patterns. The relationship between age and negative conversion rate, as depicted by RCS plots, exhibited an inverted "U" shape. We categorized age into three segments: <35 years, 35-45 years, and ≥ 45 years. For individuals under 35, our study indicated that a higher protein intake was linked to a faster recovery among COVID-19 patients, while medical staff or those receiving prescription treatments exhibited a slower recovery rate (P < 0.05). The 35 ~ 45 age group showed that adequate sleep and physical exercise were associated with a shorter time to negative conversion, whereas southern regions and a higher intake of carbohydrates were related with a longer conversion time (P < 0.05). Among individuals aged ≥ 45 years, the negative conversion time was primarily associated with physical exercise and being a medical staff member(P < 0.05). CONCLUSION: Our research suggests that adequate sleep, physical exercise and a higher protein intake can help alleviate COVID-19 symptoms, while a higher level of carbohydrates intake may hinder recovery from COVID-19.

Trinickia violacea sp. nov. and Trinickia terrae sp. nov., isolated from forest soil.

Two Gram-stain negative, aerobic and rod-shaped bacterial strains, DHOD12T and 7GSK02T, were isolated from forest soil of Dinghushan Biosphere Reserve, Guangdong Province, PR China. Strain DHOD12T grew at 4-42 °C (optimum, 28-33 °C), pH 4.0-8.5 (optimum, pH 5.5-6.5) and in the presence of 0-1.5 % (w/v; optimum, 0-0.5 %)NaCl; while strain 7GSK02T grew at 12-42 °C (optimum, 28-33 °C), pH 4.0-8.5 (optimum, pH 5.0-6.0) and in the presence of 0-0.5 % (w/v; optimum, 0 %) NaCl. Strains DHOD12T and 7GSK02T had the highest 16S rRNA sequence similarities of 98.0 and 98.3 % with the same species Trinickia mobilis DHG64T, respectively, and 98.4 % between themselves. In the 16S rRNA phylogeny, they formed a clade that was sister to a major cluster consisting of all described Trinickia species. Phylogenomic analyses with the UBCG and PhyloPhlAn methods consistently showed that strains DHOD12T and 7GSK02T formed a clade with T. mobilis DHG64T that was a sister of a cluster containing the remainder of the Trinickia species. The DNA G+C contents of strains DHOD12T and 7GSK02T were 63.1 and 64.6 mol%, respectively. Digital DNA-DNA hybridization and average nucleotide identity values of strains DHOD12T, 7GSK02T and their closely related strains were in the ranges of 21.6-31.4 % and 77.1-86.9 %, respectively. These two strains had the same major respiratory quinone, ubiquinone-8, and both had C16 : 0, C17 : 0 cyclo and summed feature 8 (C18 : 1 ω7c/C18 : 1 ω6c) as their major fatty acids. Their major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. Genomic analysis indicated that the two strains could have the potential to degrade aromatic compounds like other Trinickia species. On the basis of phenotypic and phylogenetic results, strains DHOD12T and 7GSK02T represent two novel species of the genus Trinickia, for which the names Trinickia violacea sp. nov. (type strain DHOD12T=LMG 30258T=CGMCC 1.15436T) and Trinickia terrae sp. nov. (type strain 7GSK02T=CGMCC 1.15432T=KCTC 62468T) are proposed.

Effects of different insecticides on transcripts of key genes in CncC pathway and detoxification genes in Helicoverpa armigera.

The CncC pathway regulates the expression of multiple detoxification genes and contributes to the detoxification and antioxidation in insects. Many studies have focused on the impacts of plant allelochemicals on the CncC pathway, whereas studies on the effects of pesticides on key genes involved in this pathway are very limited. In this study, the effects of different types of commonly used insecticides on the transcripts of CncC, Keap1, and Maf and multiple detoxification genes of Helicoverpa armigera were evaluated using real-time quantitative polymerase chain reaction. The results showed that 8 insecticides (bifenthrin, λ-cyhalothrin, chlorantraniliprole, cyantraniliprole, spinosad, indoxacarb, chlorfenapyr, tolfenpyrad, and thiacloprid) significantly induced the expression of CncC and 4 insecticides (cypermethrin, acetamiprid, thiacloprid, and indoxacarb) suppressed the expression of Keap1 both at 24 h and 48 h; meanwhile, the expression levels of Maf were induced by 5 insecticides (fenvalerate, chlorantraniliprole, cyantraniliprole, lufenuron, and tolfenpyrad) at 24 h or 48 h. Multiple detoxification genes, especially cytochrome P450s genes, showed different up-regulation after bifenthrin, λ-cyhalothrin, chlorantraniliprole, cyantraniliprole, indoxacarb, and spinosad treatment for 48 h. Our results suggest that the CncC pathway and detoxification genes can be activated by different insecticides in H. armigera. These results establish a foundation for further studies on the relationship between the CncC pathway and the detoxification genes in H. armigera.

Experts consensus on the procedure of dental operative microscope in endodontics and operative dentistry.

The dental operative microscope has been widely employed in the field of dentistry, particularly in endodontics and operative dentistry, resulting in significant advancements in the effectiveness of root canal therapy, endodontic surgery, and dental restoration. However, the improper use of this microscope continues to be common in clinical settings, primarily due to operators' insufficient understanding and proficiency in both the features and established operating procedures of this equipment. In October 2019, Professor Jingping Liang, Vice Chairman of the Society of Cariology and Endodontology, Chinese Stomatological Association, organized a consensus meeting with Chinese experts in endodontics and operative dentistry. The objective of this meeting was to establish a standard operation procedure for the dental operative microscope. Subsequently, a consensus was reached and officially issued. Over the span of about four years, the content of this consensus has been further developed and improved through practical experience.

Paraburkholderia flagellata sp. nov. and Paraburkholderia adhaesiva sp. nov., two novel species isolated from forest soil in Dinghushan Biosphere Reserve in Guangdong, China.

Two Gram-stain-negative, aerobic, motile and short rod strains, designated 4D117T and ZD32-2T, were isolated from the forest soils. Strains 4D117T and ZD32-2T grew optimally at pH 4.0-6.5, 20-33 °C and pH 4.5-7.0, 33 °C, respectively, and both at 0.5% (w/v) NaCl concentration. Strains 4D117T and ZD32-2T shared the highest 16S rRNA gene sequence similarity with P. acidiphila 7Q-K02T (99.1%) and P. ferrariae NBRC 106233T (98.7%), respectively. The genome size and G + C contents of strains 4D117T and ZD32-2T were 9,002,095 bp, 62.9% and 6,974,420 bp, 61.7%, respectively. The dDDH and ANI values between strains 4D117T, ZD32-2T and closely related Paraburkholderia species were in the ranges of 21.9-51.6% and 82.9-94.4%, and 81.7% and 25.4% between themself, respectively. Functional genomic analysis showed both strains were capable of degrading contaminants, such as benzoate, anthranilic acid and catechol for 4D117T, and benzene and catechol for ZD32-2T, indicating that they may have potentials for soil pollutant treatment. The main polar lipids of strains 4D117T and ZD32-2T were phosphatidylglycerol, phosphatidylethanolamine and diphosphatidylglycerol. Strain 4D117T contained C16:0, C19:0 cyclo ω8c and C18:1 ω7c and/or C18:1 ω6c, while strain ZD32-2T had C16:0 and C17:0 cyclo as their major cellular fatty acids (> 10%). Based on the phenotypic characters and genomic data, strains 4D117T and ZD32-2T represent two novel species of genus Paraburkholderia, for which the names Paraburkholderia flagellata sp. nov. (type strain 4D117T = GDMCC 1.2617T = NBRC 115278T) and Paraburkholderia adhaesiva sp. nov. (type strain ZD32-2T = GDMCC 1.2622T = NBRC 115282T) are proposed.

L-Cysteine hydrochloride inhibits Aspergillus flavus growth and AFB1 synthesis by disrupting cell structure and antioxidant system balance.

Aflatoxin B1 (AFB1) is the most potent known naturally occurring carcinogen and pose an immense threat to food safety and human health. L-Cysteine hydrochloride (L-CH) is a food additive often used as a fruit and vegetable preservative and also to approved bread consistency. In this study, we investigated the effects and mechanisms of L-CH as an antimicrobial on the growth of Aspergillus flavus (A. flavus) and AFB1 biosynthesis. L-CH significantly inhibited A. flavus mycelial growth, affected mycelial morphology and AFB1 synthesis. Furthermore, L-CH induced glutathione (GSH) synthesis which scavenged intracellular reactive oxygen species (ROS). RNA-Seq indicated that L-CH inhibited hyphal branching, and spore and sclerotia formation by controlling cell wall and spore development-related genes. Activation of the GSH metabolic pathway eliminated intracellular ROS, leading to hyphal dwarfing. L-CH treatment downregulated most of the Aflatoxin (AF) cluster genes and aflS, aflR, AFLA_091090 transcription factors. This study provides new insights into the molecular mechanism of L-CH control of A. flavus and AFB1 foundation. We believe that L-CH could be used as a food additive to control AFB1 in foods and also in the environment.

Melatonin attenuates dental pulp stem cells senescence due to vitro expansion via inhibiting MMP3.

OBJECTIVE: We aimed to identify the crucial genes involved in dental pulp stem cell (DPSC) senescence and evaluate the impact of melatonin on DPSC senescence. METHODS: Western blotting, SA-ß-Gal staining and ALP staining were used to evaluate the senescence and differentiation potential of DPSCs. The optimal concentration of melatonin was determined using the CCK-8 assay. Differentially expressed genes (DEGs) involved in DPSC senescence were obtained via bioinformatics analysis, followed by RT-qPCR. Gain- and loss-of-function studies were conducted to explore the role of MMP3 in DPSC in vitro expansion and in response to melatonin. GSEA was employed to analyse MMP3-related pathways in cellular senescence. RESULTS: Treatment with 0.1 µM melatonin attenuated cellular senescence and differentiation potential suppression in DPSCs due to long-term in vitro expansion. MMP3 was a crucial gene in senescence, as confirmed by bioinformatics analysis, RT-qPCR and Western blotting. Furthermore, gain- and loss-of-function studies revealed that MMP3 played a regulatory role in cellular senescence. Rescue assays showed that overexpression of MMP3 reversed the effect of melatonin on senescence. GSEA revealed that the MMP3-dependent anti-senescence effect of melatonin was associated with the IL6-JAK-STAT3, TNF-α-Signalling-VIA-NF-κB, COMPLEMENT, NOTCH Signalling and PI3K-AKT-mTOR pathways. CONCLUSION: Melatonin attenuated DPSC senescence caused by long-term expansion by inhibiting MMP3.