ABSTRACT
Airway epithelial cells play a crucial role in investigating the physiological and pathological mechanisms of the respiratory tract in yaks, a species whose unique respiratory system has garnered extensive interest. Despite this growing interest, there currently are no available airway epithelial cell lines from yaks, underscoring the crucial need to establish a yak respiratory epithelial cell line. Therefore, our objective was to isolate a population of primary yak nasopharyngeal epithelial cells (pYNE) and transform them into immortalized yak nasopharyngeal epithelial cells (iYNE), assessing their suitability as an in vitro model. Employing a combined method of physical elimination and differential adhesion, we successfully isolated a population of high-purity pYNE, and developed an iYNE line through pCI-neo-hTERT plasmid transfection. Karyotype and transmission electron microscopy analyses confirmed that pYNE and iYNE share identical morphologies and structures. Gel electrophoresis and real-time PCR analyses demonstrated that pYNE and iYNE expressed similar levels of KRT18 and CDH1 genes (p ≥ 0.541). Notably, iYNE expressed a significantly high level of TERT gene expression (p < 0.001). Immunofluorescence analysis demonstrated that both cell types expressed Pan-Cytokeratin, ZO-1, and E-cadherin proteins. Furthermore, immunoblotting analysis indicated significantly higher levels of hTERT and Ki67 proteins in iYNE (p < 0.001), and similar levels of Cluadin-3 and Occludin proteins (p ≥ 0.103). Proliferation curve analysis highlighted iYNE's serum-dependency and significantly enhanced proliferation capacities (p < 0.001). Additionally, pYNE and iYNE cells demonstrated comparable susceptibilities to infectious bovine rhinotracheitis virus (IBRV). These findings collectively suggest that the developed iYNE retains the evaluated physiological characteristics of pYNE, making it an appropriate in vitro model. This advancement will facilitate further investigation into the respiratory physiological and pathological mechanisms in yaks.
ABSTRACT
Breast cancer (BC) is one of the most common fatal cancers. Recent studies have identified the vital role of long noncoding RNA (lncRNAs) in the development and progression of BC. In this research, lncRNA PCAT-1 was studied to identify how it functioned in the metastasis of BC. PCAT-1 expression of tissues was detected by real-time quantitative polymerase chain reaction (RT-qPCR) in 50 BC patients. Cell proliferation, wound healing assay and transwell assay were used to observe the biological behavior changes of BC cells through knockdown or overexpression of PCAT-1. In addition, RT-qPCR and Western blot assay were performed to discover the potential target protein of PCAT-1 in BC. PCAT-1 expression level in BC samples was higher than that of adjacent ones. Besides, cell proliferation, migrated ability and cell invaded ability of BC cells were inhibited after PCAT-1 was silenced. Cell proliferation, migration and invasion of BC cells were promoted after PCAT-1 was overexpressed. In addition, SOX4 was downregulated after silence of PCAT-1 in BC cells, while SOX4 was upregulated after overexpression of PCAT-1 in BC cells. Furthermore, SOX4 was upregulated in BC tissues and was positively associated with PCAT-1. Our study uncovers a new oncogene in BC and suggests that PCAT-1 could enhance BC cell proliferation, migration and invasion via targeting SOX4, which provided a novel therapeutic target for BC patients.
Subject(s)
Breast Neoplasms , Cell Proliferation , Gene Expression Regulation, Neoplastic , Neoplasm Invasiveness , RNA, Long Noncoding , SOXC Transcription Factors , Female , Humans , Breast Neoplasms/pathology , Breast Neoplasms/genetics , Breast Neoplasms/metabolism , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation/genetics , Gene Expression Regulation, Neoplastic/genetics , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , SOXC Transcription Factors/metabolism , SOXC Transcription Factors/geneticsABSTRACT
This study aimed to determine lncRNA SNHG14 and miR-206 in Thyroid Cancer ( TC) and to explore the related mechanisms. Sixty-four samples of thyroid tissue were collected from patients with TC. TC cell lines and a normal human thyroid cell line (HTori-3) were bought. lncRNA SNHG14-siRNA (si-lncRNA SNHG14), lncRNA SNHG14-shRNA (sh-lncRNA SNHG14), blank plasmid (siRNA-NC), miR-206-inhibitor, miR-206-mimics were transfected into BHT101 and Ocut-2C cells. qRT-PCR quantified the expression of lncRNA SNHG14 and miR-206, and the expression of vimentin, Snail, N-cadherin, Slug, E-cadherin and ZO-1 proteins were identified via WB. MTT assay, flow cytometry, and Transwell were employed to determine cellular proliferation, apoptosis, and invasion, separately. The high expression of lncRNA SNHG14 and low expression of miR-206 were exhibited in patients with TC. lncRNA SNHG14 and miR-206 were related to lymph node metastases, TNM staging, as well as differentiation of TC. Silencing lncRNA SNHG14 and over-expressing miR-206 inhibited cell EMT, proliferation, and invasion, but accelerated apoptosis. WB demonstrated that silencing lncRNA SNHG14 and over-expressing miR-206 suppressed the expression of Akt, p-ERK1/2, p-p38, p-4EBP1, p-Akt, PI3K, vimentin, Snail, N-cadherin, and Slugn, as well as up-regulated the expression of E-cadherin and ZO-1. Rescue experiment showed that after BHT101 and Ocut-2C cells were transfected with either sh-lncRNA SNHG14+miR-206-mimics or si-lncRNA SNHG14+miR-206-inhibitor, the cellular proliferative, invasive, and apoptotic activities weren't different from those transfected with siRNA-NC. Suppression of lncRNA SNHG14 up-regulates miR-206 and affects EMT, as well as proliferative, invasive, and apoptotic activities of cells, which may become an underlying treatment target for TC.
Subject(s)
Gene Expression Regulation, Neoplastic , MicroRNAs , RNA, Long Noncoding , Thyroid Neoplasms , Female , Humans , Male , Middle Aged , Apoptosis/genetics , Cell Line, Tumor , Cell Movement , Cell Proliferation , Epithelial-Mesenchymal Transition , Lymphatic Metastasis/genetics , MicroRNAs/genetics , MicroRNAs/metabolism , Neoplasm Invasiveness/genetics , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , Signal Transduction/genetics , Thyroid Neoplasms/genetics , Thyroid Neoplasms/pathology , Thyroid Neoplasms/metabolismABSTRACT
Fusarium oxysporum (FO) is a typical soil-borne pathogenic fungus, and the cucumber wilt disease caused by F. oxysporum f. sp. cucumerinum (FOC) seriously affects crop yield and quality. Vermiculite is increasingly being used as a culture substrate; nevertheless, studies exploring the effectiveness and mechanisms of biocontrol bacteria in this substrate are limited. In this study, vermiculite was used as a culture substrate to investigate the control effect of Bacillus subtilis strain Z-14 on cucumber wilt and the rhizospheric microecology, focusing on colonization ability, soil microbial diversity, and rhizosphere metabolome. Pot experiments showed that Z-14 effectively colonized the cucumber roots, achieving a controlled efficacy of 61.32% for wilt disease. It significantly increased the abundance of Bacillus and the expression of NRPS and PKS genes, while reducing the abundance of FO in the rhizosphere. Microbial diversity sequencing showed that Z-14 reduced the richness and diversity of the rhizosphere bacterial community, increased the richness and diversity of the fungal community, and alleviated the effect of FO on the community structure of the cucumber rhizosphere. The metabolomics analysis revealed that Z-14 affected ABC transporters, amino acid synthesis, and the biosynthesis of plant secondary metabolites. Additionally, Z-14 increased the contents of phenylacetic acid, capsidol, and quinolinic acid, all of which were related to the antagonistic activity in the rhizosphere. Z-14 exhibited a significant control effect on cucumber wilt and influenced the microflora and metabolites in rhizospheric vermiculite, providing a theoretical basis for further understanding the control effect and mechanism of cucumber wilt in different culture substrates.
Subject(s)
Bacillus subtilis , Cucumis sativus , Fusarium , Plant Diseases , Rhizosphere , Soil Microbiology , Fusarium/genetics , Fusarium/physiology , Cucumis sativus/microbiology , Bacillus subtilis/genetics , Bacillus subtilis/physiology , Bacillus subtilis/metabolism , Plant Diseases/microbiology , Plant Diseases/prevention & control , Aluminum Silicates , Plant Roots/microbiologyABSTRACT
BACKGROUND: Thyroid cancer (TC) is a frequent endocrine malignant tumor with various pathologic types. miRNA-363-3p plays a pivotal part in the occurrence, development, prognosis, and treatment of cancer. OBJECTIVE: To explore the mechanism of miRNA-363-3p in TC and provide a new idea for targeted therapy of TC. METHODS: Differential miRNAs and downstream target mRNAs in TC tissues were predicted with bioinformatics analysis. Expression levels of miRNA-363-3p and Synaptotagmin I (SYT1) in TC cells were ascertained by qRT-PCR. Cell migration, invasion, and proliferation were detected by wound healing assay, transwell assay, colony formation assay, CCK-8, and BrdU fluorescence experiment, respectively. Flow cytometry was utilized to detect the levels of apoptosis and necrosis. Immunofluorescence assay was used for detecting autophagosome formation in cells, and the expression levels of autophagy-related proteins, as well as NF-κB related proteins, were measured by western blot. Dual-luciferase reporter gene assay was applied for detecting the interaction between miRNA-363-3p and SYT1. RESULTS: miRNA-363-3p was prominently down-regulated in TC cells. miRNA-363-3p overexpression suppressed migration, invasion, and proliferation, promoting apoptosis and necrosis of TC cells. As the downstream target of miRNA-363-3p, SYT1 was up-regulated in TC cells. SYT1 overexpression reversed the inhibition of TC cell proliferation, invasion, migration, and autophagy mediated by miRNA-363-3p overexpression. In addition, miRNA-363-3p overexpression inhibited the activation of the NF-κB pathway in cells, while further overexpression of SYT1 weakened the inhibition of miRNA-363-3p overexpression on the NF-κB pathway. CONCLUSION: miRNA-363-3p affected the NF-κB signaling pathway by down-regulating SYT1 expression to inhibit the malignant progression of TC cells, providing theoretical support for the treatment of TC.
Subject(s)
MicroRNAs , Thyroid Neoplasms , Humans , Apoptosis/genetics , Autophagy/genetics , Cell Line, Tumor , Cell Proliferation/genetics , MicroRNAs/metabolism , Necrosis , NF-kappa B/metabolism , Synaptotagmin I , Thyroid Neoplasms/genetics , Thyroid Neoplasms/pathologyABSTRACT
Continuous cropping of the same crop leads to soil degradation and a decline in crop production, and these impacts could be mitigated through rotation cropping. Although crop rotation enhances soil fertility, microbial community diversity, and potato yield, its effects on the soil ecosystem multifunctionality (EMF) remain unclear. In the present research, we comparatively examined the effects of potato continuous cropping (PP) and rotation cropping [potato-oat rotation (PO) and potato-forage maize rotation (PFM)] on the soil EMF as well as the roles of keystone taxa, microbes abundance, and chemical properties in EMF improvement. It was demonstrated that soil EMF is increased in rotation cropping (PO and PFM) than PP. Soil pH was higher in rotation cropping (PO and PFM) than in PP, while total phosphorus (TP) and available phosphorus (AP) were significantly decreased than that in PP. Rotation cropping (PO and PFM) markedly changed the bacterial and fungal community compositions, and improved the potential plant-beneficial fungi, e.g., Schizothecium and Chaetomium, while reducing the abundances of the potentially phytopathogenic fungi, e.g., Alternaria, Fusarium, Verticillium dahiae, Gibberella, Plectosphaerella, Colletotrichum, Phoma, and Lectera in comparison with PP. Also, co-occurrence patterns for bacteria and fungi were impacted by crop rotation, and keystone taxa, e.g., Nitrospira.1, Lysinibacillus, Microlunatus.1, Sphingomonas.3, Bryobacter.1, Micromonospora, and Schizothecium, were enriched in PO and PFM than PP. The structural equation model (SEM) further demonstrated that cropping systems increased soil ecosystem multifunctionality through regulating SOM and keystone taxa (Schizothecium1), and keystone taxa were mediated by soil pH. This study suggested that rotation cropping might contribute to the improvement of soil ecosystem multifunctionality as well as the development of disease-suppressive soils in comparison with potato continuous cropping.
ABSTRACT
Objective: The aim of the study is to investigate the effect of modified nonpneumatic transaxillary approach in the treatment of thyroid cancer and its effect on immune function and parathyroid function. Methods: A total of 96 patients with thyroid cancer who were diagnosed and treated in our hospital from January 2018 to December 2020 were selected and randomly divided into the control group of 48 cases and the observation group of 48 cases. The control group was given open surgery, and for the observation group, modified nonpneumatic transaxillary approach was used for treatment. The perioperative related indicators, the incidence of complications, as well as the changes of immune function indicators, parathyroid hormone (PTH), and calcium before and after surgery were compared between the two groups. Results: The time of flap separation and cavity construction, operation time, and hospital stay in the observation group were significantly longer than those in the control group (P < 0.05). After operation, CD3+, CD4+, and CD4+/CD8+ in the two groups were lower than those before operation (P < 0.05), but the observation group was significantly higher than that in the control group (P < 0.05). The serum PTH and calcium at 1 h, 1 d, 3 d and 7 d after operation were lower than those before operation in this group (P < 0.05), but the observation group was significantly higher than that in the control group (P < 0.05). Compared with the control group, the incidence of complications in the observation group (4.17% vs. 6.25%) was not statistically significant (P > 0.05). Conclusion: Compared with open surgery, the modified nonpneumatic transaxillary approach in the treatment of thyroid cancer is more effective in reducing immune function decline, hypoparathyroidism, and hypocalcemia; although the operation time and recovery time are longer, and it is safe. Sex is also high.
ABSTRACT
Ketone bodies are crucial intermediate metabolites widely associated with treating metabolic diseases. Accumulating evidence suggests that ketone bodies may act as immunoregulators in humans and animals to attenuate pathological inflammation through multiple strategies. Although the clues are scattered and untrimmed, the elevation of these ketone bodies in the circulation system and tissues induced by ketogenic diets was reported to affect the immunological barriers, an important part of innate immunity. Therefore, beta-hydroxybutyrate, a key ketone body, might also play a vital role in regulating the barrier immune systems. In this review, we retrospected the endogenous ketogenesis in animals and the dual roles of ketone bodies as energy carriers and signal molecules focusing on beta-hydroxybutyrate. In addition, the research regarding the effects of beta-hydroxybutyrate on the function of the immunological barrier, mainly on the microbiota, chemical, and physical barriers of the mucosa, were outlined and discussed. As an inducible endogenous metabolic small molecule, beta-hydroxybutyrate deserves delicate investigations focusing on its immunometabolic efficacy. Comprehending the connection between ketone bodies and the barrier immunological function and its underlining mechanisms may help exploit individualised approaches to treat various mucosa or skin-related diseases.
Subject(s)
Diet, Ketogenic , Ketone Bodies , 3-Hydroxybutyric Acid , Animals , Immunity, Innate , InflammationABSTRACT
Abnormal glycemia is frequently along with nephritis, whose pathogenesis is unexplicit. Here, we investigated the effects of abnormal glucose on the renal glomerulus epithelial cells by stimulating immortalized bovine renal glomerulus epithelial (MDBK) cells with five different levels of glucose, including low glucose (2.5 mM for 48 h, LG), normal glucose (5 mM for 48 h, NG), high glucose (25 mM for 48 h, HG), increasing glucose (24 h of 2.5 mM glucose followed by 24 h of 25 mM, IG), and reducing glucose (24 h of 25 mM glucose followed by 24 h of 2.5 mM, RG). The results showed that LG and RG treatments had nonsignificant effects (p > 0.05) on the viability of MDBK cells. HG treatment decreased the viabilities of cells (p < 0.01) without triggering an apparent inflammatory response by activating the nox4/ROS/p53/caspase-3-mediated apoptosis pathway. IG treatment decreased the viabilities of cells significantly (p < 0.01) with high levels of pro-inflammatory cytokines IL-1ß and IL-18 in the supernatant (p < 0.05) by triggering the txnip/nlrp3/gsdmd-mediated pyroptosis pathway. These results indicated that the process of glucose increase rather than the constant high glucose was the main cause of abnormal glucose-induced MDBK cell inflammatory death, prompting that the process of glycemia increases might be mainly responsible for the nephritis in diabetic nephropathy, underlining the importance of glycemic control in diabetes patients.
Subject(s)
Diabetic Nephropathies , Nephritis , Humans , Animals , Cattle , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Inflammasomes/metabolism , Glucose/metabolism , Diabetic Nephropathies/metabolism , Epithelial Cells/metabolism , PyroptosisABSTRACT
OBJECTIVE: To ascertain the mechanism of miRNA-144-5p and ITGA3 in thyroid cancer (TC). METHODS: From The Cancer Genome Atlas (TCGA), RNA expression profiles were obtained for the expression analysis of miRNAs and mRNAs in TC. qRT-PCR and western blot were utilized to measure the expression of miRNA-144-5p and ITGA3 at RNA and protein levels, respectively. The association between miRNA-144-5p and ITGA3 was validated by the dual-luciferase assay. CCK-8, scratch healing, transwell, and flow cytometry assays were employed to evaluate tumor-related cell behaviors. RESULTS: Low-expressed miRNA-144-5p and high-expressed ITGA3 were found in TC cells relative to normal cells. miRNA-144-5p expression was positively associated with suppressive effects on proliferative, invasive, and migratory ability of TC cells while negatively associated with cell apoptosis. miRNA-144-5p inhibited ITGA3 expression in TC, and its overexpression remarkably reversed the tumor-promoting effects of overexpressed ITGA3 on the biological functions of TC. CONCLUSION: It is verified in our study that cell growth of TC is inhibited by the miRNA-144-5p/ITGA3 axis, which represents an underlying target for TC. This research proposed a new insight into the strategy of TC treatment.
Subject(s)
Integrin alpha3/genetics , MicroRNAs/genetics , Thyroid Neoplasms/genetics , Apoptosis/genetics , Carcinogens/metabolism , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation/genetics , Computational Biology , Disease Progression , Down-Regulation , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Humans , Integrin alpha3/metabolism , MicroRNAs/metabolism , Neoplasm Invasiveness/genetics , Thyroid Neoplasms/metabolism , Thyroid Neoplasms/pathologyABSTRACT
In response to the concept of "compact energy storage", research on electrolyte dosage dwindling is definitely efficient owing to present electrolyte usage up to 70 wt % in a cell. While less electrolyte usage leads to slow reaction kinetics. Herein, a heterojunction, MoP/MoS2 core with much defects and vacancies coated by porous carbon shell, is synthesized. Besides, the small particle size of MoP/MoS2@C facilitates a close packing to form a dense and porous modified layer on PP-based (F-PP) separator. The heterojunction with defects exposes abundant interfaces and assures an adequate local electrolyte availability and an improved electrolyte affinity that are beneficial for Li+ transfer. When using F-PP separator, Li-S cell performs well in the lean electrolyte. Apart from a high discharging capacity of 517.1 mAh g-1 at 5 C in E/S = 10 (only half benchmark dosage), the cell realizes a favorable stability at C/2 over 500 cycles even in E/S = 7 (0.065% decay per cycle), demonstrating an effective polysulfides (PS) shuttling relief and reversibility of PS-relating chemical conversion. All these enhanced electrochemical behaviors in lean electrolyte result from a three-in-one strategy realized by defects-included MoP/MoS2@C heterojunction, including incorporating the lithiuphilic and sulfophilic sites for PS confinement and electrocatalysis triggered by abundant S vacancies and Lewis and Brønsted acid sites.
ABSTRACT
BACKGROUND: The development of low-carbon agriculture is promising for mitigating climate change. This study used adjustments to the planting structure in Zhangbei County, China, as an example to evaluate whether the carbon footprint per unit of economic benefit is a suitable indicator of low-carbon agriculture and to determine if low-carbon agriculture is not necessarily low-input non-intensive agriculture. RESULTS: The results showed that total greenhouse gas emissions increased; therefore, the adjustments to the planting structure were ostensibly not a low-carbon process. However, if we obtain the same economic benefit as the actual distribution of the planting industry by adopting the scenario of planting only grain crops, then the annual greenhouse gas emissions would be 1608.00 × 103 t CO2 eq, and 5769.94 × 103 ha of farmland would be required. However, if we adopt the scenario of planting only vegetable crops, then only 82.52 × 103 ha of farmland would be required, and the annual greenhouse gas emissions would be 323.52 × 103 t CO2 eq. CONCLUSIONS: These results indicated that the carbon footprint per unit of economic benefit is a suitable indicator to assess agricultural sustainability and that intensive agriculture with high input and high output is a form of low-carbon agriculture if the carbon footprint per unit of economic benefit is low. © 2019 Society of Chemical Industry.
Subject(s)
Agriculture/economics , Agriculture/methods , Carbon Footprint/economics , Carbon , China , Cost-Benefit Analysis , Crops, Agricultural/growth & development , Edible Grain/growth & development , Farms , Greenhouse Gases/metabolism , Sustainable Development , Vegetables/growth & developmentABSTRACT
Optimizing the nitrogen (N) application rate can increase crop yield while reducing the environmental risks. However, the optimal N rates vary substantially when different targets such as maximum yield or maximum economic benefit are considered. Taking the wheat-maize rotation cropping system on the North China Plain as a case study, we quantified the variation of N application rates when targeting constraints on yield, economic performance, N uptake and N utilization, by conducting field experiments between 2011 and 2013. Results showed that the optimal N application rate was highest when targeting N uptake (240kgha-1 for maize, and 326kgha-1 for wheat), followed by crop yield (208kgha-1 for maize, and 277kgha-1 for wheat) and economic income (191kgha-1 for maize, and 253kgha-1 for wheat). If environmental costs were considered, the optimal N application rates were further reduced by 20-30% compared to those when targeting maximum economic income. However, the optimal N rate, with environmental cost included, may result in soil nutrient mining under maize, and an extra input of 43kgNha-1 was needed to make the soil N balanced and maintain soil fertility in the long term. To obtain a win-win situation for both yield and environment, the optimal N rate should be controlled at 179kgha-1 for maize, which could achieve above 99.5% of maximum yield and have a favorable N balance, and at 202kgha-1 for wheat to achieve 97.4% of maximum yield, which was about 20kgNha-1 higher than that when N surplus was nil. Although these optimal N rates vary on spatial and temporal scales, they are still effective for the North China Plain where 32% of China's total maize and 45% of China's total wheat are produced. More experiments are still needed to determine the optimal N application rates in other regions. Use of these different optimal N rates would contribute to improving the sustainability of agricultural development in China.
ABSTRACT
AIM: The aims of this study are to identify which type of tooth has the strong relationship between age and pulp cavity/chamber volume among 13 types of tooth from the same dentition and to determine whether the inclusion of multiple types of tooth may improve the accuracy of age estimation. MATERIALS AND METHODS: Cone beam computed tomography (CBCT) images from 115 females and 125 males aged between 16 and 63 years were analyzed. The DICOM data of all the images were imported into ITK-SNAP 2.4 for the calculation of pulp cavity/chamber volumes. Logarithmic regression analysis and multiple linear regression analysis were applied to establish the relationship between age and pulp cavity/chamber volumes. RESULTS: Among the 13 types of tooth, maxillary second molars have the largest R (2) (0.491, 0.642, and 0.498) and the smallest SEE (8.119, 6.754, and 8.022) in male, female, and pooled gender samples, respectively. The multiple linear regression analysis for the combination of multi-types of tooth indicated that a larger R (2) (0.627, 0.701, and 0.631) and smaller SEE (7.100, 6.258, and 6.970) than the counterpart calculated from the logarithmic regression analysis of a single type of tooth in male, female, and pooled gender samples, respectively. CONCLUSION: The pulp chamber volume of the maxillary second molars has the largest correlation coefficient with age. Using multiple types of tooth may improve the accuracy of age estimation compared with only one type of tooth used.
Subject(s)
Age Determination by Teeth/methods , Cone-Beam Computed Tomography , Dental Pulp/anatomy & histology , Dental Pulp/diagnostic imaging , Adolescent , Adult , Female , Humans , Linear Models , Male , Middle Aged , Retrospective Studies , Young AdultABSTRACT
A highly sensitive and ultrasensitive electrochemical aptasensor for platelet-derived growth factor BB (PDGF-BB) detection is fabricated based on layered molybdenum selenide-graphene (MoSe2-Gr) composites and Exonuclease III (Exo III)-aided signal amplification. MoSe2-Gr is prepared by a simple hydrothermal method and used as a promising sensing platform. Exo III has a specifical exo-deoxyribonuclease activity for duplex DNAs in the direction from 3' to 5' terminus, however its activity is limited on the duplex DNAs with more than 4 mismatched terminal bases at 3' ends. Herein, aptamer and complementary DNA (cDNA) sequences are designed with four thymine bases on 3' ends. In the presence of target protein, the aptamer associates with it and facilitates the formation of duplex DNA between cDNA and signal DNA. The duplex DNA then is digested by Exo III and releases cDNA, which hybridizes with signal DNA to perform a new cleavage process. Nevertheless, in the absence of target protein, the aptamer hybridizes with cDNA will inhibit the Exo III-assisted nucleotides cleavage. The signal DNA then hybridizes with capture DNA on the electrode. Subsequently, horse radish peroxidase is fixed on electrode by avidin-biotin reaction and then catalyzes hydrogen peroxide and hydroquinone to produce electrochemical response. Therefore, a bridge can be established between the concentration of target protein and the degree of the attenuation of the obtained signal, providing a quantitative measure of target protein with a broad detection range of 0.0001-1 nM and a detection limit of 20 fM.
Subject(s)
Aptamers, Nucleotide/chemistry , Biosensing Techniques , Conductometry/instrumentation , Exodeoxyribonucleases/chemistry , Microchemistry/instrumentation , Nanocomposites/chemistry , Equipment Design , Equipment Failure Analysis , Graphite/chemistry , Metal Nanoparticles/chemistry , Metal Nanoparticles/ultrastructure , Molybdenum/chemistry , Nanocomposites/ultrastructure , Nucleic Acid Amplification Techniques/instrumentation , Platelet-Derived Growth Factor/chemistry , Selenium/chemistryABSTRACT
Intercropping is regarded as an important agricultural practice to improve crop production and environmental quality in the regions with intensive agricultural production, e.g., northern China. To optimize agronomic advantage of maize (Zea mays L.) and soybean (Glycine max L.) intercropping system compared to monoculture of maize, two sequential experiments were conducted. Experiment 1 was to screening the optimal cropping system in summer that had the highest yields and economic benefits, and Experiment 2 was to identify the optimum row ratio of the intercrops selected from Experiment 1. Results of Experiment 1 showed that maize intercropping with soybean (maize || soybean) was the optimal cropping system in summer. Compared to conventional monoculture of maize, maize || soybean had significant advantage in yield, economy, land utilization ratio and reducing soil nitrate nitrogen (N) accumulation, as well as better residual effect on the subsequent wheat (Triticum aestivum L.) crop. Experiment 2 showed that intercropping systems reduced use of N fertilizer per unit land area and increased relative biomass of intercropped maize, due to promoted photosynthetic efficiency of border rows and N utilization during symbiotic period. Intercropping advantage began to emerge at tasseling stage after N topdressing for maize. Among all treatments with different row ratios, alternating four maize rows with six soybean rows (4M:6S) had the largest land equivalent ratio (1.30), total N accumulation in crops (258 kg ha(-1)), and economic benefit (3,408 USD ha(-1)). Compared to maize monoculture, 4M:6S had significantly lower nitrate-N accumulation in soil both after harvest of maize and after harvest of the subsequent wheat, but it did not decrease yield of wheat. The most important advantage of 4M:6S was to increase biomass of intercropped maize and soybean, which further led to the increase of total N accumulation by crops as well as economic benefit. In conclusion, alternating four maize rows with six soybean rows was the optimum row ratio in maize || soybean system, though this needs to be further confirmed by pluri-annual trials.
Subject(s)
Glycine max/growth & development , Triticum/growth & development , Zea mays/growth & development , Agriculture/methods , Biomass , China , Crops, Agricultural/economics , Crops, Agricultural/growth & development , SeasonsABSTRACT
AIM: To establish a method that can be used for human age estimation on the basis of pulp chamber volume of first molars and to identify whether the method is good enough for age estimation in real human cases. MATERIALS AND METHODS: CBCT images of 373 maxillary first molars and 372 mandibular first molars were collected to establish the mathematical model from 190 female and 213 male patients whose age between 12 and 69 years old. The inclusion criteria of the first molars were: no caries, no excessive tooth wear, no dental restorations, no artifacts due to metal restorative materials present in adjacent teeth, and no pulpal calcification. All the CBCT images were acquired with a CBCT unit NewTom VG (Quantitative Radiology, Verona, Italy) and reconstructed with a voxel-size of 0.15mm. The images were subsequently exported as DICOM data sets and imported into an open source 3D image semi-automatic segmenting and voxel-counting software ITK-SNAP 2.4 for the calculation of pulp chamber volumes. A logarithmic regression analysis was conducted with age as dependent variable and pulp chamber volume as independent variables to establish a mathematical model for the human age estimation. To identify the precision and accuracy of the model for human age estimation, another 104 maxillary first molars and 103 mandibular first molars from 55 female and 57 male patients whose age between 12 and 67 years old were collected, too. Mean absolute error and root mean square error between the actual age and estimated age were used to determine the precision and accuracy of the mathematical model. The study was approved by the Institutional Review Board of Peking University School and Hospital of Stomatology. RESULTS: A mathematical model was suggested for: AGE=117.691-26.442×ln (pulp chamber volume). The regression was statistically significant (p=0.000<0.01). The coefficient of determination (R(2)) was 0.564. There is a mean absolute error of 8.122 and root mean square error of 5.603 between the actual age and estimated age for all the tested teeth. CONCLUSION: The pulp chamber volume of first molar is a useful index for the estimation of human age with reasonable precision and accuracy.
Subject(s)
Age Determination by Teeth/methods , Cone-Beam Computed Tomography , Dental Pulp/diagnostic imaging , Molar/diagnostic imaging , Adolescent , Adult , Aged , Child , China , Female , Forensic Dentistry , Humans , Imaging, Three-Dimensional , Male , Middle Aged , Models, Biological , Young AdultABSTRACT
The aerobic oxidation of pyrrolones catalyzed by Fe(OTf)3 to form reactive N-acyliminium ion intermediates that undergo nucleophilic additions with alcohols to give the corresponding products in moderate to good yields is described.
Subject(s)
Iron/chemistry , Pyrroles/chemistry , Catalysis , Oxidation-Reduction , Oxygen/chemistryABSTRACT
The multi-channel visible bremsstrahlung measurement system has been developed on Experimental Advanced Superconducting Tokamak (EAST). In addition to providing effective ion charge Zeff as a routine diagnostic, this diagnostic can also be used to estimate other parameters. With the assumption that Zeff can be seen as constant across the radius and does not change significantly during steady state discharges, central electron temperature, averaged electron density, electron density profile, and plasma current density profile have been obtained based on the scaling of Zeff with electron density and the relations between Zeff and these parameters. The estimated results are in good coincidence with measured values, providing an effective and convenient method to estimate other plasma parameters.
ABSTRACT
We have developed a sensitive sensing platform for 17ß-estradiol by combining the aptamer probe and hybridization reaction. In this assay, 2-dimensional cobalt sulfide nanosheet (CoS) was synthesized by a simple hydrothermal method with L-cysteine as sulfur donor. An electrochemical aptamer biosensor was constructed by assembling a thiol group tagged 17ß-estradiol aptamer on CoS and gold nanoparticles (AuNPs) modified electrode. Methylene blue was applied as a tracer and a guanine-rich complementary DNA sequence was designed to bind with the unbound 17ß-estradiol aptamer for signal amplification. The binding of guanine-rich DNA to the aptamer was inhibited when the aptamer captured 17ß-estradiol. Using guanine-rich DNA in the assay greatly amplified the redox signal of methylene blue bound to the detection probe. The CoS/AuNPs film formed on the biosensor surface appeared to be a good conductor for accelerating the electron transfer. The method demonstrated a high sensitivity of detection with the dynamic concentration range spanning from 1.0×10(-9) to 1.0×10(-12) M and a detection limit of 7.0×10(-13) M. Besides, the fabricated biosensor exhibited good selectivity toward 17ß-estradiol even when interferents were presented at 100-fold concentrations. Our attempt will extend the application of the CoS nanosheet and this signal amplification assay to biosensing areas.