Effect of formulations and adjuvants on the properties of acetamiprid solution and droplet deposition characteristics sprayed by UAV.

While the pesticide formulations are widely used for pest control, the combined effects of these formulations with adjuvants on droplet behavior, spraying characteristics, and pest control still need to be studied. To clarify their impact on droplet behavior, spraying characteristics, and control efficacy, six formulations of acetamiprid and six adjuvants were examined. A series of laboratory and field experiments were conducted to analyze the physicochemical properties, toxicity against cotton aphids, droplet deposition characteristics, and droplet drift. The results indicated that 5% acetamiprid micro-emulsion (ME) enhanced the physicochemical features and effectiveness in pest control compared to other formulations. The nongjianfei considerably enhanced the efficiency of all acetamiprid formulations when added. The addition of selected adjuvants to pesticide formulations improved the performance of certain physicochemical properties such as viscosity and surface tension and led to higher aphid mortality rates, demonstrating enhanced pest control effectiveness during the present study. In the field experiments, the combination effect of acetamiprid formulations and adjuvants exhibited a higher droplet size, coverage, and density within the cotton canopy. However, 5% acetamiprid ME was found to be most effective followed by nongjianfei. Furthermore, 5% acetamiprid ME with adjuvant reduced the droplet drift and provided better deposition when compared with other formulations. Overall, the combination of specific formulations and adjuvants led to improved physicochemical properties, enhanced droplet deposition characteristics, reduced spray drift, and increased pesticide deposition. These findings highlighted the significance of selecting appropriate pesticide formulations and adjuvants and provided a solid foundation for efficient pesticide spraying through UAVs.

Study on rock fracture mechanism and hydraulic fracturing propagation law of heterogeneous tight sandstone reservoir.

Hydraulic fracturing technology is an effective way to develop tight sandstone reservoirs with low porosity and permeability. The tight sandstone reservoir is heterogeneous and the heterogeneity characteristics has an important influence on fracture propagation. To investigate hydraulic fracture performance in heterogeneous tight reservoir, the X-ray diffraction experiments are carried out, the Weibull distribution method and finite element method are applied to establish the uniaxial compression model and the hydraulic fracture propagation model of heterogeneous tight sandstone. Meanwhile, the sensitivity of different heterogeneity characterization factors and the multi-fracture propagation mechanism during hydraulic fracture propagation is analyzed. The results indicate that the pressure transfer in the heterogeneous reservoir is non-uniform, showing a multi-point initiation fracture mode. For different heterogeneity characterization factors, the heterogeneity characteristics based on elastic modulus are the most sensitive. The multi-fracture propagation of heterogeneous tight sandstone reservoir is different from that of homogeneous reservoir, the fracture propagation morphology is more complex. With the increase of stress difference, the fracture propagation length increases. With the increase of injection rate, the fracture propagation length increases. With the increase of cluster spacing, the propagation length of multiple fractures tends to propagate evenly. This study clarifies the influence of heterogeneity on fracture propagation and provides some guidance for fracturing optimization of tight sandstone reservoirs.

Effects of Different Fermentation Methods on Flavor Quality of Liupao Tea Using GC-Q-TOF-MS and Electronic Nose Analyses.

To further develop Liupao tea products and enhance their flavor, this study investigated the effects of different fermentation methods on the aroma quality of Liupao tea. The aroma quality of Liupao tea was comprehensively analyzed using HS-SPME in combination with GC-Q-TOF-MS, electronic nose, and sensory evaluations. Electronic nose detection showed that the aroma fingerprints of Liupao tea samples with different fermentation methods were different. Sulfides, alcohols, ketones, and methyls were the main aroma categories affecting the aroma of the four groups of Liupao tea samples. GC-Q-TOF-MS analysis revealed significant differences in the composition of aroma components among the four fermentation methods of Liupao tea (p < 0.05). Furthermore, the total amount of aroma compounds was found to be highest in the group subjected to hot fermentation combined with the inoculation of Monascus purpureus (DMl group). Based on the OPLS-DA model, candidate differential aroma components with VIP > 1 were identified, and characteristic aroma compounds were selected based on OAV > 10. The key characteristic aroma compounds shared by the four groups of samples were 1,2,3-Trimethoxybenzene with a stale aroma and nonanal with floral and fruity aromas. The best sensory evaluation results were obtained for the DMl group, and its key characteristic aroma compounds mainly included 1,2,3-Trimethoxybenzene, nonanal, and cedrol. The results of this study can guide the development of Liupao tea products and process optimization.

Study on plant protection unmanned aerial vehicle spraying technology based on the thrips population activity patterns during the cotton flowering period.

Over the years, thrips have transitioned from a minor nuisance to a major problem, significantly impacting the yield and quality of cotton. Unmanned aerial vehicles (UAVs) for plant protection have emerged as an effective alternative to traditional pesticide spraying equipment. UAVs offer advantages such as avoiding crop damage and enhancing pesticide deposition on the plants and have become the primary choice for pesticide application in cotton fields. In this study, a 2-year field experiment found that the thrips population in a cotton field in Xinjiang, China, exhibited gradual growth during the early flowering phase, peaking in late July. The thrips population gradually shifted from the lower canopy to the upper canopy as the cotton flowers opened layer by layer. From 09:00 to 11:00 (GMT+8) and 19:00 to 21:00 (GMT+8), thrips mainly flew outside the flowers, while from 17:00 to 19:00 (GMT+8), they mostly inhabited the inner whorls of flowers. The insecticides 10% cyantraniliprole oil dispersion and 10% spinetoram suspension concentrate, sprayed by UAV, had the best control effect on thrips, with 80.51% and 79.22% control effect after 7 days of spraying, respectively. The optimal spraying time for 10% cyantraniliprole oil dispersion was 19:00 (GMT+8), and the control effect on thrips reached 91.16% at 7 days of spraying. During the cotton flowering period, thrips inhabited flowers in the evening and flew outside during the day. The best control effect on thrips was achieved with UAV-sprayed 10% cyantraniliprole oil dispersion at 19:00 (GMT+8).

Over-commitment positively predicts hair cortisol concentrations only in nurses with high need for recovery.

PURPOSE: In the contemporary workplace, enduring fatigue has become a standard for employees. This investigation assesses whether such working conditions exacerbate the depletion of employees' personal resources. The need for recovery serves as an indicator of the necessity to mitigate post-work fatigue. A high need for recovery signifies that employees must commence a new workday while already fatigued. METHODS: This research recruited two cohorts of nurses, categorized by a high need for recovery and a low need for recovery, to examine the correlation between work effort and hair cortisol concentrations in each group. RESULTS: Hair cortisol concentrations serve as a biological marker of cumulative cortisol secretion over a specific time frame, reflecting overall personal resource expenditure during this interval. Findings revealed a notable positive correlation between intrinsic work effort (over-commitment) and hair cortisol levels exclusively among nurses with a high need for recovery. CONCLUSION: These outcomes imply that active effort amidst fatigue may lead to excessive strain. This insight enriches the classic 'effort-recovery' model by illustrating how an employee' s personal volition can influence the accumulation of fatigue.

Parametric Stability Criteria for Delayed Recurrent Neural Networks via Flexible Delay-Dividing Method.

This article focuses on investigating the stability issue for recurrent neural networks (RNNs) with interval time-varying delays (TVDs) based on a flexible delay-dividing method with parameters, which are related to the delay derivative. First, an interval of delay is separated into parametric subintervals via the linear combination technique. Then, an establishment of Lyapunov-Krasovskii functional (LKF) is connected to the parameters, and a novel linear technology is suggested to dispose of integral terms in the derivatives of the constructed function. Finally, the validity and advantage of the inferred criteria are interpreted by the comparison of representative simulation examples.

Incidence and risk factors of post-operative delirium in glioma patients: A prospective cohort study in general wards.

AIMS: Glioma patients are at high risk for postoperative delirium (POD), yet studies focusing on this population in general neurosurgical ward settings are limited. This paper investigates the incidence of POD and related risk factors in glioma patients hospitalized in general wards. DESIGN: Prospective observational study. METHODS: This prospective study included 133 adult glioma patients hospitalized in the general neurosurgery ward. In addition to collecting routine perioperative general clinical data, patients' psychological status was assessed preoperatively using the Hospital Anxiety and Depression Scale (HADS). POD was assessed within 3 days postoperatively using the Confusion of Consciousness Assessment method, twice daily. The incidence of POD was calculated, and risk factors were identified using logistic regression analysis. RESULTS: The incidence of POD in glioma patients admitted to the general ward was 31.6% (40/133). Multivariate regression revealed advanced age (age > 50 years), frontal lobe tumour, presence of preoperative anxiety or depression, retention of a luminal drain, postoperative pain, indwelling catheter these six factors were independent risk factors for the development of delirium in patients after surgery. CONCLUSION: In general ward settings, supratentorial glioma patients exhibit a high risk of POD. Critical risk factors include preoperative psychological conditions, as well as postoperative pain, drainage and catheterization. Rigorous preoperative evaluations, effective pain management strategies and the integration of humanistic care principles are essential in mitigating the risk of POD for glioma patients. RELEVANCE TO CLINICAL PRACTICE: In general ward settings, this study reveals the high occurrence of POD in glioma patients and identifies preoperative psychological states, age, tumour location and several postoperative factors as significant risk factors for POD, which provides a framework for targeted interventions. By integrating these insights into clinical practice, healthcare teams can better identify glioma patients at risk for POD and implement preventive measures, thereby enhancing recovery and overall care quality for glioma patients in general neurosurgical wards. REPORTING METHOD: This study adheres to the STROBE guidelines, ensuring a transparent and comprehensive reporting of the observational research methodology and results. PATIENT OR PUBLIC CONTRIBUTION: Patients involvement was limited to the provision of data through their participation in the study's assessments and the collection of clinical information. The study did not involve a direct patient or public contribution in the design, conduct, analysis, or interpretation of the data, nor in the preparation of the manuscript.

The Relationship between Droplet Density and the Defoliation Effect of Cotton Harvest Aids.

Spraying harvesting aids is an important step before the mechanical harvesting of cotton. To clarify the direct relationship between the droplet density and the defoliation effect of cotton harvest aid solutions, we evaluated the relationship between the droplet density and the defoliation effect. The determination method and evaluation standard of the number of droplets required per square centimeter to achieve 50% leaves defoliation (DN50) of the harvest aid solution were further explored. The results revealed a linear relationship between the droplet density and the cotton defoliation rate when the spraying volume was 22.5 L/ha and the harvest aid dosage was 1/2 and 2/3 of the recommended dosage. When the harvest aid dosage was 5/6 and 1 times the recommended dosage, the relationship between the droplet density and the defoliation rate of cotton was logarithmic. The DN50 of the low-concentration harvest aid solution (450 L/ha) was significantly higher than that of the high-concentration solution (22.5 L/ha). The addition of spray adjuvant Beidatong significantly reduced the DN50 of cotton harvest aids. The field experiment showed that the droplet density increased with the increase of the spraying volume sprayed by unmanned aerial vehicles. There was a positive correlation between the spraying volume and the defoliation effect after changes in the cotton harvest aid dosage. When the dosage of Mianhai (MH) was 5/6 of the recommended dosage, the defoliation effect at the spraying volumes of 22.5, 27.0, and 30.0 L/ha reached the peak values at 71.54, 78.56, and 83.23%, respectively. This study proposed the concept of DN50 and its determination method. The fitting equations between the droplet density and defoliation effect and between the harvest aid concentration and defoliation effect were established to provide a theoretical basis for the scientific spraying of cotton harvest aid solutions.

A PAM hydrogel surface-coated hydroponic bamboo evaporator with efficient thermal utilization for solar evaporation.

Solar-driven interfacial water purification emerges as a sustainable technology for seawater desalination and wastewater treatment to address the challenge of water scarcity. Currently, the energy losses via radiation and convection to surrounding environment minimize its energy efficiency. Therefore, it is necessary to develop strategies to minimize the heat losses for efficient water purification. Here, a novel evaporator was developed through the in situ gelation of PAM hydrogel on the surface carbonized hydroponic bamboo (PSC) to promote energy efficiency. The inherent porous and layered network structures of bamboo, in synergy with the functional hydration capacity of PAM hydrogel, facilitated adequate water transportation, while reducing evaporation enthalpy. The PAM hydrogel firmly covered on the photothermal layer surface effectively minimized the radiation and convection heat losses, while further harvesting those thermal energy that would otherwise dissipate into the surrounding environment. The reduced thermal conductivity of PSC served as a thermal insulator as well, obstructing heat transfer to bulk water and thus diminishing conduction losses. Consequently, the rational designed PSC could efficiently convert solar energy to purified water, leading to the evaporation of 2.09 kg m-2 h-1, the energy efficiency of 87.6 % under one sun irradiation, and yielding 9.6 kg m-2 fresh water over 11 h outdoor operation. Moreover, the PSC also performs excellent salt rejection, and long-term stability at outdoor experiment. These results demonstrated high and stable solar evaporation performance could be achieved if turning heat losses into a way of extra energy extraction to further enhance the evaporation performance. This strategy appears to be a promising strategy for effective thermal energy management and practical application.

Effects of biotin and coated cobalamin on lactation performance, nutrient digestion and rumen fermentation in Holstein dairy cows.

Biotin (BI) and cobalamin (CA) are essential for rumen propionate production and hepatic gluconeogenesis. The study evaluated the influence of BI or/and coated CA (CCA) on milk performance and nutrient digestion in cows. Sixty Holstein dairy cows were assigned in a 2 × 2 factorial arrangement and randomised block design to four groups. The factors were BI at 0 or 20 mg/day and CCA at 0 or 9 mg CA/day. Dry matter intake increased with BI addition but was unchanged with CCA supply. Addition of BI or CCA increased fat-corrected milk, milk fat and milk protein yields and feed efficiency. Moreover, lactose yield was increased by CCA addition. Dry matter, organic matter, crude protein and acid detergent fibre total-tract digestibility increased for BI or CCA supply. When CCA was supplemented, positive response of neutral detergent fibre digestibility to BI addition was enhanced. Supplementing BI did not affect pH, propionate content and acetate to propionate ratio, but increased total volatile fatty acids (VFA) and acetate contents. Supplementing CCA decreased pH and acetate to propionate ratio, but increased total VFA, acetate and propionate contents. Rumen protease and carboxymethyl-cellulase activities and fungi, bacteria and Butyrivibrio fibrisolvens numbers increased for BI or CCA supply. In addition, protozoa increased for BI addition, and protease activity and Prevotella ruminicola increased for CCA supply. When CCA was supplemented, positive responses of R. albus and Ruminobacter amylophilus numbers to BI addition were enhanced. Blood glucose concentration was unchanged with BI supply, but increased for CCA supply. Blood nonesterified fatty acids and ß-hydroxybutyrate contents reduced with BI or CCA supply. Supplementation with BI or CCA increased blood BI or CA content. The results showed that supplementing BI or/and CCA improved lactation performance and nutrient digestion, and CCA supply did not enhance the lactation performance response to BI supply.

Effects of nanoselenium supplementation on lactation performance, nutrient digestion and mammary gland development in dairy cows.

The objective of this experiment was to evaluate the influence of nanoselenium (NANO-Se) addition on milk production, milk fatty acid synthesis, the development and metabolism regulation of mammary gland in dairy cows. Forty-eight Holstein dairy cows averaging 720 ± 16.8 kg of body weight, 66.9 ± 3.84 d in milk (dry matter intake [DIM]) and 35.2 ± 1.66 kg/d of milk production were divided into four treatments blocked by DIM and milk yields. Treatments were control group, low-Se (LSe), medium-Se (MSe) and high-Se (HSe) with 0, 0.1, 0.2 and 0.3 mg Se, respectively, from NANO-Se per kg dietary dry matter (DM). Production of energy- and fat-corrected milk (FCM) and milk fat quadratically increased (p < 0.05), while milk lactose yields linearly increased (p < 0.05) with increasing NANO-Se addition. The proportion of saturated fatty acids (SFAs) linearly decreased (p < 0.05), while proportions of monounsaturated fatty acids (MUFAs) linearly increased and polyunsaturated fatty acids (PUFAs) quadratically increased. The digestibility of dietary DM, organic matter (OM), crude protein (CP), neutral detergent fiber (NDF) and acid detergent fiber (ADF) quadratically increased (p < 0.05). Ruminal pH quadratically decreased (p < 0.01), while total VFA linearly increased (p < 0.05) with increasing NANO-Se addition. The acetic to propionic ratio decreased (p < 0.05) linearly due to the unaltered acetic molar percentage and a quadratical increase in propionic molar percentage. The activity of CMCase, xylanase, cellobiase and pectinase increased linearly (p < 0.05) following NANO-Se addition. The activity of α-amylase increased linearly (p < 0.01) with an increase in NANO-Se dosage. Blood glucose, total protein, estradiol, prolactin, IGF-1 and Se linearly increased (p < 0.05), while urea nitrogen concentration quadratically decreased (p = 0.04). Moreover, the addition of Se at 0.3 mg/kg from NANO-Se promoted (p < 0.05) mRNA and protein expression of PPARγ, SREBP1, ACACA, FASN, SCD, CCNA2, CCND1, PCNA, Bcl-2 and the ratios of p-ACACA/ACACA and BCL2/BAX4, but decreased (p < 0.05) mRNA and protein expressions of Bax, Caspase-3 and Caspase-9. The results suggest that milk production and milk fat synthesis increased by NANO-Se addition by stimulating rumen fermentation, nutrients digestion, gene and protein expressions concerned with milk fat synthesis and mammary gland development.

Dietary supplementation of sodium butyrate enhances lactation performance by promoting nutrient digestion and mammary gland development in dairy cows.

This experiment was to evaluate the influence of sodium butyrate (SB) addition on milk production, ruminal fermentation, nutrient digestion, and the development and metabolism regulation of the mammary gland in dairy cows. Forty Holstein dairy cows averaging 710 ± 18.5 kg body weight, 72.8 ± 3.66 d in milk (DIM), and 41.4 ± 1.42 kg/d milk production were divided into four treatments blocked by DIM and milk production. Treatments were control group, low SB, medium SB, and high SB with 0, 100, 200 and 300 g/d of SB addition per cow, respectively. The study lasted for 105 d. Production of milk, milk protein and lactose quadratically increased (P < 0.05), while fat-corrected milk, energy-corrected milk and milk fat yields linearly increased (P < 0.05) with increasing SB addition. The digestibility of dietary dry matter, organic matter, and crude protein linearly increased (P < 0.05), whereas the digestibility of ether extract, neutral detergent fibre, and acid detergent fibre quadratically increased (P < 0.05). Ruminal pH quadratically decreased (P = 0.04), while total volatile fatty acids (VFA) quadratically increased (P = 0.03) with increasing SB addition. The acetic acid to propionic acid ratio increased (P = 0.03) linearly due to the unaltered acetic acid molar percentage and a linear decrease in propionic acid molar percentage. Ruminal enzymatic activity of carboxymethyl-cellulase and α-amylase, populations of total bacteria, total anaerobic fungi, total protozoa, Ruminococcus albus, R. flavefaciens, Butyrivibrio fibrisolvens, Fibrobacter succinogenes, and Ruminobacter amylophilus linearly increased (P < 0.05). Blood glucose, urea nitrogen, and non-esterified fatty acids linearly decreased (P < 0.05), while total protein concentration linearly increased (P = 0.04). Moreover, the addition of SB at 200 g/d promoted (P < 0.05) mRNA and protein expression of PPARγ, SREBF1, ACACA, FASN, SCD, CCNA2, CCND1, PCNA, Bcl-2, GPR41, and the ratios of p-Akt/Akt and p-mTOR/mTOR, but decreased (P < 0.05) mRNA and protein expressions of Bax, caspase-3, and caspase-9. The results suggest that milk production and milk fat synthesis increased with SB addition by stimulating rumen fermentation, nutrient digestion, gene and protein expressions concerned with milk fat synthesis and mammary gland development.

Milk Yields and Milk Fat Composition Promoted by Pantothenate and Thiamine via Stimulating Nutrient Digestion and Fatty Acid Synthesis in Dairy Cows.

Considering the synergistic effect of pantothenate and thiamine on the regulation of energy metabolism, this study investigated the influences of coated calcium pantothenate (CCP) and coated thiamine (CT) on milk production and composition, nutrients digestion, and expressions of genes involved in fatty acids synthesis in mammary glands. Forty-four multiparous Chinese Holstein cows (2.8 ± 0.19 of parity, 772 ± 12.3 kg of body weight [BW], 65.8 ± 8.6 days in milk [DIM] and 35.3 ± 1.9 kg/d of milk production, mean ± SD) were blocked by parity, BW, DIM, and milk production, and they were allocated into one of four treatments in a 2 × 2 factorial block design. Additional CCP (0 mg/kg [CCP-] or 55 mg/kg dry matter [DM] of calcium pantothenate from CCP [CCP+]) and CT (0 g/kg [CT-] or 5.3 mg/kg DM of thiamine from CT [CT+]) were hand-mixed into the top one-third of total mixed ration. Both CCP and CT additives increased milk production, fat content, true protein, and lactose by promoting nutrient digestibility. The CCP or/and CT supplementation induced the elevation of C11:0, C12:0, C13:0, C14:0, C14:1, C15:0, C15:1, C16:00, C16:1, C24:00, C24:1 fatty acids, saturated fatty acid, and C4-16 fatty acid contents in milk fat; but it decreased C17-22 fatty acid content. Ruminal total VFA content was increased, but pH was decreased by both additives. The ruminal fermentation pattern was altered, and a tendency of acetate formation was implied by the increased acetate-to-propionate ratio after both additives' supplementation. The expressions of PPARγ, SREBPF1, ACACA, FASN, SCD, and FABP3 mRNAs were enhanced by CCP or CT addition, but the relative expression of LPL mRNA was upregulated by CT addition only. Additionally, blood glucose, triglyceride, insulin-like growth factor-1, and total antioxidant capacity were promoted by both additives. The combination of CCP and CT more effectively increased the ruminal total VFA concentration, the acetate to propionate ratio, and blood glucose level, and decreased ammoniacal nitrogen concentration than that achieved by CCP or CT alone. The results suggested that CCP and CT supplementation stimulated lactation performance by promoting nutrient digestion and fatty acid synthesis in the mammary glands.

Effects of dietary folic acid supplementation on lactation performance and mammary epithelial cell development of dairy cows and its regulatory mechanism.

The experiment investigated the impacts of FA on the proliferation of bovine mammary gland epithelial cells (BMECs) and to investigate the underlying mechanisms. Supplementation of 10 µM FA elevated the mRNA expression of proliferating cell nuclear antigen (PCNA), cyclin A2 and cyclin D1, and protein expression of PCNA and Cyclin A1. The mRNA and protein expression of B-cell lymphoma-2 (BCL2) and the BCL2 to BCL2 associated X 4 (BAX4) ratio elevated, while that of BAX, Caspase-3 and Caspase-9 reduced by FA. Both Akt and mTOR signaling pathways were activated by FA. Moreover, the stimulation of BMECs proliferation, the alteration of proliferative genes and protein expression, the change of apoptotic genes and protein expression, and the activation of mTOR signaling pathway caused by FA were obstructed by Akt inhibitor. Suppression of mTOR with Rapamycin reversed the FA-modulated promotion of BMECs proliferation and change of proliferous genes and protein expression, with no impact on mRNA or proteins expression related to apoptosis and FA-activated Akt signaling pathway. Supplementation of rumen-protected FA in cow diets evaluated milk yields and serum insulin-like growth factor-1 and estradiol levels. The results implied that the proliferation of BMECs was stimulated by FA through the Akt-mTOR signaling pathway.

PTPN2 targets TAK1 for dephosphorylation to improve cellular senescence and promote adipose tissue browning in T2DM.

Introduction: The energy imbalance when energy intake exceeds expenditure acts as an essential factor in the development of insulin resistance (IR). The activity of brown adipose tissue, which is involved in the dissipation of energy via heat expenditure decreases under type 2 diabetic mellitus (T2DM) state when the number of pathological aging adipocytes increases. Protein tyrosine phosphatase non-receptor type 2 (PTPN2) regulates several biological processes by dephosphorylating several cellular substrates; however, whether PTPN2 regulates cellular senescence in adipocytes and the underlying mechanism has not been reported. Methods: We constructed a model of type 2 diabetic mice with PTPN2 overexpression to explore the role of PTPN2 in T2DM. Results: We revealed that PTPN2 facilitated adipose tissue browning by alleviating pathological senescence, thus improving glucose tolerance and IR in T2DM. Mechanistically, we are the first to report that PTPN2 could bind with transforming growth factor-activated kinase 1 (TAK1) directly for dephosphorylation to inhibit the downstream MAPK/NF-κB pathway in adipocytes and regulate cellular senescence and the browning process subsequently. Discussion: Our study revealed a critical mechanism of adipocytes browning progression and provided a potential target for the treatment of related diseases.

[Determination of complexation ability of rhubarb with copper ions by ultra-high performance liquid chromatography].

Gandou decoction (GDD) is a traditional Chinese medicine prescription that has been widely used to treat copper metabolism disorders in China with remarkable clinical effect and lower toxicity. However, evaluation of the complexation ability of copper ions is challenging, which hinders screening and discovery of coordinate active ingredients in GDD. An analytical method is needed to determinate the complexation ability of chemical constituents with copper ions. In this study, a rapid and accurate method based on ultra-high performance liquid chromatography (UHPLC) was developed to determine the complexing ability of rhubarb with copper ions. First, the optimal coordination reaction conditions between active ingredients of rhubarb and copper ions were determined. The samples were separated using an Agilent Eclipse Plus C18 column (50 mm×2.1 mm, 1.8 µm) with 5 µL injection volumes. The mobile phase was gradient eluted with methanol and water containing 0.1% (v/v) phosphoric acid at a flow rate of 0.3 mL/min. The detection wavelength was 254 nm and the column temperature was 30 ℃. Under the optimized chromatographic conditions, the rhubarb constituents were effectively separated. Next, peak areas of rhubarb were calculated before and after the coordination reaction between copper ions. The complexing ability of active ingredients in rhubarb with copper ions was evaluated by calculating the rate of changes of their chromatographic peak areas. Finally, ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was used to identify the coordination active ingredients in rhubarb extract. Focusing on the coordination reaction conditions between active ingredients of rhubarb and copper ions revealed that the active ingredients of rhubarb and copper ions reached equilibrium by coordination reaction at pH 9 for 12 h. Methodological evaluation revealed the good stability and repeatability of the method. Under these conditions, 20 major components of rhubarb were identified by UPLC-Q-TOF-MS. According to the coordination rate of each component and copper ions, eight components with strong coordination were screened out (gallic acid 3-O-ß-D-(6'-O-galloyl)-glucopyranoside, aloe emodin-8-O-ß-D-glucoside, sennoside B, l-O-galloyl-2-O-cinnamoyl-glucoside, chysophanol-8-O-ß-D-(6″-O-acetyl)-glucoside, aloe-emodin, rhein and emodin). The respective complexation rates of the components were 62.50%, 29.94%, 70.58%, 32.77%, 34.61%, 26.07%, 28.73% and 31.78%. Compared with other reported methods, the presently developed method can be used to screen the active ingredients of traditional Chinese medicines that have complexing ability with copper ions, especially in complex mixture systems. This study describes an effective detection technology for evaluating and screening the complexing ability of other traditional Chinese medicines with metal ions.

METTL3 (Methyltransferase Like 3)-Dependent N6-Methyladenosine Modification on Braf mRNA Promotes Macrophage Inflammatory Response and Atherosclerosis in Mice.

BACKGROUND: Atherosclerosis is a chronic inflammatory disease, in which macrophages determine the progression of atherosclerotic plaques. However, no studies have investigated how METTL3 (methyltransferase like 3) in macrophages affects atherosclerotic plaque formation in vivo. Additionally, whether Braf mRNA is modified by METTL3-dependent N6-methyladenosine (m6A) methylation remains unknown. METHODS: We analyzed single-cell sequencing data of atherosclerotic plaques in mice fed with a high fat diet for different periods. Mettl3fl/fl Lyz2cre Apoe-/- mice and littermate control Mettl3fl/fl Apoe-/- mice were generated and fed high fat diet for 14 weeks. In vitro, we stimulated peritoneal macrophages with ox-LDL (oxidized low-density lipoprotein) and tested the mRNA and protein expression levels of inflammatory factors and molecules regulating ERK (extracellular signal-regulated kinase) phosphorylation. To find METTL3 targets in macrophages, we performed m6A-methylated RNA immunoprecipitation sequencing and m6A-methylated RNA immunoprecipitation-qPCR. Further, point mutation experiments were used to explore m6A-methylated adenine. Using RNA immunoprecipitation assay, we explored m6A methylation-writing protein bound to Braf mRNA. RESULTS: In vivo, METTL3 expression in macrophages increased with the progression of atherosclerosis. Myeloid cell-specific METTL3 deletion negatively regulated atherosclerosis progression and the inflammatory response. In vitro, METTL3 knockdown or knockout in macrophages attenuated ox-LDL-mediated ERK phosphorylation rather than JNK (c-Jun N-terminal kinase) and p38 phosphorylation and reduced the level of inflammatory factors by affecting BRAF protein expression. The negative regulation of inflammation response caused by METTL3 knockout was rescued by overexpression of BRAF. In mechanism, METTL3 targeted adenine (39725126 in chromosome 6) on the Braf mRNA. Then, YTHDF1 could bind to m6A-methylated Braf mRNA and promoted its translation. CONCLUSIONS: Myeloid cell-specific Mettl3 deficiency suppressed hyperlipidemia-induced atherosclerotic plaque formation and attenuated atherosclerotic inflammation. We identified Braf mRNA as a novel target of METTL3 in the activation of the ox-LDL-induced ERK pathway and inflammatory response in macrophages. METTL3 may represent a potential target for the treatment of atherosclerosis.

E3 ligase RNF99 negatively regulates TLR-mediated inflammatory immune response via K48-linked ubiquitination of TAB2.

Innate immunity is the first line to defend against pathogenic microorganisms, and Toll-like receptor (TLR)-mediated inflammatory responses are an essential component of innate immunity. However, the regulatory mechanisms of TLRs in innate immunity remain unperfected. We found that the expression of E3 ligase Ring finger protein 99 (RNF99) decreased significantly in peripheral blood monocytes from patients infected with Gram negative bacteria (G-) and macrophages stimulated by TLRs ligands, indicating the role of RNF99. We also demonstrated for the first time, the protective role of RNF99 against LPS-induced septic shock and dextran sodium sulfate (DSS)-induced colitis using RNF99 knockout mice (RNF99-/-) and bone marrow-transplanted mice. In vitro experiments revealed that RNF99 deficiency significantly promoted TLR-mediated inflammatory cytokine expression and activated the NF-κB and MAPK pathways in macrophages. Mechanistically, in both macrophages and HEK293 cell line with TLR4 stably transfection, RNF99 interacted with and degraded TAK1-binding protein (TAB) 2, a regulatory protein of the kinase TAK1, via the lysine (K)48-linked ubiquitin-proteasomal pathway on lysine 611 of TAB2, which further regulated the TLR-mediated inflammatory response. Overall, these findings indicated the physiological significance of RNF99 in macrophages in regulating TLR-mediated inflammatory reactions. It provided new insight into TLRs signal transduction, and offered a novel approach for preventing bacterial infections, endotoxin shock, and other inflammatory ills.

Transcriptomic and metabolomic analyses provide insights into the attenuation of neuroinflammation by nervonic acid in MPTP-stimulated PD model mice.

Nervonic acid is one of the most promising bioactive fatty acids, which is believed to be beneficial for the recovery of human cognitive disorders. However, the detailed neuroprotective effects and mode of action of nervonic acid have not yet been fully elucidated. In this study, we used an MPTP-stimulated mouse Parkinson's disease (PD) model as a target to investigate the neuroprotective effects by behavioral tests and integrative analysis of trancriptomes and metabolomes of PD mouse brain with nervonic acid injections. The KEGG pathway enrichment analysis of transcriptomes showed that the genes involved in neuroinflammation were significantly increased after MPTP induction and have been greatly inhibited by nervonic acid injection, while nervonic acid also greatly improved nerve growth and synaptic plasticity pathways which were significantly downregulated by MPTP. At the same time, the upregulation of oleic acid and arachidonic acid metabolism pathways and the downregulation of amino acid metabolism pathways in metabolomes were particularly highlighted in the nervonic acid protection groups compared with the PD model. Meanwhile, it was found that arachidonic acid, oleic acid and taurine play an important regulatory role in the neuroprotective mechanism of nervonic acid through fatty acid metabolism by integrative analysis. Therefore, our study laid a solid foundation for further studies on the specific role of nervonic acid in the inhibition of PD at the level of metabolic regulation.

Study on Droplet Impact and Spreading and Deposition Behavior of Harvest Aids on Cotton Leaves.

The deposition and spreading of pesticide droplets on the surface of plants is a severe challenge to precise pesticide application, which directly affects the pesticide utilization rate and efficacy. Cotton harvest aids are widely used in machine-picked cotton but the effect of formulation and concentration on the droplet behavior and defoliation effect of cotton defoliants is not clear. To clarify the influence of formulation and concentration on the droplet behavior of cotton defoliants, four formulations (suspension concentrate (SC), water dispersible granule (WG), oil dispersion (OD), and wettable powder (WP)) of cotton defoliants were used to prepare different concentrations of harvest aid solutions, according to the spraying volume. The physicochemical properties, droplet impact, and spreading and deposition behavior were studied. The results indicated that the four kinds of harvest aids have good physicochemical properties and can be wet and spread on cotton leaves. The surface tension of the high-concentration harvest aid solution (the spraying volume was less than 1.2 L/667 m2) was increased, which increased the contact angle and reduced the adhesion tension, adhesion work, and the spreading area. Once the harvest aid solution systems impacted the cotton leaves, it could spread to the maximum in a short time (10 ms). The field experiment showed that the droplet spectrum of harvest aids changed slightly, the coefficient of variation (CV) did not exceed 50%, and the defoliation rate was better when the spraying volume was 1.5 L/667 m2. The correlation and principal component analysis showed that the spraying volume (concentration) and coverage were negatively correlated with the defoliation rate, while the viscosity, diffusion factor, and spreading rate were positively correlated with the defoliation rate. Overall, the use of appropriate spraying volume application in cotton fields can improve the performance of spray, increase the effective deposition and wetting spread of defoliants on cotton leaves, further reduce the dosage of defoliants, and improve pesticide utilization. These results can provide a theoretical basis for the scientific preparation and spraying of cotton harvest aid solutions.