What is the stable internal fixation for the unstable and osteoporotic supracondylar femoral fractures: a finite element analysis.

BACKGROUND: Osteoporotic supracondylar femoral fractures (OSFF) have historically been managed by the lateral anatomical locking plate with reasonable success. However, for some kinds of unstable and osteoporotic supracondylar femoral fractures (UOSFF), especially with bone defects, unilateral locking plate (ULLP) fixation failed or resulted in implant breakage. This paper is going to explore what is the stable internal fixation for UOSFF by adding the bilateral locking plate (BLLP) fixation. METHODS: OSFF models were divided into two groups according to the fracture line type, which would be further subdivided according to their angle of fracture line, presence of bone defect, location, and degree of bone defect. Thereafter, kinds of locking plate fixation were constructed. A 2010-N load was applied to the femoral head, and a 1086-N load was applied to the greater trochanter. In this condition, the maximum von Mises stress distribution of models were investigated. RESULTS: Firstly, it was obviously found that the stress concentration in the BLLP group was more dispersed than that in the ULLP group. Secondly, according to the fracture line analysis, the stress value of fracture line type in "\" model group was higher than that of "/" model group. Moreover, with the increase in fracture line angle, the stress value of the model increased. Thirdly, from the bone defect analysis, the stress value of the medial bone defect (MBD) model group was higher than that of the lateral bone defect (LBD) model group. And as the degree of bone defect increased, the stress value increased gradually in the model group. CONCLUSION: In the following four cases, lateral unilateral locking plate fixation cannot effectively stabilize the fracture end, and double locking plate internal fixation is a necessary choice. First, when the angle of the fracture line is large (30, 45). Second, when the fracture line type is "/." Third, when the bone defect is large. Fourth, when the bone defect is medial.

Development of a Comprehensive Quality Evaluation System for Foxtail Millet from Different Ecological Regions.

Foxtail millet (Setaria italica L.) is a critical grain with high nutritional value and the potential for increased production in arid and semiarid regions. The foxtail millet value chain can be upgraded only by ensuring its comprehensive quality. Thus, samples were collected from different production areas in Shanxi province, China, and compared in terms of quality traits. We established a quality evaluation system utilizing multivariate statistical analysis. The results showed that the appearance, nutritional content, and culinary value of foxtail millet produced in different ecological regions varied substantially. Different values of amino acids (DVAACs), alkali digestion values (ADVs), and total flavone content (TFC) had the highest coefficients of variation (CVs) of 50.30%, 39.75%, and 35.39%, respectively. Based on this, a comprehensive quality evaluation system for foxtail millet was established, and the quality of foxtail millet produced in the five production areas was ranked in order from highest to lowest: Dingxiang > Zezhou > Qinxian > Xingxian > Yuci. In conclusion, the ecological conditions of Xinding Basin are favorable for ensuring the comprehensive quality of foxtail millet. .

Transcriptome Analysis Reveals Brassinolide Signaling Pathway Control of Foxtail Millet Seedling Starch and Sucrose Metabolism under Freezing Stress, with Implications for Growth and Development.

Low-temperature stress limits the growth and development of foxtail millet. Freezing stress caused by sudden temperature drops, such as late-spring coldness, often occurs in the seedling stage of foxtail millet. However, the ability and coping strategies of foxtail millet to cope with such stress are not clear. In the present study, we analyzed the self-regulatory mechanisms of freezing stress in foxtail millet. We conducted a physiological study on foxtail millet leaves at -4 °C for seven different durations (0, 2, 4, 6, 8, 10, and 12 h). Longer freezing time increased cell-membrane damage, relative conductance, and malondialdehyde content. This led to osmotic stress in the leaves, which triggered an increase in free proline, soluble sugar, and soluble protein contents. The increases in these substances helped to reduce the damage caused by stress. The activities of superoxide dismutase, peroxidase, and catalase increased reactive oxygen species (ROS) content. The optimal time point for the response to freezing stress was 8 h after exposure. The transcriptome analysis of samples held for 8 h at -4 °C revealed 6862 differentially expressed genes (DEGs), among which the majority are implicated in various pathways, including the starch and sucrose metabolic pathways, antioxidant enzyme pathways, brassinolide (BR) signaling pathway, and transcription factors, according to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment. We investigated possible crosstalk between BR signals and other pathways and found that BR signaling molecules were induced in response to freezing stress. The beta-amylase (BAM) starch hydrolase signal was enhanced by the BR signal, resulting in the accelerated degradation of starch and the formation of sugars, which served as emerging ROS scavengers and osmoregulators to resist freezing stress. In conclusion, crosstalk between BR signal transduction, and both starch and sucrose metabolism under freezing stress provides a new perspective for improving freezing resistance in foxtail millet.

The relationship between ecological factors and commercial quality of high-quality foxtail millet "Jingu 21".

The commercial quality of foxtail millet grain (Setaria italica L.) includes appearance quality, functional quality, and cooking and eating quality, which directly determine whether consumers will purchase the product. We studied the relationship between ecological factors and commercial quality attributes of foxtail millet "Jingu 21" from twelve production areas. The results showed that altitude, latitude, and diurnal temperature range were negatively correlated with b*, total flavones content (TFC), setback (SB), consistence (CS) and pasting temperature (PTM), but positively correlated with L/B and breakdown (BD). In contrast, average temperature, average precipitation, average humidity, available nitrogen, phosphorus, and potassium had positive effects on 1,000-grain weight (KGW), b*, TFC, CS, and PTM and had a negative impact on L/B and BD. Climate factors had a greater effect on the commercial quality of foxtail millet than soil factors, and the influence of climatic factors was particularly obvious in the early and middle growth periods. The multivariate equation between ecological factors and the comprehensive score of foxtail millet commercial quality is Y = 1,159.745-4.496X1 (altitude) + 19.529X5 (≥10℃ effective accumulated temperature) - 166.327X10 (organic matters). In conclusion, high temperature and precipitation are conducive to high quality appearance and the accumulation of functional substances, while a high diurnal temperature range and high soil nutrients are conducive to the formation of cooking and eating quality. The impact of ecological factors on foxtail millet quality is complicated and it is essential to select a cultivation site that is matched to the intended use of the foxtail millet being produced.

Low copy numbers for mitochondrial DNA moderates the strength of nuclear-cytoplasmic incompatibility in plants.

Plant cells contain only small amounts of mitochondrial DNA (mtDNA), with the genomic information shared among multiple mitochondria. The biological relevance and molecular mechanism underlying this hallmark of plant cells has been unclear. Here, we report that Arabidopsis thaliana plants exhibited significantly reduced growth and mitochondrial dysfunction when the mtDNA copy number was increased to the degree that each mitochondrion possessed DNA. The amounts of mitochondrion-encoded transcripts increased several fold in the presence of elevated mtDNA levels. However, the efficiency of RNA editing decreased with this excess of mitochondrion-encoded transcripts, resulting in impaired assembly of mitochondrial complexes containing mtDNA-encoded subunits, such as respiratory complexes I and IV. These observations indicate the occurrence of nuclear-mitochondrial incompatibility in the cells with increased amounts of mtDNA and provide an initial answer to the fundamental question of why plant cells have much lower mtDNA levels than animal cells. We propose that keeping mtDNA levels low moderates nuclear-mitochondrial incompatibility and that this may be a crucial factor driving plant cells to restrict the copy numbers of mtDNA.

Distinctive physiological and molecular responses of foxtail millet and maize to nicosulfuron.

Introduction: Nicosulfuron is the leading acetolactate synthase inhibitor herbicide product, and widely used to control gramineous weeds. Here, we investigated the metabolic process of nicosulfuron into foxtail millet and maize, in order to clarify the mechanism of the difference in sensitivity of foxtail millet and maize to nicosulfuron from the perspective of physiological metabolism and provide a theoretical basis for the breeding of nicosulfuron-resistant foxtail millet varieties. Methods: We treated foxtail millet (Zhangzagu 10, Jingu 21) and maize (Nongda 108, Ditian 8) with various doses of nicosulfuron in both pot and field experiments. The malonaldehyde (MDA) content, target enzymes, detoxification enzymes, and antioxidant enzymes, as well as related gene expression levels in the leaf tissues of foxtail millet and maize were measured, and the yield was determined after maturity. Results: The results showed that the recommended dose of nicosulfuron caused Zhangzagu 10 and Jingu 21 to fail to harvest; the yield of the sensitive maize variety (Ditian 8) decreased by 37.09%, whereas that of the resistant maize variety (Nongda 108) did not decrease. Nicosulfuron stress increased the CYP450 enzyme activity, MDA content, and antioxidant enzyme activity of foxtail millet and maize, reduced the acetolactate synthase (ALS) activity and ALS gene expression of foxtail millet and Ditian 8, and reduced the glutathione S-transferase (GST) activity and GST gene expression of foxtail millet. In conclusion, target enzymes, detoxification enzymes, and antioxidant enzymes were involved in the detoxification metabolism of nicosulfuron in plants. ALS and GST are the main factors responsible for the metabolic differences among foxtail millet, sensitive maize varieties, and resistant maize varieties. Discussion: These findings offer valuable insights for exploring the target resistance (TSR) and non-target resistance (NTSR) mechanisms in foxtail millet under herbicide stress and provides theoretical basis for future research of develop foxtail millet germplasm with diverse herbicide resistance traits.

Effects of Basal Selenium Fertilizer Application on Agronomic Traits, Yield, Quality, and Se Content of Dryland Maize.

To explore the efficiency of selenium (Se) fertilizer application in dryland maize, we tested five Se fertilizer application treatments: 0 g ha-1 (Se0), 75 g ha-1 (Se1), 150 g ha-1 (Se2), 225 g ha-1 (Se3), and 300 g ha-1 (Se4). Compared with Se0, Se2 increased the leaf area, chlorophyll content, internode length, plant height, and ear height by 7.95%, 3.20%, 13.19%, 1.89%, and 7.98%, respectively. Se2 and Se3 significantly affected the stem internode diameter, cortex thickness, and cellulose content, which were positively correlated with lodging resistance. Compared with Se0, Se3 promoted the contents of soluble sugar, crude protein, crude fat, and starch in grains, which increased by 9.48%, 6.59%, 1.56%, and 4.82%, respectively. It implies that maize grain application of Se significantly improves their Se content. Se1 did not observably influence the growth of maize, and the promoting effect of Se4 on maize decreased. The lodging resistance of maize as analyzed by Pearson correlation analysis correlated with the application of Se fertilizer. It proved that higher yield, grain quality, grain Se content, and lodging resistance of stems were concerned with Se fertilizer application in the range of 150-225 g ha-1. The results provide useful information for Se fertilizer treatment in dryland maize.

Effect of Pyrazosulfuron-Methyl on the Photosynthetic Characteristics and Antioxidant Systems of Foxtail Millet.

Foxtail millet (Setaria Italica L.) plays a principal role in food security in Africa and Asia, but it is sensitive to a variety of herbicides. This study was performed to clarify whether pyrazosulfuron-methyl can be used in foxtail millet fields and the effect of pyrazosulfuron-methyl on the photosynthetic performance of foxtail millet. Two foxtail millet varieties (Jingu 21 and Zhangzagu 10) were subjected to five doses (0, 15, 30, 60, and 120 g ai ha-1) of pyrazosulfuron-methyl in pot and field experiments. The plant height, leaf area, stem diameter, photosynthetic pigment contents, gas exchange parameters, chlorophyll fluorescence parameters, antioxidant enzyme activities, and antioxidant contents at 7 and 15 days after pyrazosulfuron-methyl application, and the yield of foxtail millet were measured. The results suggested that pyrazosulfuron-methyl inhibited the growth of foxtail millet and reduced the photosynthetic pigment contents, photosynthetic rate, and photosynthetic system II activity. Similarly, pyrazosulfuron-methyl decreased the antioxidant enzyme activities and antioxidant contents. These results also indicated that the toxicity of pyrazosulfuron-methyl to foxtail millet was decreased gradually with the extension of time after application; however, the foxtail millet yield was still significantly reduced. Therefore, pyrazosulfuron-methyl is not recommended for application in foxtail millet fields.

Polysaccharide from wild morels alters the spatial structure of gut microbiota and the production of short-chain fatty acids in mice.

Polysaccharides from morels possess many characteristics beneficial to health, such as anti-tumor and immunomodulatory activities. The gut microbiota plays a critical role in the modulation of immune function. However, the impact of morel polysaccharides on the gut microbiota has not yet been explored. In this study, a high-throughput pyrosequencing technique was used to investigate the effects of MP, a new heteropolysaccharide extracted from wild morels, on the diversity and composition of microbiota along the intestine in mice, as well as the production of short-chain fatty acids (SCFAs). The results showed that MP treatment increased the number of operational taxonomic unit (OTUs) and diversity along the intestine, especially in the small intestine. MP treatment induced a significant decrease in the number of Firmicutes and a significant increase in the number of Bacteroidetes in the small intestine microbiota. It was also observed that the relative abundance of SCFA-producing bacteria, especially Lachnospiraceae, was increased in both the cecum and colon of MP-treated mice. Moreover, MP promoted the production of SCFAs in mice. These results provide a foundation for further understanding the health benefits conferred by morel polysaccharides.

Effects of polysaccharides from wild morels on immune response and gut microbiota composition in non-treated and cyclophosphamide-treated mice.

Polysaccharides isolated from mushrooms have been identified as potential prebiotics that could impact gut microbiota. In this study, a water-soluble polysaccharide (MP) extracted from wild morels was evaluated for its effects on the gut microbiota of non-treated and cyclophosphamide (CP)-treated mice. The results showed that MP restored the spleen weight and increased the counts of white blood cells and lymphocytes in the peripheral blood and spleen of the CP-treated mice. Mice treated with MP exhibited increased levels of short-chain fatty acid (SCFA)-producing bacteria, especially Lachnospiraceae, compared to normal mice, and increased levels of Bacteroidetes and SCFA-producing bacteria, especially Ruminococcaceae, compared to the CP-treated mice. Moreover, MP treatment increased the production of valeric acid and decreased the production of acetic acid in the non-treated mice and increased the production of acetic acid, propionic acid, butyric acid, and valeric acid in the CP-treated mice. These results show that MP is potentially good for health.

Preparation and Characterization of Edible Dialdehyde Carboxymethyl Cellulose Crosslinked Feather Keratin Films for Food Packaging.

The development of edible films based on the natural biopolymer feather keratin (FK) from poultry feathers is of great interest to food packaging. Edible dialdehyde carboxymethyl cellulose (DCMC) crosslinked FK films plasticized with glycerol were prepared by a casting method. The effect of DCMC crosslinking on the microstructure, light transmission, aggregate structure, tensile properties, water resistance and water vapor barrier were investigated. The results indicated the formation of both covalent and hydrogen bonding between FK and DCMC to form amorphous FK/DCMC films with good UV-barrier properties and transmittance. However, with increasing DCMC content, a decrease in tensile strength of the FK films indicated that plasticization, induced by hydrophilic properties of the DCMC, partly offset the crosslinking effect. Reduction in the moisture content, solubility and water vapor permeability indicated that DCMC crosslinking slightly reduced the moisture sensitivity of the FK films. Thus, DCMC crosslinking increased the potential viability of the FK films for food packaging applications, offering a value-added product.

Preparation and Characterization of ß-glucosidase Films for Stabilization and Handling in Dry Configurations.

BACKGROUND: Although the stability of proteins is of significance to maintain protein function for therapeutical applications, this remains a challenge. Herein, a general method of preserving protein stability and function was developed using gelatin films. METHODS: Enzymes immobilized onto films composed of gelatin and Ethylene Glycol (EG) were developed to study their ability to stabilize proteins. As a model functional protein, ß-glucosidase was selected. The tensile properties, microstructure, and crystallization behavior of the gelatin films were assessed. RESULTS: Our results indicated that film configurations can preserve the activity of ß-glucosidase under rigorous conditions (75% relative humidity and 37°C for 47 days). In both control films and films containing 1.8 % ß-glucosidase, tensile strength increased with increased EG content, whilst the elongation at break increased initially, then decreased over time. The presence of ß-glucosidase had a negligible influence on tensile strength and elongation at break. Scanning electron-microscopy (SEM) revealed that with increasing EG content or decreasing enzyme concentrations, a denser microstructure was observed. CONCLUSION: In conclusion, the dry film is a promising candidate to maintain protein stabilization and handling. The configuration is convenient and cheap, and thus applicable to protein storage and transportation processes in the future.

Preparation of eicosapentaenoic acid ethyl ester from fish oil ethyl esters by continuous batch chromatography.

Fish oils are rich in eicosapentaenoic acid, which has the wide-ranging biological activities. The rapid and efficient separation of eicosapentaenoic acid ethyl ester from fish oils ethyl ester is still regarded as a challenge. In this study, we described an effective and flexible chromatography for eicosapentaenoic acid ethyl ester preparation, named continuous batch chromatography, which combined the batch chromatography with the continuous chromatographic operation mode. After continuous batch chromatography experiment, the recovery of eicosapentaenoic acid ethyl ester was 82.01%, the average relative purity and the relative highest purity of eicosapentaenoic acid ethyl ester were 97.82 and 98.98%. The productivity of continuous batch chromatography was 5.48 times higher than that of batch chromatography, while the solvent consumption of eicosapentaenoic acid ethyl ester was 78% of the batch chromatography. This study provided a reference for the separation of the targeted chemical component from multi-component mixtures.

The Mitochondrial Endonuclease M20 Participates in the Down-Regulation of Mitochondrial DNA in Pollen Cells.

Maintaining the appropriate number of mitochondrial DNA (mtDNA) molecules is crucial for supporting mitochondrial metabolism and function in both plant and animal cells. For example, a substantial decrease in mtDNA levels occurs as a key part of pollen development. The molecular mechanisms regulating mtDNA copy number are largely unclear, particularly with regard to those that reduce mtDNA levels. Here, we identified and purified a 20-kD endonuclease, M20, from maize (Zea mays) pollen mitochondria. We found M20 to be an His-Asn-His/Asn (H-N-H/N) nuclease that degrades linear and circular DNA in the presence of Mg2+ or Mn2+ Arabidopsis (Arabidopsis thaliana) AtM20, which shared high sequence similarity with maize M20, localized to the mitochondria, had a similar H-N-H/N structure, and degraded both linear and circular DNA. AtM20 transcript levels increased during pollen development, in parallel with a rapid reduction in mtDNA. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 genome-editing techniques were used to generate knockout lines of AtM20 (atm20), which exhibited a significant delay in the reduction in mtDNA levels in pollen vegetative cells but normal mtDNA levels in somatic cells. The delayed reduction in pollen mtDNA levels was rescued by the transgenic expression of AtM20 in atm20 plants. This study thus uncovers an endonucleolytic DNase in plant mitochondria and its crucial role in reducing mtDNA levels, pointing to the complex mechanism regulating mtDNA levels in plants.

A perforated microhole-based microfluidic device for improving sprouting angiogenesis in vitro.

Microfluidic technology is an important research tool for investigating angiogenesis in vitro. Here, we fabricated a polydimethylsiloxane (PDMS) microfluidic device with five cross-shaped chambers using a coverslip molding method. Then, the perforated PDMS microhole arrays prepared by soft lithography were assembled in the device as barriers; a single microhole had a diameter of 100 µm. After injecting type I collagen into the middle gel chamber, we added a culture medium containing a vascular endothelial growth factor (VEGF) into the middle chamber. It would generate a linear concentration gradient of VEGF across the gel region from the middle chamber to the four peripheral chambers. Human umbilical vein endothelial cells (HUVECs) were then seeded on the microhole barrier. With VEGF stimulation, cells migrated along the inner walls of the microholes, formed annularly distributed cell clusters at the gel-barrier interface, and then three-dimensionally (3D) sprouted into the collagen scaffold. After 4 days of culture, we quantitatively analyzed the sprouting morphogenesis. HUVECs cultured on the microhole barrier had longer sprouts than HUVECs cultured without the barrier (controls). Furthermore, the initial distribution of sprouts was more regular and more connections of tube-like structures were generated when the microhole barrier was used. This study introduces a novel microfluidic device containing both microtopographic structures and 3D collagen. HUVECs cultured with the microhole barrier could form well-interconnected tube-like structures and are thus an ideal in vitro angiogenesis model.

Spraying Brassinolide improves Sigma Broad tolerance in foxtail millet (Setaria italica L.) through modulation of antioxidant activity and photosynthetic capacity.

To explore the role of Brassinolide (BR) in improving the tolerance of Sigma Broad in foxtail millet (Setaria italica L.), effects of 0.1 mg/L of BR foliar application 24 h before 3.37 g/ha of Sigma Broad treatment at five-leaf stage of foxtail millet on growth parameters, antioxidant enzymes, malondialdehyde (MDA), chlorophyll, net photosynthetic rate (P N), chlorophyll fluorescence and P700 parameters were studied 7 and 15 d after herbicide treatment, respectively. Results showed that Sigma Broad significantly decreased plant height, activities of superoxide dismutase (SOD), chlorophyll content, P N, PS II effective quantum yield (Y (II)), PS II electron transport rate (ETR (II)), photochemical quantum yield of PSI(Y (I)) and PS I electron transport rate ETR (I), but significantly increased MDA. Compared to herbicide treatment, BR dramatically increased plant height, activities of SOD, Y (II), ETR (II), Y (I) and ETR (I). This study showed BR pretreatment could improve the tolerance of Sigma Broad in foxtail millet through improving the activity of antioxidant enzymes, keeping electron transport smooth, and enhancing actual photochemical efficiency of PS II and PSI.

Investigation of Film with ß-Galactosidase Designed for Stabilization and Handling in Dry Configuration.

Gelatin-based films with an immobilized enzyme designed for extending the stability of the protein in dry, non-powder configuration with precise dosing attributes were subjected to stress conditions of temperature and relative humidity. ß-galactosidase was used as model functional protein. The film configuration preserved the activity of the enzyme under the different storage conditions investigated, which include room temperature under low (ambient) and high (75%) relative humidity, and 36 °C under low (oven) and high relative humidity conditions for a period of 46 days. The influence of the enzyme and plasticizer (glycerol) on the physical and mechanical properties of the films was investigated using DMA (dynamic mechanical analysis). Films containing 5% ß-galactosisdase and glycerol concentrations of 14% or greater exhibited greater tensile strength, Young's modulus, and elongation at break than films with equal concentrations of plasticizer but devoid of any enzyme. The surface texture of the films was analyzed using scanning electron microscopy (SEM). ß-galactosidase and glycerol have opposite effects on the surface morphology of the films. Increasing concentrations of the enzyme result in rougher film surface, whereas increasing the concentration of glycerol leads to films with denser and smoother surface. The results obtained suggest that the dry film configuration approach can help in facilitating the stabilization, handling, storage, and transportation of functional proteins in a cost effective manner.

A microwell pattern for C17.2 cell aggregate formation with concave cylindrical surface induced cell peeling.

We have developed a polydimethylsiloxane (PDMS) pattern with arrays of microwells for the formation of multicellular aggregates by C17.2 neural stem cells. Upon interfacing with the patterns, the neural stem cells would firstly attach to the microwell sidewalls, forming cellular strips on day 1 after plating. For channel connected microwells, cellular strips on the concave semi-cylindrical sidewall surfaces continued among wells and through channels, followed by strip peeling due to prestress arising from actin filaments and assembly of suspending cellular aggregates within the microwells in the following 1-2 days. Our results also suggested that a small microwell diameter of 80 and 100 µm and a narrow channel width of 20 µm would facilitate the aggregate formation among the structural dimensions tested. Finite element method (FEM) simulation revealed that cellular strips on the semi-cylindrical sidewall surfaces peeled under significantly smaller prestresses (critical peeling prestress, CPP), than cells on flat substrates. However, the CPP by itself failed to fully account for the difference in aggregate inducing capability among the patterns addressed, suggesting cell growth behaviors might play a role. This study thus justified the current patterning method as a unique and practical approach for establishing 3D neural stem cell-based assay platforms.

Photosynthetic physiological response of Radix Isatidis (Isatis indigotica Fort.) seedlings to nicosulfuron.

Radix Isatidis (Isatis indigotica Fort.) is one of the most important traditional Chinese medicine plants. However, there is no suitable herbicide used for weed control in Radix Isatidis field during postemergence stage. To explore the safety of sulfonylurea herbicide nicosulfuron on Radix Isatidis (Isatis indigotica Fort.) seedlings and the photosynthetic physiological response of the plant to the herbicide, biological mass, leaf area, photosynthetic pigment content, photosynthetic rate, chlorophyll fluorescence characteristics, and P700 parameters of Radix Isatidis seedlings were analyzed 10 d after nicosulfuron treatment at 5th leaf stage in this greenhouse research. The results showed that biological mass, total chlorophyll, chlorophyll a, and carotenoids content, photosynthetic rate, stomatal conductance, PS II maximum quantum yield, PS II effective quantum yield, PS II electron transport rate, photochemical quenching, maximal P700 change, photochemical quantum yield of PS I, and PS I electron transport rate decreased with increasing herbicide concentrations, whereas initial fluorescence, quantum yield of non-regulated energy dissipation in PS II and quantum yield of non-photochemical energy dissipation due to acceptor side limitation in PS I increased. It suggests that nicosulfuron ≥1 mg L-1 causes the damage of chloroplast, PS II and PS I structure. Electron transport limitations in PS I receptor side, and blocked dark reaction process may be the main cause of the significantly inhibited growth and decreased photosynthetic rate of Radix Isatidis seedlings.

Responsiveness of voltage-gated calcium channels in SH-SY5Y human neuroblastoma cells on quasi-three-dimensional micropatterns formed with poly (l-lactic acid).

INTRODUCTION: In this study, quasi-three-dimensional (3D) microwell patterns were fabricated with poly (l-lactic acid) for the development of cell-based assays, targeting voltage-gated calcium channels (VGCCs). METHODS AND MATERIALS: SH-SY5Y human neuroblastoma cells were interfaced with the microwell patterns and found to grow as two dimensional (2D), 3D, and near two dimensional (N2D), categorized on the basis of the cells' location in the pattern. The capability of the microwell patterns to support 3D cell growth was evaluated in terms of the percentage of the cells in each growth category. Cell spreading was analyzed in terms of projection areas under light microscopy. SH-SY5Y cells' VGCC responsiveness was evaluated with confocal microscopy and a calcium fluorescent indicator, Calcium Green™-1. The expression of L-type calcium channels was evaluated using immunofluorescence staining with DM-BODIPY. RESULTS: It was found that cells within the microwells, either N2D or 3D, showed more rounded shapes and less projection areas than 2D cells on flat poly (l-lactic acid) substrates. Also, cells in microwells showed a significantly lower VGCC responsiveness than cells on flat substrates, in terms of both response magnitudes and percentages of responsive cells, upon depolarization with 50 mM K(+). This lower VGCC responsiveness could not be explained by the difference in L-type calcium channel expression. For the two patterns addressed in this study, N2D cells consistently exhibited an intermediate value of either projection areas or VGCC responsiveness between those for 2D and 3D cells, suggesting a correlative relation between cell morphology and VGCC responsiveness. CONCLUSION: These results suggest that the pattern structure and therefore the cell growth characteristics were critical factors in determining cell VGCC responsiveness and thus provide an approach for engineering cell functionality in cell-based assay systems and tissue engineering scaffolds.