α-Fe2O3/TiO2/Ti3C2Tx Nanocomposites for Enhanced Acetone Gas Sensors.

Metal oxide semi-conductors are widely applied in various fields due to their low cost, easy processing, and good compatibility with microelectronic technology. In this study, ternary α-Fe2O3/TiO2/Ti3C2Tx nanocomposites were prepared via simple hydrothermal and annealing treatments. The composition, morphology, and crystal structure of the samples were studied using XPS, SEM, EDS, XRD, and multiple other testing methods. The gas-sensing measurement results suggest that the response value (34.66) of the F/M-3 sensor is 3.5 times higher than the pure α-Fe2O3 sensor (9.78) around 100 ppm acetone at 220°C, with a rapid response and recovery time (10/7 s). Furthermore, the sensors have an ultra-low detection limit (0.1 ppm acetone), excellent selectivity, and long-term stability. The improved sensitivity of the composites is mainly attributed to their excellent metal conductivity, the unique two-dimensional layered structure of Ti3C2Tx, and the heterojunction formed between the nanocomposite materials. This research paves a new route for the preparation of MXene derivatives and metal oxide nanocomposites.

Preschool children's asthma medication: parental knowledge, attitudes, practices, and adherence.

Introduction: As parents or legal guardians primarily care for children with asthma, understanding their knowledge, attitudes, and practices (KAP) barriers to treatment and medication adherence is of essential importance. This study aimed to analyze the KAP toward asthma medication and adherence among preschool-aged asthmatic children's parents and explore the factors influencing adherence. Methods: This cross-sectional study was conducted between February 2023 and April 2023. Parents of preschool children with asthma were asked to complete the questionnaire containing knowledge, attitude, practice dimensions, and demographic characteristics. The Morisky Medication Adherence Scale (MMAS) was used to investigate adherence. Results: A total of 632 valid questionnaires (154 male and 478 female) were included. Parents showed moderate knowledge (9.49 ± 2.86, 63.27%, possible range: 0-15) and moderate attitudes (26.18 ± 2.51, 74.80%, possible range: 7-35) towards asthma medication, while their practices (27.46 ± 5.26, 91.53%, possible range: 6-30) were proactive; however, medication adherence was low (4.84 ± 1.78, total score: 8). The attitude scores (OR = 1.10, 95% CI: 1.01-1.19, P=0.020), practice scores (OR = 1.16, 95%CI: 1.12-1.21, p < 0.001), and smoking (OR = 1.64, 95%CI: 1.14-2.37, p = 0.008) were associated with medication adherence. Discussion: Preschool-aged asthmatic children's parents showed moderate knowledge, attitudes, and proactive practice toward asthma medication. Continuous training and education programs should be provided for parents to improve asthma medication management in preschool children.

Flexible Multimodal Magnetoresistive Sensors Based on Alginate/Poly(vinyl alcohol) Foam with Stimulus Discriminability for Soft Electronics Using Machine Learning.

Flexible foam-based sensors have attracted substantial interest due to their high specific surface area, light weight, superior deformability, and ease of manufacture. However, it is still a challenge to integrate multimodal stimuli-responsiveness, high sensitivity, reliable stability, and good biocompatibility into a single foam sensor. To achieve this, a magnetoresistive foam sensor was fabricated by an in situ freezing-polymerization strategy based on the interpenetrating networks of sodium alginate, poly(vinyl alcohol) in conjunction with glycerol, and physical reinforcement of core-shell bidisperse magnetic particles. The assembled sensor exhibited preferable magnetic/strain-sensing capability (GF ≈ 0.41 T-1 for magnetic field, 4.305 for tension, -0.735 for bending, and -1.345 for pressing), quick response time, and reliable durability up to 6000 cycles under external stimuli. Importantly, a machine learning algorithm was developed to identify the encryption information, enabling high recognition accuracies of 99.22% and 99.34%. Moreover, they could be employed as health systems to detect human physiological motion and integrated as smart sensor arrays to perceive external pressure/magnetic field distributions. This work provides a simple and ecofriendly strategy to fabricate biocompatible foam-based multimodal sensors with potential applications in next-generation soft electronics.

Computational prediction and experimental validation identify functionally conserved lncRNAs from zebrafish to human.

Functional studies of long noncoding RNAs (lncRNAs) have been hindered by the lack of methods to assess their evolution. Here we present lncRNA Homology Explorer (lncHOME), a computational pipeline that identifies a unique class of long noncoding RNAs (lncRNAs) with conserved genomic locations and patterns of RNA-binding protein (RBP) binding sites (coPARSE-lncRNAs). Remarkably, several hundred human coPARSE-lncRNAs can be evolutionarily traced to zebrafish. Using CRISPR-Cas12a knockout and rescue assays, we found that knocking out many human coPARSE-lncRNAs led to cell proliferation defects, which were subsequently rescued by predicted zebrafish homologs. Knocking down coPARSE-lncRNAs in zebrafish embryos caused severe developmental delays that were rescued by human homologs. Furthermore, we verified that human, mouse and zebrafish coPARSE-lncRNA homologs tend to bind similar RBPs with their conserved functions relying on specific RBP-binding sites. Overall, our study demonstrates a comprehensive approach for studying the functional conservation of lncRNAs and implicates numerous lncRNAs in regulating vertebrate physiology.

Association between dietary calcium, potassium, and magnesium consumption and glaucoma.

BACKGROUND: Calcium (Ca), potassium (K) and magnesium (Mg) may be involved in the occurrence and development of glaucoma by influencing the retinal oxidative stress and regulate blood pressure. However, epidemiological opinions on dietary intake of macroelement related to glaucoma are inconsistent. Herein, this study aims to explore the association between dietary Ca, K, and Mg consumption and glaucoma. METHODS: Data of 7,042 adults aged ≥40 years old who received the glaucoma examinations were extracted from the National Health and Nutrition Examination Survey (NHANES) database from 2005 to 2008 in this cross-sectional study. Univariate and multivariate logistic regression were used to explore the association between dietary Ca, K, and Mg intake and glaucoma with the odd ratios (ORs) and 95% confidence intervals (CIs). We also investigated this relationship in individuals of different age, with/without hypertension and visual field defect (VFD). RESULTS: There were 502 (8.11%) participants had glaucoma. After adjusted for covariables, we found that enough dietary Ca consumption was related to a decreased risk of glaucoma [OR = 0.59, 95%CI: (0.42-0.81)], whether in persons with/without hypertension (all P<0.05). In particular, dietary K intake may be a potential protect factor for glaucoma in non-hypertension populations [OR = 0.47, 95%CI: (0.22-0.99), P = 0.049]. Additionally, hypertension/non-hypertension persons who aged <65 years old or with/without VFD should all pay attention to the enough dietary supplement of Ca, K, and Mg according to their own circumstances. CONCLUSION: Enough dietary Ca, K, and Mg consumption may be potential protect factors of glaucoma that could provide some dietary reference for developing targeted glaucoma prevention and control measures.

Analysis of cytokine levels, cytological findings, and MP-DNA level in bronchoalveolar lavage fluid of children with Mycoplasma pneumoniae pneumonia.

BACKGROUND: The present study was conducted to determine the inflammatory response in the lungs of children with Mycoplasma pneumoniae pneumonia (MPP). METHODS: This study retrospectively analyzed cytokine levels, cytological findings, and M. pneumoniae (MP)-DNA level in the bronchoalveolar lavage fluid (BALF) of 96 children with MPP. The study utilized Spearman's correlation method to evaluate the contribution of BALF and blood parameters in MPP children. RESULTS: (1) A total of 96 MPP children were classified into the Low MP-DNA MPP group (BALF MP-DNA ≤ 105 copies/mL) and the High MP-DNA MPP group (BALF MP-DNA > 105 copies/mL); the Non-fever MPP group (no fever during the entire course of pneumonia) and the Fever MPP group; the Defervescence MPP group (fever had subsided at the time of bronchoscopy) and the Fervescence MPP group; and the Mild MPP group and the Severe MPP group. (2) The High MP-DNA MPP, Fever MPP, Fervescence MPP, and Severe MPP groups had higher levels of interleukin (IL)-6, IL-10, and tumor necrosis factor-α (TNF-α) in their BALF (all p < .05). (3) The proportions of neutrophils and macrophages in the BALF of the High MP-DNA MPP and Fever MPP groups increased and decreased, respectively (all p < .05). (4) In the BALF of MPP children, MP-DNA, IL-6, IL-10, TNF-α, and interferon gamma (IFN-γ) levels positively correlated with neutrophil proportion while negatively correlated with macrophage proportion (all p < .05). (5) The MP-DNA, IL-6, IL-10, TNF-α, and IFN-γ levels in the BALF of MPP children were positively correlated with the levels of C-reactive protein, procalcitonin, lactic dehydrogenase, fibrinogen, and d-dimer, while they were negatively correlated with the albumin level (all p < .05). CONCLUSIONS: In children with MPP, the pulmonary inflammatory immune response was stronger in the High MP-DNA MPP, Fever MPP, Fervescence MPP, and Severe MPP groups. The relationship between pulmonary cytokine levels, MP-DNA load, and serum inflammatory parameters were found to be weak.

A prediction model for the efficacy of continuous positive airway pressure on bronchiolitis.

Objectives: Prediction of the efficacy of continuous positive airway pressure (CPAP) on bronchiolitis is necessary for timely treatment. This study aims to establish a nomogram for efficacy of CPAP on bronchiolitis, and compares accuracy with Pediatric Risk of Mortality III (PRISM III), Brighton Pediatric Early Warning Score (Brighton PEWS) and Pediatric Critical Illness Score (PCIS). Methods: From February 2014 to December 2020, data on children diagnosed with bronchiolitis and treated with CPAP in Chongqing was collected. The nomogram was evaluated by using multivariate logistic regression analysis. We compared the predictive value of model with PRISM III, PEWS and PCIS. Results: A total of 510 children were included. The nomogram prediction model including fever, APTT, white blood cells, serum potassium concentration, lactic acid, immunodeficiency, atelectasis, lung consolidation, congenital airway dysplasia and congenital heart disease was established. The AUC of the nomogram was 0.919 in the training set and 0.947 in the validating set. The model fitted well, as evidenced by the calibration curve and Hosmer-Lemeshow goodness-of-fit test. We discovered that the nomogram significantly performed better than PRISM III, PCIS and PEWS. Conclusions: A nomogram including ten factors for predicting the efficacy of CPAP on bronchiolitis was established. It had higher performance than the PRISM III, PCIS, and PEWS in terms of clinical benefits.

Highly selective NH3 gas sensor based on Co(OH)2/Ti3C2T x nanocomposites operating at room temperature.

Ammonia (NH3) is a common air pollutant and is a biomarker for kidney disease. Therefore, the preparation of ammonia gas sensors with high sensitivity, good selectivity and low operating temperature is of great importance for health protection. Using the in situ electrostatic self-assembly approach, a chemoresistive gas sensor based on Co(OH)2/Ti3C2T x hybrid material was created in this study. The prepared samples were characterized by XRD, XPS, TEM, BET and other testing methods for structure, surface topography and elements. These samples were fabricated into sensors, and the gas sensing properties of the materials were investigated under different test conditions. The results show that the gas response value of the C/M-2 sensor is up to about 14.7%/100 ppm, which is three times the response value of the sensor made of pure MXene to NH3. In addition, the Co(OH)2/Ti3C2T x hybrid sensors exhibit excellent repeatability, high sensitivity under low concentration (less than 5 ppm), fast response/recovery time (29 s/49 s) and long-time stability, which indicates their promising utility in the IoT field.

Effect of Occupational Extremely Low-Frequency Electromagnetic Field Exposure on the Thyroid Gland of Workers: A Prospective Study.

OBJECTIVE: The aim of this study was to investigate the biological effects of occupational extremely low-frequency electromagnetic field (ELF-EMF) exposure on the thyroid gland. METHODS: We conducted a prospective analysis of 85 workers (exposure group) exposed to an ELF-EMF (100 µT, 10-100 Hz) produced by the electromagnetic aircraft launch system and followed up on thyroid function indices, immunological indices, and color Doppler images for 3 years. Additionally, 116 healthy volunteers were randomly selected as controls (control group), the thyroid function of whom was compared to the exposure group. RESULTS: No significant difference was observed in thyroid function between the exposure and control groups. During the follow-up of the exposure group, the serum free triiodothyronine (FT3) level was found to slowly decrease and free thyroxine (FT4) level slowly increase with increasing exposure time. However, no significant difference was found in thyroid-stimulating hormone (TSH) over the three years, and no significant difference was observed in the FT3, FT4 and TSH levels between different exposure subgroups. Furthermore, no significant changes were observed in thyroid autoantibody levels and ultrasound images between subgroups or over time. CONCLUSION: Long-term exposure to ELF-EMF may promote thyroid secretion of T4 and inhibit deiodination of T4 to T3. ELF-EMF has no significant effect on thyroid immune function and morphology.

Laboratory evaluation of the compatibility of Beauveria bassiana with the egg parasitoid Trichogramma dendrolimi (Hymenoptera: Trichogrammatidae) for joint application against the oriental fruit moth Grapholita molesta (Lepidoptera: Tortricidae).

BACKGROUND: The entomopathogenic fungus Beauveria bassiana and the egg parasitoid Trichogramma dendrolimi can both contribute to biological control of the global fruit borer Grapholita molesta. To derive insights for optimizing their combined application in augmentation programs, we assayed fungal pathogenicity to both G. molesta (eggs, larvae and adults) and adult parasitoids, and assessed wasp acquisition and transmission of fungal spores following their emergence from B. bassiana-treated host eggs. RESULTS: Concentrations of 108 conidia mL-1 of B. bassiana and higher caused virtually complete mortality of G. molesta larvae, but less than 30% mortality of eggs, and female moths surviving treatments had reduced fecundity. More than 80% of adult T. dendrolimi survived B. bassiana treatments of 107 conidia mL-1 , and more than 60% survived 109 conidia mL-1 , with surviving females achieving reproductive success equivalent to controls. Parasitoid females preferred healthy eggs over B. bassiana-infected ones in choice tests; wasps emerging from treated eggs carried about 104 conidia each and contributed an additional 11% host mortality in trials combining both agents. CONCLUSION: Our results indicate that combined applications of B. bassiana and T. dendrolimi can have complimentary impacts on G. molesta, the wasps compensating for low fungal pathogenicity to eggs and their progeny potentially aiding in subsequent disease transmission. Although foraging wasps tended to avoid infected eggs, negative interactions between the two agents might be further mitigated by timing B. bassiana applications from late instar larva to early moth stage, and T. dendrolimi releases several days later to coincide with peak oviposition periods. © 2022 Society of Chemical Industry.

Harnessing Friction in Intertwined Structures for High-Capacity Reusable Energy-Absorbing Architected Materials.

Energy-absorbing materials with both high absorption capacity and high reusability are ideal candidates for impact protection. Despite great demands, the current designs either exhibit limited energy-absorption capacities or perform well only for one-time usage. Here a new kind of energy-absorbing architected materials is created with both high absorption capacity and superior reusability, reaching 10 kJ kg-1 per cycle for more than 200 cycles, that is, unprecedentedly 2000 kJ kg-1 per lifetime. The extraordinary performance is achieved by exploiting the rate-dependent frictional dissipation between prestressed stiff cores and a porous soft elastomer, which is reinforced by an intertwined stiff porous frame. The vast interfaces between the cores and elastomer enable high energy dissipation, while the magnitude of the friction force can adapt passively with the loading rate. The intertwined structure prevents stress concentration and ensures no damage and reusability of the constituents after hundreds of loading cycles. The behaviors of the architected materials, such as self-recoverability, force magnitude, and working stroke, are further tailored by tuning their structure and geometry. This design strategy opens an avenue for developing high-performance reusable energy-absorbing materials that enable novel designs of machines or structures.

ZEB1 recruits BRG1 to regulate airway remodelling epithelial-to-mesenchymal transition in asthma.

NEW FINDINGS: What is the central question of this study? The aim was to investigate the function of brahma-related gene-1 (BRG1) in airway remodelling epithelial-to-mesenchymal transition (EMT) of asthma and identify the transcription factor of BRG1 that binds to the protomer of E-cadherin. What is the main finding and its importance? This study highlighted an important molecular mechanism involving chromatin remodelling factor BRG1 that played a crucial role in airway remodelling EMT of asthma and demonstrated that ZEB1 was the key transcription factor recruiting BRG1. This finding might offer new insights into gene-based therapy for asthma. ABSTRACT: Epithelial-to-mesenchymal transition (EMT) of airway remodelling happens in children with asthma. A reduction in the epithelial marker E-cadherin is reported to be one of the initiating factors of EMT. Our previous study showed that chromatin remodelling factor brahma-related gene-1 (BRG1) could regulate the expression of E-cadherin indirectly. However, the transcription factor involved in the recruitment of BRG1 in asthma is unknown. Here, we studied the function of Brg1 in an ovalbumin-induced asthma model [lung-specific conditional Brg1 knockdown (Brg1-/- ) mice] and human bronchial epithelial 16HBE cells stably expressing BRG1 short hairpin RNA. Our results showed that Brg1 was involved in EMT in asthmatic mice by detecting the expression of EMT markers. We also identified that BRG1 participated in the transforming growth factor-ß-induced EMT of 16HBE cells. We observed that zinc finger E-box binding homeobox 1 (ZEB1) and BRG1 co-localized in the EMT of TGF-ß-induced 16HBE cells. Further results revealed that ZEB1 recruited BRG1 and bound to the promoter region (+3563/3715) to regulate E-cadherin expression. Thus, ZEB1 might be the key transcription factor to recruit BRG1 in airway remodelling EMT of asthma and might be a new therapeutic target.

Identification and characterization of long non-coding RNA Carip in modulating spatial learning and memory.

CaMKII has long been known to be a key effector for synaptic plasticity. Recent studies have shown that a variety of modulators interact with the subunits of CaMKII to regulate the long-term potentiation (LTP) of hippocampal neurons. However, whether long non-coding RNAs modulate the activity of CaMKII and affect synaptic plasticity is still elusive. Here, we identify a previously uncharacterized long non-coding RNA Carip that functions as a scaffold, specifically interacts with CaMKIIß, and regulates the phosphorylation of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and N-methyl-d-aspartate (NMDA) receptor subunits in the hippocampus. The absence of Carip causes dysfunction of synaptic transmission and attenuates LTP in hippocampal CA3-CA1 synapses, which further leads to impairment of spatial learning and memory. In summary, our findings demonstrate that Carip modulates long-term synaptic plasticity by changing AMPA receptor and NMDA receptor activities, thereby affecting spatial learning and memory in mice.

Embryotoxic effects of tribromophenol on early post-implantation development of mouse embryos in vitro.

2,4,6-Tribromophenol (TBP, CAS No. 118-79-6), the most widely produced brominated phenol, is frequently detected in environmental components. The detection of TBP in human bodies has earned great concerns about its adverse effects on human beings, especially for early embryonic development. Here, we optimized the mouse embryo in vitro culture (IVC) system for early post-implantation embryos and employed it to determine the embryotoxicity of TBP. With this new research model, we revealed the dose-dependent toxic effects of TBP on mouse embryos from peri-implantation to egg cylinder stages. Furthermore, TBP exposure inhibited the differentiation and survival of epiblast (EPI) cells and extraembryonic endoderm (ExEn) cells, while those of extraembryonic ectoderm (ExEc) cells were not influenced. These results implied that TBP might inhibit embryonic development by influencing the generation of three primary germ layers and fetal membranes (the amnion, chorionic disk, umbilical cord, and yolk sac). In summary, we showed a proof of concept for applying mouse embryo IVC system as a novel research model for studying mammalian embryonic toxicology of environmental pollutants. This study also demonstrated the toxicity of TBP on early embryonic development of mammals.

Autoimmune encephalitis with coexistent LGI1 and GABABR1 antibodies: case report.

BACKGROUND: Autoimmune encephalitis (AE) with multiple auto-antibodies is of great clinical significance because its complex clinical manifestations and atypical imaging increase the difficulty of diagnosis, differential diagnosis and treatment, which may aggravate the disease, increase the recurrence rate and mortality. The coexistence of anti-Leucinie-rich Glioma Inactivated 1 (LGI1) and anti-γ-aminobutyric acid-beta-receptor 1 (GABABR1) has not been published before. CASE PRESENTATION: We herein present the case of a 60-year-old man with slow response, behavioral changes, psychosis and sleep disorders. Laboratory test included serum hyponatremia, positive serum LGI1 and GABABR1 antibodies using transfected cell-based assays. Electroencephalogram exhibited moderate diffusion abnormality. The patient responded well to steroid impulse treatment and sodium supplement therapy, and did not recur during the follow-up. CONCLUSIONS: Here we report the first AE characterized by positive LGI1 and GABABR1 antibodies, as well as summarizing AE with multiple auto-antibodies reported so far, hopefully to provide experience for clinical practice.

RNA m1A Methyltransferase TRMT6 Predicts Poorer Prognosis and Promotes Malignant Behavior in Glioma.

Background: Glioma is the most prevalent central nervous system tumor in humans, and its prognosis remains unsatisfactory due to a lack of effective therapeutic targets. The ectopic expression of N1-methyladenosine (m1A) regulators is a key participant in tumorigenesis and progression. However, the m1A regulator expression status, prognostic value, and relationship with tumor clinical features in glioma remain unclear. Methods: Public datasets were used to analyze the mRNA and protein expression levels of m1A regulators. Kaplan-Meier and Cox regression analyses were performed to confirm the prognostic value of m1A regulators in glioma. Cellular experiments were conducted to verify the effect of TRMT6 on cell function. A comprehensive bioinformatics analysis was conducted to identify the potential molecular mechanisms regulated by TEMT6 in glioma. Results: We found that the dysregulation of m1A regulators was closely associated with tumorigenesis and progression in glioma. Furthermore, TRMT6 might be a powerful and independent biomarker for prognosis in glioma. Our study showed that inhibition of TRMT6 suppressed the proliferation, migration, and invasion of glioma cells. Mechanistically, TRMT6 may be involved in glioma progression by regulating cell cycle, PI3K-AKT, TGF-beta, MTORC1, NOTCH, and MYC pathways. Conclusions: Variation in m1A regulators was closely associated with malignant progression in glioma. Silencing TRMT6 suppressed the cell proliferation, migration, and invasion in glioma. m1A regulators, especially TRMT6, might play an essential role in the malignant progression of glioma.

Observation on the Curative Effect of Microsurgery in 154 Children with Strabismus and Analysis of Its Influencing Factors.

Strabismus is a common ophthalmic disease in the process of child body development, in which the two eyes cannot gaze at the target at the same time, and the incidence of this disease of children is higher. In children with esotropia, exotropia, and up and down strabismus and other typical symptoms, the cause is genetic, innerve, and refractive and regulated, and not receiving timely treatment may lead to stereo vision and diplopia and other phenomena, affecting their learning and life. Surgical treatment is the main treatment for strabismus at present. Traditional orthodontic surgery is performed by doctors under the naked eye, often due to improper operation or suture error and other factors, resulting in more postoperative complications, such as more tissue damage, conjunctival congestion, and muscle suture reaction, which seriously affect the clinical effect of surgical treatment. In recent years, with the continuous development of microsurgical technology, the correction of strabismus under a microscope has been widely carried out in clinic. The operation under the microscope makes the operation more delicate and accurate, overcomes the defects of traditional surgery, and highlights the advantages of minimally invasive surgery. The purpose of this study was to investigate the effect of microsurgical techniques in the treatment of strabismus in children and to analyze the factors influencing the outcome. The results showed that microsurgical strabismus correction in the treatment of strabismus children has short operation time, less intraoperative blood loss, short hospital stay, high efficiency, and less complications, which is worthy of popularization. Age, preoperative strabismus angle, refractive error, distance stereopsis injury, near stereoscopic injury, and duration of disease were all independent influencing factors of postoperative efficacy.

A Novel Method for Food Market Regulation by Emotional Tendencies Predictions from Food Reviews Based on Blockchain and SAEs.

As a part of food safety research, researches on food transactions safety has attracted increasing attention recently. Food choice is an important factor affecting food transactions safety: It can reflect consumer preferences and provide a basis for market regulation. Therefore, this paper proposes a food market regulation method based on blockchain and a deep learning model: Stacked autoencoders (SAEs). Blockchain is used to ensure the fairness of transactions and achieve transparency within the transaction process, thereby reducing the complexity of the trading environment. In order to enhance the usability, relevant Web pages have been developed to make it more friendly and conduct a security analysis for using blockchain. Consumers' reviews after the transactions are finished can be used to train SAEs in order to perform emotional tendencies predictions. Compared with different advanced models for predictions, the test results show that SAEs have a better performance. Furthermore, in order to provide a basis for the formulation of regulation strategies and its related policies, case studies of different traders and commodities have also been conducted, proving the effectiveness of the proposed method.