Current Research Status, Opportunities, and Future Challenges of Nine Representative Persistent Herbicides.

Pesticides are extensively utilized in contemporary agriculture to manage pests, enhance crop yields, and sustain productivity. Nevertheless, the persistent herbicide represents a dual-edged weapon. On one hand, their prolonged efficacy enables reduced application frequency during crop growth seasons, resulting in cost savings on labor. However, the presence of these residues within fields poses safety risks to soil quality, sensitive crops in subsequent rotations, agricultural product quality, and the ecological environment. This review presents a comprehensive review on the mechanisms of action, application risks, ecotoxicology, and residue analysis methods of nine representative persistent herbicides (namely, atrazine, imazethapyr, imazapic, mesosulfuron-methyl, halosulfuron-methyl, fomesafen, diflufenican, quinclorac, and pyroxasulfone). The objective is to guide their scientific and rational utilization in agricultural practices while minimizing phytotoxicity risks and effectively monitoring and controlling soil pollution. These can not only provide practical recommendations for mitigating potential plant toxicity and ecological environmental risks but also contribute valuable technical insights for efficient soil pollution monitoring and prevention. Additionally, unaddressed research objectives were also anticipated.

The sublethal concentration of acetamiprid suppresses the population growth of 2 species of wheat aphids, Sitobion miscanthi and Schizaphis graminum (Hemiptera: Aphididae).

Sitobion miscanthi and Schizaphis graminum (Rondani) are the 2 main aphid species that occur simultaneously, causing significant loss to wheat production. Acetamiprid has been used to control a variety of pests, including aphids. In this study, the sublethal effect of acetamiprid on S. miscanthi and S. graminum was evaluated using life-table analyses. The results showed that acetamiprid has a high toxicity to S. miscanthi and S. graminum with a LC50 of 1.90 and 3.58 mg/L at 24 h, respectively. The adult longevity and fecundity of S. miscanthi and S. graminum F0 generation were significantly reduced after being exposed to a sublethal concentration of acetamiprid. Additionally, the sublethal concentration of acetamiprid had negative transgenerational effects on S. miscanthi and S. graminum, which showed a significant decrease in fecundity and population life-table parameters involving age-stage-specific survival rate (sxj), age-specific survival rate (lx), and intrinsic rate of increase (r). Furthermore, the population projections showed that the total population size of S. miscanthi and S. graminum was significantly lower in the aphid group exposed to sublethal concentration of acetamiprid compared to the control group. These results suggest that sublethal concentration of acetamiprid suppresses the population growth of S. miscanthi and S. graminum. This finding is beneficial to the control of wheat aphids, and is important to fully understand the role of acetamiprid in integrated pest management.

SlCarE054 in Spodoptera litura (Lepidoptera: Noctuidae) showed direct metabolic activity to ß-cypermethrin with stereoselectivity.

Carboxylesterases (CarEs) is an important detoxification enzyme system in phase Ⅰ participating in insecticides resistance. In our previous study, SlCarE054, a CarEs gene from lepidoptera class, was screened out to be upregulated in a pyrethroids and organophosphates resistant population. Its overexpression was verified in two field-collected populations of Spodoptera litura (Lepidoptera: Noctuidae) resistant to pyrethroids and organophosphates by qRT-PCR. Spatiotemporal expression results showed that SlCarE054 was highly expressed in the pupae stage and the digestive tissue midgut. To further explore its role in pyrethroids and organophosphates resistance, its metabolism activity to insecticides was determined by UPLC. Its recombinant protein showed significant metabolism activity to cyhalothrin and fenvalerate, but not to phoxim or chlorpyrifos. The metabolic activity of SlCarE054 to ß-cypermethrin showed stereoselectivity, with higher metabolic activity to θ-cypermethrin than the enantiomer α-cypermethrin. The metabolite of ß-cypermethrin was identified as 3-phenoxybenzaldehyde. Further modelling and docking analysis indicated that ß-cypermethrin, cyhalothrin and fenvalerate could bind with the catalytic triad of the 3D structure of SlCarE054. The interaction of ß-cypermethrin with SlCarE054 also showed the lowest binding energy. Our work provides evidence that SlCarE054 play roles in ß-cypermethrin resistance in S. litura.

Genetic associations of childhood and adult BMI on chronic heart failure and ischemic stroke: A Mendelian randomization.

Background: Obesity has been confirmed to be associated with cardiovascular disease, but previous studies have focused on adults, and whether childhood obesity is associated with cardiovascular disease in adulthood needs further research. Objective: This Mendelian randomization (MR) study aimed to investigate the associations of childhood and adult body mass index (BMI) with the risk of chronic heart failure (CHF) and ischemic stroke (IS). Methods: Independent genetic instruments, demonstrating a strong association with exposure at the genome-wide significance level (P < 5 × 10-8), were carefully selected from comprehensive genome-wide association studies conducted within relevant European populations. Summary-level data for CHF and IS were obtained from the EBI database and large consortia of European population. To enhance robustness and generalizability, the analysis was replicated in an East Asian population cohort. Results: According to a MR analysis based on a European population, a higher adult BMI was associated with an increased risk of CHF [(odds ratio (OR) 1.594, 95% confidence interval (CI) 1.483-1.713)] and IS (OR 1.163, 95%CI 1.096-1.233). In addition, higher childhood BMI level was associated with a higher risk of CHF (OR 1.323, 95%CI 1.153-1.524).and the effect was mainly driven by adult BMI. Replication analyses of adult BMI in East Asian populations showed consistent findings that adult BMI was associated with the risk of CHF (OR 2.167, 95%CI 1.786-2.630) and IS (OR 1.259, 95%CI 1.128-1.406). Conclusions: Our study findings provide compelling evidence for the significant influence of adult BMI on the occurrence of CHF and IS. Furthermore, our observations suggest that the positive association between childhood BMI and the risk of CHF in adulthood can largely be attributed to individuals who maintain obesity into later life.

The relationship between processed meat, red meat, and risk of cardiovascular disease and type 2 diabetes: A Mendelian randomization study.

BACKGROUND: Numerous observational studies have indicated a potential association between the consumption of processed and red meat and an increased risk of cardiovascular disease and type 2 diabetes mellitus (T2DM). However, the presence of a causal relationship remains uncertain. Therefore, the purpose of this study is to evaluate the impact of processed meat and red meat (pork, lamb, and beef) on the risk of cardiovascular disease, including coronary artery disease (CAD), hypertension, and stroke, and T2DM, using a Two-Sample Mendelian randomization (MR) analysis. METHODS: MR analysis was conducted using the inverse-variance weighted (IVW), weighted median (WM), and MR Egger methods. To identify heterogeneity and pleiotropy, Cochrane's Q test and MR-Egger test were employed. Additionally, the stability of the MR results was assessed using the leave-one-out method. RESULTS: IVW analyses reveal no causal association between the consumption of processed and red meat and the incidence of CAD, hypertension, stroke, and T2DM (P > 0.05). When considering processed meat intake, heterogeneity is observed in hypertension and stroke outcomes (P < 0.05). For pork intake, heterogeneity is seen in hypertension, stroke, and T2DM (P < 0.05). Lamb intake shows heterogeneity in hypertension and T2DM (P < 0.05). However, other exposures and outcomes examined show no heterogeneity (P > 0.05). No significant pleiotropy is detected for all exposures through an MR-Egger test (P > 0.05). Furthermore, the Leave-one-out test demonstrates the robustness of the results. CONCLUSION: The study discerned no observable impact of red and processed meat consumption on CAD, hypertension, stroke, and T2DM. The findings of this study challenge the prevailing conventional perspective in the field.

This study aimed to investigate whether eating processed and red meat increases the risk of heart disease, high blood pressure, stroke, and type 2 diabetes. Using a genetic analysis method called Mendelian randomization, the research team analyzed genome-wide data to assess meat consumption and its potential health effects.

Impacts of evidence-based nursing combined with enteral nutrition on nutritional status and quality of life in acute cerebral infarction patients: A randomized controlled trial.

OBJECTIVE: To explore the impacts of evidence-based nursing (EBN) combined with enteral nutrition (EN) on nutritional status as well as quality of life in acute cerebral infarction (ACI) patients. METHODS: In this randomized controlled cluster trial, 80 ACI patients admitted to our hospital from January 2021 to December 2022 were selected and randomly separated into study group (SG) and control group (CG), with 40 patients in each group. Patients in CG received routine nursing, and patients in the SG received EBN combined with EN. The neurological function, limb movement ability, nutritional status, anxiety and depression, incidence of complications and quality of life between two groups were compared. RESULTS: After intervention, the NIHSS score in the SG was lower than that in the CG (CG = 5.62 ± 0.56, SG = 3.27 ± 0.33. p < .001). The FMA score in the SG was higher compared with the CG (CG = 52.58 ± 5.32, SG = 68.85 ± 6.87. p < .001). The Hb level, TP level and ALB level in the SG were higher relative to the CG (p < .001). The Self-rating Anxiety Scale (SAS) and Self-rating Depression Scale (SDS) scores in the SG were lower in comparison with the CG (SAS score: CG = 42.32 ± 4.25, SG = 36.28 ± 3.64; SDS score: CG = 48.27 ± 4.85, SG = 40.06 ± 4.05. p < .001). The incidence of complications in SG was lower than that in CG. Finally, we found that SF-36 scores in the SG in all dimensions were higher than those in the CG (Physiological function: CG = 70.23 ± 7.05, SG = 82.71 ± 8.26. Role function: CG = 66.28 ± 6.64, SG73.39 ± 7.36. Physical pain: CG = 70.67 ± 7.06, SG = 82.69 ± 8.29. General health: CG = 58.74 ± 5.86, SG66.62 ± 6.65. Mental health: CG = 53.68 ± 5.37, SG = 62.39 ± 6.31. Energy: CG = 60.75 ± 6.08, SG = 67.87 ± 6.78. Social function: CG = 76.25 ± 7.25, SG = 85.78 ± 8.59. Emotional function: CG = 61.23 ± 6.15, SG = 75.74 ± 7.56. p < .001). CONCLUSION: EBN combined with EN can improve the nutritional status and the quality of life in ACI patients compared with the traditional routine nursing, and is suggested as a valuable strategy for clinical management of ACI.

Rapid Segmentation and Diagnosis of Breast Tumor Ultrasound Images at the Sonographer Level Using Deep Learning.

BACKGROUND: Breast cancer is one of the most common malignant tumors in women. A noninvasive ultrasound examination can identify mammary-gland-related diseases and is well tolerated by dense breast, making it a preferred method for breast cancer screening and of significant clinical value. However, the diagnosis of breast nodules or masses via ultrasound is performed by a doctor in real time, which is time-consuming and subjective. Junior doctors are prone to missed diagnoses, especially in remote areas or grass-roots hospitals, due to limited medical resources and other factors, which bring great risks to a patient's health. Therefore, there is an urgent need to develop fast and accurate ultrasound image analysis algorithms to assist diagnoses. METHODS: We propose a breast ultrasound image-based assisted-diagnosis method based on convolutional neural networks, which can effectively improve the diagnostic speed and the early screening rate of breast cancer. Our method consists of two stages: tumor recognition and tumor classification. (1) Attention-based semantic segmentation is used to identify the location and size of the tumor; (2) the identified nodules are cropped to construct a training dataset. Then, a convolutional neural network for the diagnosis of benign and malignant breast nodules is trained on this dataset. We collected 2057 images from 1131 patients as the training and validation dataset, and 100 images of the patients with accurate pathological criteria were used as the test dataset. RESULTS: The experimental results based on this dataset show that the MIoU of tumor location recognition is 0.89 and the average accuracy of benign and malignant diagnoses is 97%. The diagnosis performance of the developed diagnostic system is basically consistent with that of senior doctors and is superior to that of junior doctors. In addition, we can provide the doctor with a preliminary diagnosis so that it can be diagnosed quickly. CONCLUSION: Our proposed method can effectively improve diagnostic speed and the early screening rate of breast cancer. The system provides a valuable aid for the ultrasonic diagnosis of breast cancer.

Blanking period antiarrhythmic drugs after catheter ablation for atrial fibrillation: a meta-analysis of randomized controlled trials.

Background: Antiarrhythmic drugs (AADs) are frequently prescribed following catheter ablation (CA) for atrial fibrillation (AF). However, to date, there is a lack of large-scale, multicenter controlled studies that have confirmed the efficacy of AADs in reducing the incidence of late recurrence of AF after CA. Furthermore, the optimal duration of short-term use of AADs after CA remains a controversial topic. Methods: PubMed, Embase, Cochrane Library, CNKI, and ClinicalTrials.gov were searched until April 25, 2022. We conducted a meta-analysis of randomized controlled trials (RCTs) to assess the efficacy of blanking period AADs in predicting both early and late recurrence of AF. In addition, Grading of Recommendations Assessment, Development and Evaluation (GRADE) was used to assess the quality of evidence in this meta-analysis. Results: 12 RCTs with 3,625 patients were included in this meta-analysis. Short-term use of AADs after AF ablation reduced the risk of early recurrence of AF compared with the no-AADs group. In the subgroup analysis of AADs use time, it was found that only using AADs for more than 2 months can reduce the early recurrence of AF after CA. However, when compared with the no-AADs group, short-term use of AADs after CA did not reduce the incidence of late recurrence of AF. Conclusions: Short-term use of AADs (more than 2 months) can reduce the early recurrence but not the late recurrence of AF after CA.

Hydrolytic Behavior of Novel Pesticide Broflanilide and Its Dissipative Properties in Different Types of Soils.

All pesticides are toxic by nature and pose short- or long-term safety risks to human or the environment, especially when they were used extensively and absence of safety measures. As a new insecticidal active compound with a novel mechanism of action, there is a serious inadequate of information on the hydrolytic behavior of broflanilide in the aqueous environment, as well as its degradation pattern in agricultural soils. In particular, the effects of temperature and pH of the aqueous environment on its hydrolytic behaviors and the dissipation pattern in different types of agricultural soils were still in a dark box. And the further understanding and insights into this insecticidal active ingredient were being deeply conditioned by these doubts. The hydrolysis behavior of broflanilide and the dissipation pattern in soil were systematically investigated by constructing hydrolysis systems with different temperatures and pH values, and conducting spiking experiments in different types of agricultural soil in the laboratory. The obtained results showed that the longest hydrolysis half-life of 10 mg/L broflanilide at 25 °C was 43.32 h (in pH 4.0 buffer), while it was only 12.84 h in pH 9.0 buffer. In pH 7.0 buffer, the hydrolysis rate of broflanilide exhibited a significant temperature dependence, as shown by the fact that for every 10 °C increase in the system temperature, the corresponding hydrolysis rate will increase about 1.5 times. The dissipation experiments in soils showed that broflanilide was most rapidly dissipated in fluvo-aquic soil (half-life of 1.94 days), followed by lime concretion black soil (half-life of 2.53 days) and cinnamon soil (half-life of 3.11 days), and slower in paddy soil (half-life of 4.03 days). It was indicated that broflanilide was a readily degradable pesticide in both aqueous environment and agricultural soil, and it was significantly affected by the temperature and pH of the system.

Development and validation of a novel risk model for predicting atrial fibrillation recurrence risk among paroxysmal atrial fibrillation patients after the first catheter ablation.

Aims: Several models have been developed to predict the risk of atrial fibrillation (AF) recurrence after radiofrequency catheter ablation (RFCA). However, these models are of poor quality from the start. We, therefore, aimed to develop and validate a predictive model for post-operative recurrence of AF. Materials and methods: In a study including 433 patients undergoing the first circumferential pulmonary vein isolation (CPVI) procedure, independent predictors of AF recurrence were retrospectively identified. Using the Cox regression of designated variables, a risk model was developed in a random sample of 70% of the patients (development cohort) and validated in the remaining (validation cohort) 30%. The accuracy and discriminative power of the predictive models were evaluated in both cohorts. Results: During the established 12 months follow-up, 134 patients (31%) recurred. Six variables were identified in the model including age, coronary artery disease (CAD), heart failure (HF), hypertension, transient ischemic attack (TIA) or cerebrovascular accident (CVA), and left atrial diameter (LAD). The model showed good discriminative power in the development cohort, with an AUC of 0.77 (95% confidence interval [CI], 0.69-0.86). Furthermore, the model shows good agreement between actual and predicted probabilities in the calibration curve. The above results were confirmed in the validation cohort. Meanwhile, decision curve analysis (DCA) for this model also demonstrates the advantages of clinical application. Conclusion: A simple risk model to predict AF recurrence after ablation was developed and validated, showing good discriminative power and calibration.

Acute Hypoxia Stress-Induced Apoptosis in Gill of Japanese Flounder (Paralichthys olivaceus) by Modulating the Epas1/Bad Pathway.

The physiological responses and molecular mechanisms of apoptosis in Japanese flounder under hypoxic stress remain unclear. In the present study, we performed acute hypoxia stress on Japanese flounder (2.39 ± 0.84 mg/L) and detected gills responses in histomorphology and molecular mechanisms. The results showed that the volume of the interlamellar cell mass decreased and the gill lamellae prolonged, indicating the expansion of the respiratory surface area. Additionally, the fluorescence signal of apoptosis increased under hypoxic stress. In addition, the expression of two genes (EPAS1 and Bad) related to apoptosis increased about four-fold and two-fold, respectively, at 6 h of hypoxia. Meanwhile, the result of the dual-luciferase reporter assay showed that EPAS1 is a transcription factor, which could regulate (p < 0.05) the expression of the Bad gene, and we identified the binding site of EPAS1 was the AATGGAAAC sequence located near −766. DNA methylation assay showed that hypoxia affected the methylation status of CpG islands of EPAS1 and Bad genes. All results indicated that hypoxia could activate the EPAS1/Bad signal pathway to induce gill apoptosis of Japanese flounder. Our study provides new light on understanding the molecular mechanism of hypoxia-induced apoptosis in Japanese flounder.

Pharmacological interventions for preventing atrial fibrillation after lung surgery: systematic review and meta-analysis.

BACKGROUND: Postoperative atrial fibrillation/flutter (POAF) is one of the most common cardiac complications after lung surgery. We aimed to assess the safety and efficacy of pharmacological interventions for new-onset POAF prophylaxis in patients with lung cancer after lung surgery. METHODS: PubMed, Embase, Web of Science, Scopus, and the Cochrane Library were searched to identify randomized controlled trials comparing the effects of pharmacological interventions to prevent POAF following lung surgery. RESULTS: A total number of 19 studies with 2,922 participants were included. Pharmacological interventions significantly reduced the incidence of POAF (odds ratio [OR] 0.36, 95% confidence interval [95% CI] 0.26-0.52) while did not increase the incidence of severe pulmonary complications (OR 1.17, 95% CI 0.57-2.41) after lung surgery compared with placebo/usual care. Among different trials, beta-blockers appeared to be the most effective with an OR of 0.13 (95% CI, 0.07-0.27) and a number needed-to-treat (NNT) of 3.63 and was considered safe with no serious adverse events recorded. The risk of POAF decreased from 25.6 to 11.4% (P < 0.001) overall and from 34.2 to 6.7% (P < 0.001) with beta-blockers as monotherapy. Pharmacological interventions did not reduce the 30-day mortality (OR 0.89, 95% CI 0.43-1.84, I2 = 0%), but showed a trend toward reducing major cardiovascular complications including myocardial ischemia/infarction, cardiac arrest, heart failure, and stroke (OR 0.41, 95% CI 0.13-1.29, I2 = 0%). CONCLUSION: Current clinical evidence supports the effectiveness of pharmacological intervention with beta-blockers, amiodarone, magnesium sulfate, or calcium-channel blockers to reduce the incidence of POAF after lung surgery in patients with lung cancer. In the absence of contraindications, prophylaxis with beta-blockers seems to be the most effective of the treatments studied.

Hypoxia Affects HIF-1/LDH-A Signaling Pathway by Methylation Modification and Transcriptional Regulation in Japanese Flounder (Paralichthys olivaceus).

Japanese flounder (Paralichthys olivaceus) responsive mechanisms to hypoxia are still not fully understood. Therefore, we performed an acute hypoxic treatment (dissolved oxygen at 2.07 ± 0.08 mg/L) on Japanese flounder. It was confirmed that the hypoxic stress affected the physiological phenotype through changes in blood physiology (RBC, HGB, WBC), biochemistry (LDH, ALP, ALT, GLU, TC, TG, ALB), and hormone (cortisol) indicators. Hypoxia inducible factor-1 (HIF-1), an essential oxygen homeostasis mediator in organisms consisting of an inducible HIF-1α and a constitutive HIF-1ß, and its target gene LDH-A were deeply studied. Results showed that HIF-1α and LDH-A genes were co-expressed and significantly affected by hypoxic stress. The dual-luciferase reporter assay confirmed that transcription factor HIF-1 transcriptionally regulated the LDH-A gene, and its transcription binding sequence was GGACGTGA located at -2343~-2336. The DNA methylation status of HIF-1α and LDH-A genes were detected to understand the mechanism of environmental stress on genes. It was found that hypoxia affected the HIF-1α gene and LDH-A gene methylation levels. The study uncovered HIF-1/LDH-A signaling pathway responsive mechanisms of Japanese flounder to hypoxia in epigenetic modification and transcriptional regulation. Our study is significant to further the understanding of environmental responsive mechanisms as well as providing a reference for aquaculture.

Research Progress of Myocardial Fibrosis and Atrial Fibrillation.

With the aging population and the increasing incidence of basic illnesses such as hypertension and diabetes (DM), the incidence of atrial fibrillation (AF) has increased significantly. AF is the most common arrhythmia in clinical practice, which can cause heart failure (HF) and ischemic stroke (IS), increasing disability and mortality. Current studies point out that myocardial fibrosis (MF) is one of the most critical substrates for the occurrence and maintenance of AF. Although myocardial biopsy is the gold standard for evaluating MF, it is rarely used in clinical practice because it is an invasive procedure. In addition, serological indicators and imaging methods have also been used to evaluate MF. Nevertheless, the accuracy of serological markers in evaluating MF is controversial. This review focuses on the pathogenesis of MF, serological evaluation, imaging evaluation, and anti-fibrosis treatment to discuss the existing problems and provide new ideas for MF and AF evaluation and treatment.

Predicting Late Recurrence of Atrial Fibrillation After Radiofrequency Ablation in Patients With Atrial Fibrillation: Comparison of C2HEST and HATCH Scores.

Objective: This study was aimed to investigate the risk of recurrence in patients with atrial fibrillation (AF) after radiofrequency ablation and predict risk of recurrence using C2HEST and HATCH scores. Methods: We retrospectively included 322 patients with AF from Second Hospital of Lanzhou University, and 261 patients were included in the analysis finally. They had AF and were admitted for radiofrequency catheter ablation. We compared the ability of C2HEST and HATCH scores to predict recurrence after radiofrequency ablation of AF. The predictive ability of C2HEST and HATCH scores for AF recurrence was estimated by the area under the receiver operating characteristic curve (AUROC). The difference in receiver operating characteristic curve between the two models was compared using the DeLong test. Results: Of the 261 patients included in the analysis, 83 (31.6%) patients suffered a late recurrence of AF after radiofrequency ablation. The risk of postoperative recurrence of AF increased with increasing C2HEST and HATCH scores. The AUROC of C2HEST and HATCH scores in predicting postoperative recurrence of AF was 0.773 (95%CI, 0.713-0.833) and 0.801 (95% CI, 0.740-0.861), respectively. There was no significant difference between the two models in their ability to evaluate patients for postoperative recurrence of AF (DeLong test p-value = 0.36). Among the risk factors in both models, hypertension and heart failure (HF) contributed the most to postoperative recurrence after AF, and higher blood pressure and lower cardiac ejection fraction (EF) were associated with a higher risk of recurrence. Conclusion: Both C2HEST and HATCH scores were significantly associated with the risk of late recurrence after radiofrequency ablation of AF. Besides hypertension and HF contributed the most to postoperative recurrence after AF.

The mechanism of immune related signal pathway Egr2-FasL-Fas in transcription regulation and methylated modification of Paralichthys olivaceus under acute hypoxia stress.

Apoptosis genes Egr2, Fas and FasL are related to immune responses. However, the mechanism of these genes inducing apoptosis in fish are still not very clear. An acute hypoxia treatment (1.73 ± 0.06 mg/L) for 24 h was carried out on Japanese flounder (Paralichthys olivaceus). The increasingly dense apoptotic signals at 3 h, 6 h, 12 h by TUNEL in skeletal muscle indicated that hypoxia could quickly affect muscle growth and development. Furthermore, we concluded that the Egr2-FasL-Fas signal pathway, which was located at the upstream of apoptotic executor protein caspases, was related to the apoptosis by quantitative real-time PCR, protein concentration detection in ELISA and double gene in situ hybridization methods. The mechanism of the pathway was researched in transcription regulation and epigenetic modification by dual-luciferase reporter assay and bisulfite modified method, respectively. Egr2, as a transcription factor, could up-regulate the expression of FasL gene. And its binding site was mainly between -479 to -1 of FasL gene promoter. The 5th CpG dinucleotides (-514) methylation levels in FasL gene were significantly affected by hypoxia, and they were negatively correlated with its expressions. These suggested that the -514 site may be a very important site to regulate the FasL gene expression. Above results, we concluded that hypoxia activated the immune related signal pathway Egr2-FasL-Fas to induced skeletal muscle apoptosis to affect growth and development of Japanese flounder. The study revealed the mechanism of hypoxia induced apoptosis, which could provide a reference for fish immunity and aquaculture management.

DNA methylation in promoter region of immune related genes STAT3 and VEGFA and biochemical parameters change in muscle of Japanese flounder under acute hypoxia.

Acute hypoxic stress can lead to immune response in fish, but the molecular mechanism of muscle immunity in fish under acute hypoxia are still unclear. In this study, we carried out the effect of signal transducer and activator of transcription3(STAT3) and vascular endothelial growth factor A(VEGFA) on muscle immune responses of Japanese flounder (Paralichthys olivaceus) during acute hypoxic stimulation (1.65 ± 0.28mg/L O2; 3h, 6h, 12h, 24h) and reoxygenation (7.30 ± 0.40mg/L O2; R12h, R24h, R48h). In situ hybridization (ISH) showed that STAT3 and VEGFA RNA were co-located in the skeletal muscle of Japanese flounder. Japanese flounder was seriously affected by hypoxia for 3h and 6h. The expression of STAT3 and VEGFA increased significantly. The methylation levels of STAT3 5'UTR region and VEGFA promoter region were significantly lower than those in normoxia group, which was negatively correlated with the expression levels of STAT3 and VEGFA. The enzyme activities (LDH, ALT, AST, ALP) changed significantly. In addition, enzyme-linked immunosorbent assay (ELISA) detected a positive correlation between serum VEGFA concentration and muscle VEGFA mRNA. The current study have shown that Japanese flounder responded to acute hypoxic stress at multiple metabolic levels by changing DNA methylation status and activating transcription factors such as HIF-1α, Nrf2 and STAT3. It is significant for the scientific development of aquaculture through analyzing the effects of hypoxia on biological immunity.

Immune correlates of NF-κB and TNFα promoter DNA methylation in Japanese flounder (Paralichthys olivaceus) muscle and immune parameters change response to vibrio anguillarum infection.

Vibrio anguillarum infection can activate NF-κB/TNFα pathway in the immune organs of fish. Fish muscle is also an important immune organ, but the research on its immune function is few. Our aim was to study regulating mechanism of NF-κB and TNFα gene expressions in the muscle of Japanese flounder (Paralichthys olivaceus) which was under Vibrio anguillarum infection (0, 24, 48, 72 and 96 h). The results showed that the expressions of NF-κB and TNFα increased significantly at 48 h, and there was a significant positive correlation between them. In situ hybridization confirmed the co-existence of NF-κB and TNFα genes in Japanese flounder muscle. Interestingly, the expression of the TNFα gene was regulated by the DNA methylation and its methylation level was negatively correlated with the expression. The lowest methylation level of TNFα occurred at 48 h under Vibrio anguillarum infection (P < 0.05). And more, when the fragment (-2122 âˆ¼ -730) was deleted on TNFα gene promoter, double luciferase activity was the highest, indicating that fragment (-730-0) was the transcription factor binding region. The site (-78 ~ -69) on the fragment (-730-0) binding NF-κB was mutated, and double luciferase activity decreased significantly. The results confirmed that the site (-78 ~ -69) was indeed an important binding site for NF-κB. In addition, the activity of TNFα in the serum of Japanese flounder changed with the prolongation of vibrio anguillarum infection, and the concentration of other immune factors such as ALP, ALT, AST and LDH also changed in the muscle under vibrio anguillarum infection. They all showed a trend of first increasing and then decreasing. Above studies implied that Japanese flounder responded to Vibrio anguillarum infection at the immune level with the change of its methylation status and the activation of transcription factor. By studying the mechanism of immune pathways, understanding the response to immune stress is great significant to the research of fish breeding for disease resistance.

SlGSTE9 participates in the stability of chlorpyrifos resistance in Spodoptera litura.

BACKGROUND: Spodoptera litura is an important agricultural pest and has developed serious resistance to multiple insecticides. The resistance level to several insecticides is reported to be unstable, but the mechanism is less reported. RESULTS: Chlorpyrifos and phoxim resistance level in a field-collected population of S. litura declined continuously from the first to the tenth generation and remained stable at the 11th and 12th generations without insecticide exposure. Synergist experiment showed that diethyl maleate and piperonyl butoxide significantly increased mortality to chlorpyrifos and phoxim in the first and sixth generations, but not in the 12th generation. The expression of 31 identified glutathione S-transferase (GST) genes in the third-instar larvae of S. litura in the first, sixth and 12th generations was determined, and eight genes were seen to decrease significantly in the sixth and 12th generations compared with the first generation. SlGSTe9 was selected for further functional study as it had higher abundance and significantly higher expression in the chlorpyrifos-resistant population than in the susceptible population. The recombinant protein of SlGSTE9 showed metabolism activity to chlorpyrifos in vitro and in Escherichia coli, but not to phoxim. Silencing of SlGSTe9 increased the cumulative mortality to chlorpyrifos significantly. SlGSTE9 also showed antioxidant activity to cumene hydroperoxide. CONCLUSION: Our results suggest that SlGSTe9 is directly involved in chlorpyrifos resistance stability, but not in phoxim. SlGSTE9 may also participate in insecticides resistance by relieving the oxidase stress induced by insecticides. © 2021 Society of Chemical Industry.

Acute hypoxia effects on Keap1/Nrf2 (Mafs)-GST pathway related oxidative metabolism in muscle of Japanese flounder (Paralichthys olivaceus).

Acute hypoxia can aggravate the oxidation metabolism of fish muscle tissue. However, the molecular mechanism of oxidative metabolism in fish muscle under acute hypoxia is not very clear. We carried out effects of a typical oxidative metabolism pathway Keap1/Nrf2 (MafG)-GST on muscle oxidative metabolism of Japanese flounder (Paralichthys olivaceus) during acute hypoxia stimulation (1.65 ± 0.05 mg/L; 1 h, 3 h, 6 h, 12 h, 24 h) and reoxygenation (7.30 ± 0.08 mg/L; R12 h, R24 h, R48 h). The mRNAs of Nrf2 and GST in skeletal muscle were found co-existent, and their expressions were significant increase in 3 h and 6 h. The methylation level of CpG island1 in Nrf2 promoter, whose minimum value appeared at 3 h hypoxia treatment group, was affected by acute hypoxia, and it was negatively correlated with Nrf2 expression. The result suggests that environmental factors may regulate gene expression by epigenetic modification. Dual-luciferase reporter assay showed that GST gene was activated by transcription factor Nrf2, whose transcriptional activation binding region in GST promoter was antioxidant response element located near -980 and -852 sites, and Keap1 and MafG were Nrf2 antagonistic and synergistic factor, respectively. Furthermore, the GST activity changed with hypoxia and reoxygenation treatment in muscle, where other oxidative stress factor (MDA), antioxidant factors (T-AOC, GSH) and antioxidant enzyme activities (GST, SOD, CAT) were also changed. The results of MDA and T-AOC being further different between its hypoxia and normoxia groups (P < 0.05) at 6 h demonstrated that hypoxia stimulation lasting for 6 h would deeply affect Japanese flounder. The study illustrated that Japanese flounder responded to acute hypoxia in multiple metabolic levels by changing methylation status and transcription factor activation. It is significant to understand oxidative metabolic mechanism, analyze organism stress response and promote the scientific development of aquaculture.