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1.
Int J Biol Macromol ; 269(Pt 2): 132147, 2024 May 07.
Artículo en Inglés | MEDLINE | ID: mdl-38719007

RESUMEN

Lignin in biomass plays significant role in substitution of synthetic polymer and reduction of energy expenditure, and the lignin content was usually determined by wet chemical methods. However, the methods' heavy workload, low efficiency, huge consumption of chemicals and use of toxic reagents render them unsuitable for sustainable development and environmental protection. Chinese fir, a prevalent angiosperm tree, holds immense importance for various industries. Since our previous work found that Raman spectroscopy could accurately predict the lignin content in poplar, we propose that the lignin content of Chinese fir can be estimated by similar strategy. The results suggested that the peak at 2895 cm-1 is the optimal choice of internal standard peak and algorithm of XGBoost demonstrates the highest accuracy among all algorithms. Furthermore, transfer learning was successfully introduced to enhance the accuracy and robustness of the model. Ultimately, we report that a machine learning algorithm, combining transfer learning with XGBoost or LightGBM, offers an accurate, high-efficiency and environmental friendly method for predicting the lignin content of Chinese fir using Raman spectra.

2.
Food Chem X ; 22: 101407, 2024 Jun 30.
Artículo en Inglés | MEDLINE | ID: mdl-38711773

RESUMEN

Radish (Raphanus sativus L.) undergoes texture changes in their phy-chemical properties during the long-term dry-salting process. In our study, we found that during the 60-day salting period, the hardness and crispness of radish decreased significantly. In further investigation, we observed that the collaborative action of pectin methylesterase (PME) and polygalacturonase (PG) significantly decreased the total pectin, alkali-soluble pectin (ASP), and chelator-soluble pectin (CSP) content, while increasing the water-soluble pectin (WSP) content. Furthermore, the elevated activities of cellulase and hemicellulase directly led to the notable fragmentation of cellulose and hemicellulose. The above reactions jointly induced the depolymerization and degradation of cell wall polysaccharides, resulting in an enlargement of intercellular spaces and shrinkage of the cell wall, which ultimately led to a reduction in the hardness and crispness of the salted radish. This study provided key insights and guidance for better maintaining textural properties during the dry-salting process of radish.

3.
Plants (Basel) ; 13(6)2024 Mar 12.
Artículo en Inglés | MEDLINE | ID: mdl-38592783

RESUMEN

This study aimed to determine the effects of the nitrogen (N) application period and level on the fate of fertilizer N and the contribution of N absorption and translocation to apple organ N. Two N application periods (labeled by the 15N tracer technique in spring and summer, represented by SP and SU, respectively) and three N levels (N0, MN, and HN) were used to determine the physiological indexes and aboveground, root, and soil 15N content of 4-year-old dwarf ('Red Fuji'/M9T337) and arborized ('Red Fuji'/Malus hupehensis Rehd.) apple trees. The results showed that HN led to shoot overgrowth, which was not conducive to the growth of the apple root system (root length, root tips, root surface area, and root volume) or the improvement of root activity. The contribution of soil N to apple organ N accounted for more than 50%, and the contribution of N application in summer to fruit N was higher than that in spring. Under HN treatment, the proportion of soil N absorbed by trees decreased, while that of fertilizer N increased; however, the highest proportion was still less than 50%, so apple trees were highly dependent on soil N. Under MN treatment, fertilizer N residue was similar to soil N consumption, and soil N fertility maintained a basic balance. Under HN treatment, fertilizer N residue was significantly higher than soil N consumption, indicating that excessive N application increased fertilizer N residue in the soil. Overall, the 15N utilization rate of arborized trees (17.33-22.38%) was higher than that of dwarf trees (12.89-16.91%). A total of 12.89-22.38% of fertilizer 15N was absorbed by trees, 30.37-35.41% of fertilizer 15N remained in the soil, and 44.65-54.46% of fertilizer 15N was lost. The 15N utilization rate and 15N residual rate of summer N application were higher than those of spring N application, and the 15N loss rate was lower than that of spring N application. High microbial biomass N (MBN) may be one of the reasons for the high N utilization rate and the low loss rate of N application in summer.

4.
Front Plant Sci ; 15: 1356257, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38463564

RESUMEN

The gaseous phytohormone ethylene (ETH) plays a key role in plant growth and development, and is a major regulator of phenolic biosynthesis. Light has long been known to influence phytohormone signaling transduction. However, whether light influences the effect of ETH on the phenolic composition of grapes (Vitis vinifera L.) is an open question. Here, the accumulation and composition of anthocyanins and non-anthocyanin phenolics were analyzed in Cabernet Sauvignon grapes under four treatments: light exposure with and without ETH treatment, and box-shading with and without ETH treatment. Both light and ETH promoted ripening, decreased the color index (L*, C*, and h*), and accelerated the color change from green to red and purplish red. Sunlight-exposed grapes had the highest contents of most anthocyanins, flavonols, flavan-3-ols, and hydroxybenzoic acids. In addition, light exposure increased the ratios of 3'5'-substituted/3'-substituted anthocyanins and flavonols, but decreased the ratios of methoxylated/non-methoxylated and acylated/non-acylated anthocyanins and flavan-3-ols. Notably, the effects of ETH were influenced by light exposure. Specifically, ETH treatment promoted anthocyanin and non-anthocyanin biosynthesis in light-exposed grapes, and their increasing multiples were remarkably higher under light-exposed conditions. Furthermore, ETH treatment decreased the ratios of methoxylated/non-methoxylated, 3'5'-substituted/3'-substituted, and acylated/non-acylated anthocyanins and flavan-3-ols in light-exposed grapes, each of which was increased by ETH treatment in shaded grapes. Fifteen differential phenolic components were identified through partial least-squares-discriminant analysis (PLS-DA). Among them, cyanidin-3-O-(cis-6-O-coumaryl)-glucoside, petunidin-3-O-(6-O-acetyl)-glucoside, petunidin-3-O-(trans-6-O-coumaryl)-glucoside, petunidin-3-O-glucoside, myricetin-3-O-galactoside, kaempferol-3-O-galactoside, and kaempferol-3-O-glucoside were the main differential components between ETH treatments under different light conditions. This study contributes to the understanding of the impact of ethylene treatment under dark and light conditions on phenolic synthesis in grape berries.

5.
Technol Cancer Res Treat ; 23: 15330338241234798, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38419562

RESUMEN

Objectives: This study aimed to explore the role of inhibin subunit beta A (INHBA) in the progression of cervical cancer (CCa) and investigate its potential as a therapeutic target. Specifically, the objectives were to assess the expression levels of INHBA in CCa, examine its correlation with patient survival, and elucidate its impact on CCa cell proliferation, cell cycle regulation, migration, invasion, and in vivo tumor growth and metastasis. Methods: To achieve the objectives, we conducted a comprehensive set of experimental methods. INHBA expression in CCa was analyzed, and its association with patient survival was assessed using clinical data. In vitro experiments involved the investigation of INHBA's effects on CCa cell proliferation, cell cycle dynamics, migration, and invasion through the epithelial-mesenchymal transition (EMT) process. Additionally, in vivo experiments were performed to evaluate the influence of INHBA on CCa growth and lung metastasis. Results: The results of this study revealed upregulated expression of INHBA in CCa, with a significant association between high INHBA expression and poor patient survival. Functionally, INHBA was found to promote the proliferation of CCa cells, regulate the cell cycle, and enhance migration and invasion through the EMT process in vitro. Moreover, in vivo experiments demonstrated that INHBA facilitated the growth and lung metastasis of CCa. Conclusion: In conclusion, our findings suggest that INHBA plays a crucial role in the progression of cervical cancer. The upregulation of INHBA is associated with poor patient survival, and its involvement in promoting key aspects of cancer progression makes it a potential therapeutic target for CCa treatment. These results provide valuable insights into the molecular mechanisms underlying CCa and offer a foundation for further exploration of targeted therapeutic interventions.


Asunto(s)
Neoplasias de los Conductos Biliares , Colangiocarcinoma , Neoplasias Pulmonares , Neoplasias del Cuello Uterino , Femenino , Humanos , Neoplasias de los Conductos Biliares/genética , Conductos Biliares Intrahepáticos/patología , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Colangiocarcinoma/genética , Transición Epitelial-Mesenquimal/genética , Regulación Neoplásica de la Expresión Génica , Neoplasias del Cuello Uterino/genética
6.
Food Chem X ; 21: 101232, 2024 Mar 30.
Artículo en Inglés | MEDLINE | ID: mdl-38420507

RESUMEN

Sunlight exposure of grape clusters is frequently reported to influence grape aromas greatly. Among them, the effects of full shading (FS) of clusters on fruit quality and volatile compounds in grape berries has scarcely been investigated. In the present study, the effects of FS from véraison to ripeness on fruit quality and volatile compounds in Cabernet Sauvignon grapes were studied. The results showed that FS treatment reduced fruit size and berry weight, delayed fruit maturity, and decreased the contents of anthocyanins, phenols, and tannins in grape berries. In addition, volatile compounds in grape berries were analyzed, and 55 and 53 volatile compounds were detected in the control (CK) and FS groups, respectively. The results indicated that the concentrations of straight-chain fatty aldehydes, straight-chain fatty alcohols, straight-chain fatty acids, and branched-chain fatty acids, norisoprenoids, and total concentration of volatile compounds were all higher in FS group than in CK group. Specifically, FS treatment had significant promoting effects on the concentrations of ß-damascenone, terpineol, 2-ethyl-1-hexanol, and 2-hexenal, and remarkably decreased the concentrations of geranial, benzeneacetaldehyde, neral, and ethyl acetate. Partial least squares-discriminant analysis (PLS-DA) revealed a clear separation between the control (CK) and FS groups, and showed that 2-hexenal and hexanal were the main characteristic aroma compounds in the FS group. Moreover, an increase in the intensity of fruity, herbaceous, floral, and mushroom aromas was recorded in FS grapes. This study provides new insights into the effects of the exclusion of sunlight exposure on volatile compound accumulation in grape berries.

7.
Small ; 20(7): e2303946, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-37806767

RESUMEN

The key to increasing the rate of oxygen evolution reaction (OER) lies in accelerated four-electron dynamics, while the key to facilitating the development of supercapacitors lies in the design of electrode materials. This paper synthesized manganese-iron Prussian blue (MnFe-PBA@IF) at room temperature, and hexagonal concave structures w ere prepared using a fast-reducing matrix. Interestingly, MnFe-PBA@IF has an amorphous structure favorable to exposing more active surfaces. According to Gibbs free energy calculations on MnFe-PBA, charge depletion of manganese atoms can greatly enhance the adsorption of electron-rich oxygen-containing groups on the surface. Furthermore, the overpotential in 1 m KOH is 280 mV. Also, it can be used as a supercapacitor with a stable operating voltage range of -0.9-0 V and a specific capacity of 1260 F g-1 . This work provides new insights into the synthesis of OER catalysts for Prussian blue ferromanganese at room temperature. Non-gold-bonded adsorption, highly active metal centers and active surfaces are the underlying reasons for the superior performance of supercapacitors. Therefore, Prussian blue with good energy storage performance and high active surface can be used as multifunctional energy storage and conversion electrodes.

8.
Chem Sci ; 14(42): 11727-11736, 2023 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-37920345

RESUMEN

Systematically dissecting the highly dynamic and tightly communicating membrane proteome of living cells is essential for the system-level understanding of fundamental cellular processes and intricate relationship between membrane-bound organelles constructed through membrane traffic. While extensive efforts have been made to enrich membrane proteins, their comprehensive analysis with high selectivity and deep coverage remains a challenge, especially at the living cell state. To address this problem, we developed the cell surface engineering coupling biomembrane fusion method to map the whole membrane proteome from the plasma membrane to various organelle membranes taking advantage of the exquisite interaction between two-dimensional metal-organic layers and phospholipid bilayers on the membrane. This approach, which bypassed conventional biochemical fractionation and ultracentrifugation, facilitated the enrichment of membrane proteins in their native phospholipid bilayer environment, helping to map the membrane proteome with a specificity of 77% and realizing the deep coverage of the HeLa membrane proteome (5087 membrane proteins). Furthermore, membrane N-phosphoproteome was profiled by integrating the N-phosphoproteome analysis strategy, and the dynamic membrane proteome during apoptosis was deciphered in combination with quantitative proteomics. The features of membrane protein N-phosphorylation modifications and many differential proteins during apoptosis associated with mitochondrial dynamics and ER homeostasis were found. The method provided a simple and robust strategy for efficient analysis of membrane proteome, offered a reliable platform for research on membrane-related cell dynamic events and expanded the application of metal-organic layers.

9.
Front Public Health ; 11: 1182631, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37663834

RESUMEN

Background: Confidence and appropriate care approach toward dementia among nursing staff (nurses and care assistants) are crucial manifestations of competency to deal with the complexity of dementia care in long-term care facilities (LTCFs). The purpose of this study was to investigate the confidence in dementia care and care approach among nursing staff in LTCFs in mainland China. Methods: A cross-sectional study design was utilized in LTCFs in Shandong Province, mainland China. A convenient sample included of 317 nursing staff drawn from 15 LTCFs. Survey questions included (a) demographics, (b) dementia knowledge, (c) dementia care confidence, and (d) approach to care for people with dementia. Data were analyzed with descriptive statistics. Factors associated with confidence and care approach for people with dementia were examined using Pearson's correlation and multivariate regression analyses. Results: Dementia care confidence was generally moderate. Factors affecting confidence to care for people with dementia included educational level, months of caring dementia patients, and dementia knowledge. Most nursing staff did not use a person-centered care approach which was significantly associated with their age, dementia-learning experience, and knowledge and confidence toward caring for people with dementia. Conclusion: A positive correlation was identified between confidence to care for people with dementia and nursing staff care approach. Clinical recommendations are provided to further develop education strategies tailored for nursing staff to meet the growing demand for dementia care services.


Asunto(s)
Demencia , Personal de Enfermería , Humanos , Estudios Transversales , Cuidados a Largo Plazo , China , Demencia/terapia
10.
Clin Transl Sci ; 16(11): 2323-2330, 2023 11.
Artículo en Inglés | MEDLINE | ID: mdl-37718502

RESUMEN

In vivo pharmacokinetic simulations and virtual bioequivalence (BE) evaluation of cilostazol have not yet been described for humans. Here, we successfully developed a physiologically based absorption model to simulate plasma concentrations of cilostazol. In addition, virtual population simulations integrating dissolution of 0.3% sodium dodecyl sulfate water media were executed to evaluate the BE of test and reference formulations. Simulation results show that test and reference formulations were bioequivalent among 28 subjects, but not nine subjects, consistent with clinical studies. The model proved to be an important tool to show potential BE for cilostazol. This finding may facilitate understanding of the potential risks during the development of generic products.


Asunto(s)
Modelos Biológicos , Humanos , Equivalencia Terapéutica , Cilostazol , Composición de Medicamentos , Simulación por Computador
11.
Sci Total Environ ; 901: 165962, 2023 Nov 25.
Artículo en Inglés | MEDLINE | ID: mdl-37543329

RESUMEN

Groundwater is widely recognized as a source of lake materials. When it discharges into lakes, phosphorus(P)­iron(Fe) geochemical reactions occur due to environmental changes, affecting P discharge from groundwater. However, redox kinetics of Fe and associated P geochemical processes at the sediment-water interface are not fully understood. Taking Dongting Lake as an example, this study explored Fe and P geochemical processes at the sediment-water interface under groundwater discharge with high Fe and P concentrations. We incubated sediments from Dongting Lake under anoxic-oxic conditions with different initial aqueous P/Fe ratios and pH. Aqueous PO43--P and Fe2+, and solid P and Fe phases in sediments were analyzed, and experimental data were further simulated using numerical reactive models. At the beginning of the experiment, aqueous P and Fe were adsorbed rapidly on sediments. Under anoxic conditions, the Fe reduction rate decreased with decreasing content of poorly crystalline ferric (oxyhydr)oxides, and the addition of aqueous P and Fe at neutral pH enhanced the reduction rate. The increased aqueous P was dominated by desorption caused by sediment Fe reduction and then fixed by gibbsite adsorption and hydroxyapatite precipitation. Under oxic conditions, Fe(II) oxidation under was pH- and (P:Fe)ini-independent, with a sharp rate decline. Furthermore, the final sediment Fe(II) content was higher than the initial content, indicating the formation of a low-oxidizability Fe(II) phase. The P dynamics were dominated by adsorption on the produced Fe-oxides. The numerical models also suggested that heterogeneity in natural sediments promotes hydroxyapatite formation at low pH, but restricts it at high pH. The findings reveal that although aqueous P concentration decreased during groundwater discharge to lakes, PO43--P concentration remained much higher than that in natural lake water, increasing the risk of lake eutrophication. The paper provides references for further understanding of P loading from groundwater discharge into lakes.

12.
Clin Pharmacol Drug Dev ; 12(10): 979-984, 2023 10.
Artículo en Inglés | MEDLINE | ID: mdl-37223878

RESUMEN

Limertinib (ASK120067) is a newly developed third-generation epidermal growth factor receptor tyrosine kinase inhibitor. This phase I, open-label, 2-period crossover study was conducted to evaluate the effect of food on the pharmacokinetics (PK) of limertinib and its active metabolite CCB4580030 in Chinese healthy volunteers (HVs). HVs were randomly assigned (1:1) to receive a single dose of limertinib (160 mg) under the fasted state in period 1 and fed condition in period 2, or vice versa. Twenty-four HVs were enrolled, and 20 HVs completed both study periods. PK were assessed before dosing and ≤72 hours after dosing. PK parameters were analyzed by a noncompartmental method. Limertinib was absorbed faster in the fasted state compared with the fed state. The geometric mean ratios (fed/fast) of maximum concentration, area under the plasma concentration-time curve from time 0 to the last quantifiable concentration, and area under the plasma concentration-time curve from time 0 to infinity for ASK120067 were 145.5%, 145.4%, and 141.9%, respectively. Geometric mean ratios of the PK parameters of CCB4580030 were >125.00% and 90% confidence intervals were outside the preset bioequivalent range. Safety profiles were similar in both prandial states, and limertinib was well tolerated. Food reduced the rate and increased the extent of limertinib absorption following oral administration. Whether limertinib can be administered regardless of prandial state in patients warrants further investigation of efficacy and safety.


Asunto(s)
Antineoplásicos , Pueblos del Este de Asia , Receptores ErbB , Ayuno , Humanos , Estudios Cruzados , Ayuno/fisiología , Voluntarios Sanos , Equivalencia Terapéutica , Receptores ErbB/antagonistas & inhibidores , Antineoplásicos/farmacocinética
13.
Chem Commun (Camb) ; 59(32): 4786-4789, 2023 Apr 18.
Artículo en Inglés | MEDLINE | ID: mdl-37000571

RESUMEN

Here, we established a two-stage isothermal amplification method comprising strand exchange amplification and hyperbranched rolling circle amplification, which only employed one DNA polymerase and was performed in a single closed tube at a constant temperature, providing a promising signal amplification strategy for accurate and rapid pathogen detection in the clinic.


Asunto(s)
Ácidos Nucleicos , Técnicas de Amplificación de Ácido Nucleico/métodos , ADN Polimerasa Dirigida por ADN , Temperatura
14.
Reproduction ; 165(6): 593-603, 2023 06 01.
Artículo en Inglés | MEDLINE | ID: mdl-37000598

RESUMEN

In brief: The appropriate growth and functions of Sertoli cells are crucial to testis development and spermatogenesis in mammals. This study reveals a novel mechanism of follicle-stimulating hormone in immature porcine Sertoli cell proliferation. Abstract: Follicle-stimulating hormone (FSH) is a major Sertoli cell mitogen that binds to the FSH receptor. Sertoli cells are indispensable for testis development and spermatogenesis. However, the regulatory mechanisms of FSH in immature Sertoli cell proliferation have not been determined, particularly in domestic animals. In the present study, we identified the regulatory mechanisms of FSH during immature porcine Sertoli cell proliferation. Transcriptome analysis revealed 114 differentially expressed genes that were induced by FSH treatment, which contains 68 upregulated and 46 downregulated genes. These differentially expressed genes were enriched in multiple pathways, including the Ras signaling pathway. Knockdown of the CC-chemokine receptor 7 (CCR7) gene, which was upregulated by FSH, inhibited cell cycle progression by arresting cells in the G1 phase and reduced the cell proliferation and ERK1/2 phosphorylation. In addition, Kobe0065 inhibited Ras signaling in a similar manner as CCR7 knockdown. Furthermore, FSH abolished the effects of Ras signaling pathway inhibition and CCR7 knockdown. Collectively, FSH promotes immature porcine Sertoli cell proliferation by activating the CCR7/Ras-ERK signaling axis. Our results provide novel insights into the regulatory mechanism of FSH in porcine testis development and spermatogenesis by deciding the fate of immature porcine Sertoli cells.


Asunto(s)
Células de Sertoli , Transducción de Señal , Masculino , Animales , Porcinos , Receptores CCR7/metabolismo , Células de Sertoli/metabolismo , Proliferación Celular , Hormona Folículo Estimulante/farmacología , Hormona Folículo Estimulante/metabolismo , Testículo/metabolismo , Mamíferos/metabolismo
15.
Langmuir ; 39(12): 4530-4536, 2023 Mar 28.
Artículo en Inglés | MEDLINE | ID: mdl-36919933

RESUMEN

Lossless and efficient robotic grasping is becoming increasingly important with the widespread application of intelligent robotics in warehouse transportation, human healthcare, and domestic services. However, current sensors for feedback of grasping behavior are greatly restricted by high manufacturing cost, large volume and mass, complex circuit, and signal crosstalk. To solve these problems, here, we prepare lightweight distance sensor-based reduced graphene oxide (rGO)/MXene-rGO coaxial microfibers with interface buffer to assist lossless grasping of a robotic manipulator. The as-fabricated distance microsensor exhibits a high sensitivity of 91.2 m-1 in the distance range of 50-300 µm, a fast response time of 116 ms, a high resolution of 5 µm, and good stability in 500 cycles. Furthermore, the high-performance and lightweight microsensor is installed on the robotic manipulator to reflect the grasp state by the displacement imposed on the sensor. By establishing the correlation between the microsensing signal and the grasp state, the safe, non-destructive, and effective grasp and release of the target can be achieved. The lightweight and high-powered distance sensor displays great application prospects in intelligent fetching, medical surgery, multi-spindle automatic machines, and cultural relics excavation.

16.
Int J Mol Sci ; 24(2)2023 Jan 10.
Artículo en Inglés | MEDLINE | ID: mdl-36674895

RESUMEN

Calcium/calmodulin-dependent protein kinase (CaMK), a key downstream target protein in the Ca2+ signaling pathway of eukaryotes, plays an important regulatory role in the growth, development and pathogenicity of plant fungi. Three AaCaMKs (AaCaMK1, AaCaMK2 and AaCaMK3) with conserved PKC_like superfamily domains, ATP binding sites and ACT sites have been cloned from Alternaria alternata, However, their regulatory mechanism in A. alternata remains unclear. In this study, the function of the AaCaMKs in the development, infection structure differentiation and pathogenicity of A. alternata was elucidated through targeted gene disruption. The single disruption of AaCaMKs had no impact on the vegetative growth and spore morphology but significantly influenced hyphae growth, sporulation, biomass accumulation and melanin biosynthesis. Further expression analysis revealed that the AaCaMKs were up-regulated during the infection structure differentiation of A. alternata on hydrophobic and pear wax substrates. In vitro and in vivo analysis further revealed that the deletion of a single AaCaMKs gene significantly reduced the A. alternata conidial germination, appressorium formation and infection hyphae formation. In addition, pharmacological analysis confirmed that the CaMK specific inhibitor, KN93, inhibited conidial germination and appressorium formation in A. alternata. Meanwhile, the AaCaMKs genes deficiency significantly reduced the A. alternata pathogenicity. These results demonstrate that AaCaMKs regulate the development, infection structure differentiation and pathogenicity of A. alternata and provide potential targets for new effective fungicides.


Asunto(s)
Fungicidas Industriales , Pyrus , Pyrus/microbiología , Virulencia/genética , Alternaria , Fungicidas Industriales/farmacología , Fungicidas Industriales/metabolismo
17.
Sci Total Environ ; 859(Pt 2): 160432, 2023 Feb 10.
Artículo en Inglés | MEDLINE | ID: mdl-36423848

RESUMEN

Di(2-ethyl-hexyl) phthalate (DEHP), an environmental endocrine disruptor, can destroy the sperm genomic integrity and impairs spermatogenesis. N6-methyladenosine (m6A) is involved in the cellular effects of DEHP. However, the genotoxic effect of DEHP on spermatocytes and the possible role of m6A in this process remain unclear. This study demonstrated that m6A alleviates DEHP genotoxicity in GC-2 cells. In DEHP-treated mice, DNA double-strand breaks (DSBs) were induced in the testis and spermatocytes. To further explore the molecular mechanism of DEHP genotoxicity on spermatocytes, GC-2 cells were exposed to DEHP. DEHP produced distinct genotoxicity and caused DSBs, which led to the inhibition of DNA synthesis and cell cycle arrest. The DNA damage response (DDR) was initiated to repair the DSBs induced by environmentally relevant levels of DEHP (100 µM and 200 µM). During this process, METTL3 upregulated m6A, which facilitated the DDR via stabilizing the DNA damage repair factors (Rad51 and Xrcc5) mRNA to maintain the pro-survival state. Moreover, Mettl3 knockdown partially inhibited DDR. Interestingly, high-dose DEHP (400 µM and 600 µM) directly induced apoptosis rather than the pro-survival state. Altogether: METTL3-mediated m6A participates in maintaining the pro-survival state by upregulating DDR, providing guidance for mitigating the genotoxicity of environment-related level DEHP exposure.


Asunto(s)
Dietilhexil Ftalato , Semen , Masculino , Ratones , Animales , Reparación del ADN , Daño del ADN , Espermatozoides , ADN , Dietilhexil Ftalato/toxicidad
18.
Anal Bioanal Chem ; 415(3): 427-438, 2023 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-36385304

RESUMEN

Nucleic acid amplification tests (NAATs) have become an attractive approach for pathogen detection, and obtaining high-quality nucleic acid extracts from biological samples plays a critical role in ensuring accurate NAATs. In this work, we established an elution-free magnetic bead (MB)-based method by introducing polyethylene-polypropylene glycol (PEPPG) F68 in lysis buffer and using NaOH solution instead of alcohols as the washing buffer for rapid nucleic acid extraction from multiple types of biological samples, including nasopharyngeal swabs, serum, milk, and pork, which bypassed the nucleic acid elution step and allowed the nucleic acid/MB composite to be directly used as the template for amplification reactions. The entire extraction process was able to be completed in approximately 7 min. Even though the nucleic acid/MB composite could not be used for quantitative real-time PCR (qPCR) assays, this elution-free MB-based method significantly improved the sensitivity of the loop-mediated isothermal amplification (LAMP) assay. The sensitivity of the quantitative real-time LAMP (qLAMP) assays combined with this elution-free MB-based method showed an improvement of one to three orders of magnitude compared with qLAMP or qPCR assays combined with the traditional MB-based method. In addition to manual operation, like the traditional MB-based method, this universal, rapid, and facile nucleic acid extraction method also has potential for integration into automated robotic processing, making it particularly suitable for the establishment of an analysis platform for ultrafast and sensitive pathogen detection in various biological samples both in centralized laboratories and at remote sites.


Asunto(s)
Ácidos Nucleicos , Técnicas de Amplificación de Ácido Nucleico/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Fenómenos Magnéticos , Sensibilidad y Especificidad
19.
Cancer Med ; 12(6): 7309-7326, 2023 03.
Artículo en Inglés | MEDLINE | ID: mdl-36479622

RESUMEN

BACKGROUND: SPRY4-IT1 (SPRY4 intronic transcript 1) is a long non-coding RNA (lncRNA) that has been identified as a novel oncogene in various cancers, including glioma. However, its function and underlying mechanism in glioma remain largely unclear. Here, we investigated the role of SPRY4-IT1 in the development of glioma and its underlying mechanism. METHODS: Bioinformatics analysis and RT-qPCR assay were used to examine the expression of SPRY4-IT1 in glioma tissues. The CCK-8, EdU, and Xenograft tumor assays wereperformed to assess the proliferation effect of glioma cells. The tube forming assay and Chick Embryo Chorioallantoic Membrane (CAM) assay were conducted to detect the angiogenesis effect of HUVECs. RNA-sequencing, western blotting, RT-qPCR, ELISA, and IHC assays were employed to verify the regulatory mechanism of the SPRY4-IT1/ miR-101-3p/EZH2/VEGFA axis. RESULTS: Analysis of the TCGA dataset and data from our own cohort demonstrated that SPRY4-IT1 was overexpressed in patients with glioma, and high SPRY4-IT1 expression correlated with poor prognosis. In vitro and in vivo experiments showed that SPRY4-IT1 promoted the proliferation of glioma cells. RNA sequencing and Gene Ontology (GO) enrichment analysis indicated significant enrichment of angiogenesis. HUVEC tube forming assay and CAM assay confirmed that SPRY4-IT1 could induce angiogenesis of glioma cells in vitro and in vivo. Mechanistically, SPRY4-IT1 upregulated EZH2 expression by sponging miR-101-3p to induce VEGFA expression in glioma cells. Moreover, SPRY4-IT1 activated the VEGFR2/AKT/ERK1/2 pathway in HUVECs mediated by glioma cells. Rescue experiments further confirmed that SPRY4-IT1 promoted glioma cell proliferation and angiogenesis via the miR-101-3p/EZH2/VEGFA signaling axis. CONCLUSIONS: Our findings provide compelling evidence showing that SPRY4-IT1 upregulated EZH2 to induce VEGFA by sponging miR-101-3p, thereby achieving cell proliferation and angiogenesis in glioma. Therefore, targeting SPRY4-IT1/miR-101-3p/EZH2/VEGFA axis may improve the outcomes of patients with glioma.


Asunto(s)
Glioma , MicroARNs , ARN Largo no Codificante , Embrión de Pollo , Animales , Humanos , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , Línea Celular Tumoral , Glioma/genética , Proliferación Celular/genética , MicroARNs/genética , Regulación Neoplásica de la Expresión Génica , Movimiento Celular/genética , Factor A de Crecimiento Endotelial Vascular/genética , Factor A de Crecimiento Endotelial Vascular/metabolismo , Proteína Potenciadora del Homólogo Zeste 2/genética , Proteína Potenciadora del Homólogo Zeste 2/metabolismo
20.
Neural Regen Res ; 18(1): 226-232, 2023 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-35799547

RESUMEN

Previous studies have shown that berberine has neuroprotective effects against Alzheimer's disease, including antagonizing tau phosphorylation, and inhibiting acetylcholinesterase activity and neural cell apoptosis. However, its low bioavailability and adverse reactions with conventional administration limit its clinical application. In this study, we prepared berberine nanoliposomes using liposomes characterized by low toxicity, high entrapment efficiency, and biodegradability, and modified them with lactoferrin. Lactoferrin-modified berberine nanoliposomes had uniform particle size and high entrapment efficiency. We used the lactoferrin-modified berberine nanoliposomes to treat a mouse model of Alzheimer's disease established by injection of amyloid-beta 1-42 into the lateral ventricle. Lactoferrin-modified berberine nanoliposomes inhibited acetylcholinesterase activity and apoptosis in the hippocampus, reduced tau over-phosphorylation in the cerebral cortex, and improved mouse behavior. These findings suggest that modification with lactoferrin can enhance the neuroprotective effects of berberine nanoliposomes in Alzheimer's disease.

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