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The environmental gas concentration affects the storage period and quality of fruits and vegetables. High concentration CO₂ treating for a long time will cause damage to fruits, However, the specific molecular mechanism is unclear. To analyze the mechanism of CO₂ injury in apple, high-throughput sequencing technology of Illumina Hiseq 4000 and non-targeted metabolism technology were used to analyze the transcriptome sequencing and metabolomics analysis of browning flesh tissue of damage fruit and normal pulp tissue of the control group. A total of 6 332 differentially expressed genes were obtained, including 4 187 up-regulated genes and 2 145 down regulated genes. Functional analysis of the differentially expressed genes confirmed that the occurrence of CO₂ injury in apple was related to redox process, lipid metabolism, hormone signal transduction process and energy metabolism process. Twenty candidate browning genes were successfully screened, among which grxcr1 (md14g1137800) and gpx (md06g1081300) participated in the reactive oxygen species scavenging process, and pld1_ 2 (md15g1125000) and plcd (md07g1221900) participated in phospholipid acid synthesis and affected membrane metabolism. mdh1 (md05g1238800) participated in TCA cycle and affected energy metabolism. A total of 77 differential metabolites were obtained by metabolomic analysis, mainly organic acids, lipids, sugars and polyketones, including 35 metabolites related to browning. The metabolism of flavonoids was involved in the browning process of apple. Compared with the control tissue, the content of flavonoids such as catechin and quercetin decreased significantly in the damaged apple tissue, the antioxidant capacity of cells decreased, the redox state was unbalanced, and the cell structure was destroyed, resulting in browning. The results of this study further enrich the theoretical basis of CO₂ damage, and provide reference for the practical application of high concentration CO₂ preservation technology.

Fruit cracking is a common physiological disease. Many fruits such as tomato, sweet cherry, apple, jujube, pomegranate, and litchi are liable to crack, causing considerable economic loss and agricultural resources waste. The mechanisms of fruit cracking are comprehensive. Some correlations have been observed between susceptibility of fruit cracking and some fruit traits (genetic, fruit size, fruit shape, fruit growth rate, water content, fruit skin characteristics, related gene expression, etc). Also, environmental condition (temperature, light, rainfall, etc) and orchard management (irrigation, sun-shade, mineral, growth regulator, etc) can influence fruit cracking. Here, progress in studies on fruit cracking is reviewed to provide a reference for prevention and control of fruit cracking.

Table grape (Vitis vinifera L.) is one of the most popular berries in the world but easy to decay due to gray mold infiltration during storage. Sulfur dioxide (SO2) treatment is by far the most efficient way of for controlling gray mold development and long-term preserving table grape fruit. However, SO2 treatment causes severe abscission of berry from fruit cluster. In this study, two edible coatings (1.5% chitosan and 1.0% poly-e- lysine) were selected to prevent the berry from SO2-induced abscission in ‘Kyoho’ table grapes (Vitis vinifera L. x V. Labrusca) during room temperature (20 ℃) storage. Results indicated that both 1.5% chitosan and 1.0% poly-e- lysine treatments remarkably inhibited the SO2-induced abscission in ‘Kyoho’ fruits, and the abscission rates of both coating treated-groups were significantly lower even than the blank control (without SO2 treatment) after 6 d of storage. Meanwhile, both edible coating treatments significantly decreased fruit weight loss and inhibited the increase of the cell-wall-degrading enzyme activities. Overall, these two edible coating treatments maintained other fruit qualities such as, total soluble solids (TSS), titratable acids (TA), and content of vitamin C. Thus, the edible coating 1.5% chitosan or 1.0% poly-e- lysine provide potential effective ways to control SO2-induced fruit abscission and quality deterioration in table grape, and they may be of great commercial value during postharvest handling of table grape.

The small chemical compound 8-ethoxy-2-(4-fluorophenyl)-3-nitro-2H-chromene (S14161) was recently identified as an inhibitor of phosphoinositide 3-kinase (PI3K) and reported to inhibit tumor growth. However, its chiral structure and poor solubility prevent its further use. Compound 6-bromo-8-ethoxy-3-nitro-2H-chromene (BENC-511) is an analogue of S14161 produced by structural optimization. A previous study indicated that BENC-511 acted on multiple myeloma and that it had a toxicity by inhibiting the PI3K/protein kinase B (Akt) pathway. However, the effects of BENC-511 on the proliferation and apoptosis of A549 human lung adenocarcinoma cells have not been reported. The current study investigated the effects of BENC-511 on the proliferation and apoptosis of A549 cells in vitro. Results indicated that the compound BENC-511 inhibited the viability of A549 cells in a concentration- and time- dependent manner. BENC-511 suppressed proliferation and colony formation via S phase arrest. BENC-511 decreased the expression of cyclin A, proliferating cell nuclear antigen (PCNA), B-cell lymphoma-2 (Bcl-2), phospho-mammalian target of rapamycin (p-mTOR), and phospho-Akt (p-Akt) and it increased the expression of p21WAF1CIP1(p21), Caspase-3 and Caspase-9. In conclusion, BENC-511 inhibited the proliferation of A549 human lung adenocarcinoma cells via S phase arrest as a result of up-regulation of p21 and reduction of Cyclin A/cyclin-dependent kinase 2 (CDK2)/PCNA complex and it induced apoptosis by reducing the mitochondrial membrane potential via the Akt/Bcl-2/Caspase-9 mitochondrial pathway of apoptosis.

Edwardsiella tarda is one of the leading fish pathogens for the aquaculture industry. To realize efficient disease control of edwardsiellosis, a predictive model for E. tarda in seawater was developed. The modified logistic model was used to regress the growth curves of E. tarda JN at five different temperatures (range from 10 to 30 °C) and four organic nutrient concentrations (range from 5 to 40 mg l−1 measured by chemical oxygen demand (COD)). The modeling effects of temperature and COD on the specific growth rate (μ) were developed by square-root model and saturation-growth rate model, respectively. The growth model was validated in turbot aquaculture tanks by estimating the dynamics of inoculated E. tarda. The accurate feeding of probiotic Bacillus pumilus strain H2 was calculated based on the estimation of E. tarda. Results showed that the logistic model produced a good fit to the growth curves of E. tarda JN (average R2 = 0.962). The overall predictions based on above models agreed well with the growth curve of E. tarda JN observed by plate counting in the validation tests (average Af = 1.16; average Bf = 1.32). The use of predicted amount of B. pumilus (5.66 log CFU ml−1) successfully prevent the deterioration of disease for turbot with 13.3% mortality rate in a recirculating aquaculture system (RAS), while the feeding of 0 and 3.0 log CFU ml−1 of B. pumilus resulted in 53.7 and 75.3% of turbot mortality rate, respectively. In conclusion, accurate estimation of E. tarda realized the precise feeding of probiotics, which successfully prevent the rapid progression of the edwardsiellosis.

Acute hepatopancreatic necrosis disease (AHPND) outbreaks in cultured shrimps were identified in Zhangpu, China. One Vibrio parahaemolyticus strain PB1937 was isolated from the cultured shrimps and was confirmed as a causative agent of the AHPND outbreak by employing Koch’s four postulates. Challenge tests with 106 cells ml−1 of strain PB1937 caused 100% mortality of shrimps, indicating it had sufficient virulence to cause the outbreak. Phylogenomic analysis revealed a clear divergence between PB1937 and 14 publicly available V. parahaemolyticus strains and divided 11 AHPND-causing strains into six genomic clusters. Prophage profiling of above strains revealed strong correlations with their genomic relationship, while Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) were almost absent in the genomes. The binary toxin gene pirABvp directly related to the development of AHPND was found in a 70-kb plasmid p1937-1 in PB1937 but was absent in a 78-kb novel plasmid p1937-2, which shared 46% sequence similarity with p1937-1. Comparative genomic analysis revealed that PB1937 has a novel truncated type VI secretion system (T6SS1) which possibly affects its antibacterial activity. In addition, three novel genomic islands were reported. The analysis of the genomes gave some clues regarding the correlation of virulence with its genomic trait for the AHPND strains.

Fish challenged with pathogens often show behavioral disorders. The present study investigated the influence of Aeromonas salmonicida infection on the body color changes of Atlantic salmon (Salmo salar L.). In both conditions of a white background and transfer from the black background to the white background, the body color contrast of the fish infected with A. salmonicida (100 μL bacterium solution 8.2 × 109 CFU ml−1 in the dorsal muscle) declined significantly following an initial increase compared with the control group (100 μL 0.9 % NaCl); the greatest differences were observed on day 11, at 57.6 % and 52.7 % respectively (P

SO2 treatment is currently the preferred method for postharvest preservation of table grapes, while inappropriate amount of environmental SO2 would render kinds of physiological damage, including tissues bleaching, flavor destroying and berry abscission. Our previous study has reported that 1.5% chitosan solution treatment could inhibit postharvest berry abscission of the ‘Kyoho’ table grapes, especially caused by excessive SO2 treatment. However, the physiological mechanism of chitosan inhibiting SO2-induced berry abscission still remains unclear. This study further focused on physiological changes of the abscission zone (AZ) between stalk and pedicle, including fruit detachment force (FDF), the microscopic structure, cell wall degrading enzymes and SO2 permeation, during storage of the ‘Kyoho’ table grapes with different postharvest treatments. The results showed that the 1.5% chitosan treatment remarkably suppressed the decrease of FDF, and delayed the structural deterioration in AZ during storage. Meanwhile, this chitosan treatment also distinctly suppressed the increase of cell wall degrading enzymes activity and SO2 permeation into AZ, which showed statistically linear relationship with abscission index. Those results contribute to elucidate the physiological mechanism of chitosan-inhibited berry abscission and develop new preservation technology with greater commercial value for postharvest table grapes in the future.

In this research, astaxanthin-rich nanopowder was prepared by nanoencapsulation and freeze-drying techniques with enhanced bioavailability and antioxidant activities. The nanopowder showed a maximum solubility of 230 mg mL−1 with an astaxanthin content as high as 2.9%. Compared with free astaxanthin, the astaxanthin-loaded nanopowder exhibited a more efficient antioxidant effect: an oral dose of 0.9 mg per kg BW significantly reduced the malondialdehyde and protein carbonyl contents, and increased the glutathione content as well as the superoxide dismutase activities in alcohol-induced acute hepatic injured mice, and maintained these oxidative stress indicators at a normal level for a longer period when treated with nanoencapsulated-astaxanthin than free astaxanthin. Simulated gastrointestinal tract studies demonstrated that the nanopowder with pH and DNase I-dependent dissociation properties delivered astaxanthin efficiently to the small intestine. Astaxanthin-rich nanopowder with a dose as high as 2.4 mg per kg BW (equivalent to astaxanthin) showed no chronic toxicity to mice in terms of hematology and pathological histology, indicating its impressive biocompatibility for biomedical applications. Pharmacokinetics and relative bioavailability (207%) of the nanopowder further proved that DNA/chitosan nanocarriers significantly improved the delivery efficiency of astaxanthin. With enhanced bioavailability and antioxidant activities, this novel type of astaxanthin-loaded nanopowder is expected to find broad application in the food and drug industry.

Diabetes mellitus-related cardiomyopathy (DMCMP) has been defined as ventricular dysfunction that occurs in diabetic patients independent of a recognized cause such as coronary artery disease or hypertension. Mechanisms underlying DMCMP have not been fully elucidated. In this study, the roles of protein phosphatase 2A/nuclear factor NF-E2-related factor 2 (PP2A/Nrf2) in experimental DMCMP induced by high glucose were studied in vitro and in vivo. The results showed that high glucose could induce experimental DMCMP and increase ROS generation, increase the expression and nuclear translocation of Nrf2, down-regulate the expression of PI3K/Akt/mTOR and up-regulate the expression of ERK, and activate the autophagy of cardiomyocytes. The activity or expression of PP2A in DMCMP increased. PP2A could up-regulate the expression of Nrf2 and promote cardiomyocytes autophagy and apoptosis. Inhibition of PP2A could reduce the expression of Nrf2 and inhibit the autophagy and apoptosis of cardiomyocytes. The results suggested that hyperglycemic-induced experimental DMCMP may be related to up-regulating the expression of Nrf2 through PP2A/Nrf2 pathway. These results will be helpful to elucidate the pathogenesis and mechanism of DMCMP and find targets for the development of new drugs to prevent or treat DMCMP.

Although it has been hypothesized that a predictable feeding regime in animals allows physiological variables to be adjusted to maximize nutrient utilization and, hence, better growth performance, the assumption has rarely been tested. This study compares the effects of time-restricted versus free access self-feeding on the growth, feeding behavior and daily digestive enzyme rhythms of Atlantic salmon (Salmo salar). In an experiment that lasted 6 weeks, fish (109.9 g) were divided into two groups: group 1 had free access to a self-feeder (FA); group 2 received three meals per day (2 h per meal) at dawn, midday and dusk via a time-restricted self-feeder (TR). At the end of the experiment, the fish were sampled every 3 h over a 24-h period. The results showed that the TR fish quickly synchronized their feeding behavior to the feeding window and their blood glucose showed a significant postprandial increase, while FA fish displayed no statistically significant rhythms (P>0.05). Pepsin activity of TR fish also showed a significant daily rhythm (P 0.05). In conclusion, the study failed to confirm a link between the entrainment of daily digestive enzyme profiles and growth performance, with the TR group showing comparatively poor blood glucose regulation.

Despite many studies about functions of miR396 were concentrated on cotyledon and leaf growth and development, only few researches were focused on flower and fruit, especially for fleshy fruit, for example, tomato fruit. Here, the roles of miR396 throughout the growth and development of tomato plant were explored with combining bioinformatics and transgene-mediated methods. In tomato, miR396 had two mature types (miR396a and miR396b), and miR396a expressed significantly higher than miR396b in cotyledon, flower, sepal and fruit. Generally, plant growth and development were regulated by miR396 via growth-regulating factors (GRFs). In tomato, all 13 SlGRFs were analyzed comprehensively, including phylogeny, domain and expression patterns. To investigate the roles of miR396 further, STTM396a/396a-88 was over-expressed in tomato, which induced miR396a and miR396b both dramatical down-regulation, and the target GRFs general up-regulation. As a result, the flowers, sepals and fruits all obviously became bigger. Most significantly, the sepal length of transgenic lines #3 and #4 at 39 days post-anthesis was separately increased 75% and 81%, and the fruit weight was added 45% and 39%, respectively. Overall, these results revealed novel roles of miR396 in regulating flower and fruit development, and provided a new potential way for improving tomato fruit yield.

The way in which the feed is delivered mainly influences the growth rates and feed efficiency. Restricting the time during which food is available from a self-feeder may help to regulate production parameters, but may also increase competition. In this article, the effects of time-restricted self-feeding on growth, feeding behavior and fin index Atlantic salmon (Salmo salar) were studied over the course of 6 weeks. A total of 150 fish were divided into two groups in triplicates: One group had free access to self-feeders (FA), and the other group received three meals (2 h per meal) restricted to dawn, midday and dusk a day in light phase (TR). The results showed that there were no significant differences in production parameters (specific growth rate 1.33 ± 0.21 and 1.40 ± 0.21 % day−1, for TR and FA, respectively), body composition and nutrient retention efficiencies between treatments, while the FA fish had significantly higher food wastage (0.79 ± 0.06 vs. 0.56 ± 0.10 %) compared with the TR fish. In the course of the experiment, the TR fish gradually improved the accuracy of trigger actuation; in 1–2, 3–4 and 5–6 weeks, 61, 68 and 80 % trigger actuations happened in the feeding window. Nevertheless, there was a significant decrease in the dorsal fin index found in the TR fish as the experiment proceeded (9.83 ± 0.15 and 8.56 ± 0.04 for the beginning and end, respectively; P

Numerous studies have been focusing on breeding tomato plants with enhanced lycopene accumulation, considering its positive effects of fruits on the visual and functional properties. In this study, we used a bidirectional strategy: promoting the biosynthesis of lycopene, while inhibiting the conversion from lycopene to β- and α-carotene. The accumulation of lycopene was promoted by knocking down some genes associated with the carotenoid metabolic pathway. Finally, five genes were selected to be edited in genome by CRISPR/Cas9 system using Agrobacterium tumefaciens-mediated transformation. Our findings indicated that CRISPR/Cas9 is a site-specific genome editing technology that allows highly efficient target mutagenesis in multiple genes of interest. Surprisingly, the lycopene content in tomato fruit subjected to genome editing was successfully increased to about 5.1-fold. The homozygous mutations were stably transmitted to subsequent generations. Taken together, our results suggest that CRISPR/Cas9 system can be used for significantly improving lycopene content in tomato fruit with advantages such as high efficiency, rare off-target mutations, and stable heredity.

RNA editing plays a key posttranscriptional role in gene expression. Existing studies on cytidine-to-uridine RNA editing in plants have focused on maize (Zea mays), rice (Oryza sativa), and Arabidopsis (Arabidopsis thaliana). However, the importance and regulation of RNA editing in several critical agronomic processes are not well understood, a notable example of which is fruit ripening. Here, we analyzed the expression profile of 33 RNA editing factors and identified 11 putative tomato (Solanum lycopersicum) fruit ripening-related factors. A rapid virus-induced gene silencing assay indicated that the organelle RNA recognition motif-containing protein SlORRM4 affected tomato fruit ripening. Knocking out SlORRM4 expression using a clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 genome editing strategy delayed tomato fruit ripening by lowering respiratory rate and ethylene production. Additionally, the expression of numerous genes associated with fruit ripening and mitochondrial functions changed significantly when SlORRM4 was knocked out. Moreover, the loss of SlORRM4 function significantly reduced RNA editing of many mitochondrial transcripts, leading to low-level expression of some core subunits that are critical for mitochondrial complex assembly (i.e. Nad3, Cytc1, and COX II). Taken together, these results indicate that SlORRM4 is involved in RNA editing of transcripts in ripening fruit that influence mitochondrial function and key aspects of fruit ripening.

Summary In recent years, the type II CRISPR system has become a widely used and robust technique to implement site‐directed mutagenesis in a variety of species including model and crop plants. However, few studies manipulated metabolic pathways in plants using the CRISPR system. Here, we introduced the pYLCRISPR/Cas9 system with one or two single‐site guide RNAs to target the tomato phytoene desaturase gene. An obvious albino phenotype was observed in T0 regenerated plants, and more than 61% of the desired target sites were edited. Furthermore, we manipulated the γ‐aminobutyric acid (GABA) shunt in tomatoes using a multiplex pYLCRISPR/Cas9 system that targeted five key genes. Fifty‐three genome‐edited plants were obtained following single plant transformation, and these samples represented single to quadruple mutants. The GABA accumulation in both the leaves and fruits of genomically edited lines was significantly enhanced, and the GABA content in the leaves of quadruple mutants was 19‐fold higher than that in wild‐type plants. Our data demonstrate that the multiplex CRISPR/Cas9 system can be exploited to precisely edit tomato genomic sequences and effectively create multisite knockout mutations, which could shed new light on plant metabolic engineering regulations.

Predicting the growth of key microorganisms is essential to improve the efficiency of wastewater treatment of recirculating aquaculture systems (RAS). We have developed a stochastic model to assess quantitatively the microbial populations in RAS. This stochastic model encompassed the growth model into the Monte Carlo simulation and was constructed with risk analysis software. A modified logistic model combined with the saturation growth-rate model was successfully developed to regress the growth curves of six microorganisms. Monte Carlo simulation was employed to model the effects of chemical oxygen demand (COD) on the maximum specific growth rate. Probabilistic distributions and predictions under the different COD ranges were generated for each simulated scenario. The coefficient of determination (R 2) and bias factor (Bf) were used to assess the performance of an established model. Logistic model produced a good fit to the growth curve of Flavobacterium sp. (R 2 = 0.9511), Acinetobacter baumannii (R 2 = 0.9970), Sphingomonas paucimobilis (R 2 = 0.9086), Vibrio natriegens (R 2 = 0.9993), Lutimonas sp. (R 2 = 0.9872) and Bacillus pumilus (R 2 = 0.9816). Bacterial population structure was determined by the construction of 16S rRNA gene libraries. A regular variation trend was observed for the dominant groups during the entire process, with a decrease of Cytophaga–Flavobacterium–Bacteroidetes from 37.6 to 18.7 % and an increase in Gammaproteobacteria from 8.5 to 30.6 %. The predicted model agreed well with observed values except for Flavobacterium sp., and the results can be applied to predict key microorganisms in actual environments. The results of this study provide a method to monitor the dynamics of key microorganisms, which can also help to evaluate the impacts of microorganisms on the operations of RAS.

Green-ripe (Gr) tomato carries a dominant mutation and yields a nonripening fruit phenotype. The mutation results from a 334 bp deletion in a gene of unknown function at the Gr locus. The putative influence of Gr gene-deletion mutation on biochemical changes underlying the nonripening phenotype remains largely unknown. Respiration of Gr fruit was found to be reduced at mature green and breaker stage of ripening, while the fruit softening was dramatically prolonged. We studied the proteome of Gr mutant fruit using high-throughput iTRAQ and high-resolution mass spectrometry and identified 43 proteins representing 43 individual genes as potential influence-targets of Gr mutated fruit. The identified proteins are involved in several ripening-related pathways including cell-wall metabolism, photosynthesis, oxidative phosphorylation, carbohydrate and fatty acid metabolism, protein synthesis, and processing. Affected protein levels are correlated with the corresponding gene transcript levels. The modulation in the accumulation levels of PI(U1)P, PGIP, and PG2 supported the delayed softening phenotype of the Gr fruit. Further investigation in GR gene-silencing fruit ascertained the doubtless modulation of these targets by the deletion mutation of GR gene.

Genotype is generally determined by the co-expression of diverse genes and multiple regulatory pathways in plants. Gene co-expression analysis combining with physiological trait data provides very important information about the gene function and regulatory mechanism. L-Ascorbic acid (AsA), which is an essential nutrient component for human health and plant metabolism, plays key roles in diverse biological processes such as cell cycle, cell expansion, stress resistance, hormone synthesis, and signaling. Here, we applied a weighted gene correlation network analysis approach based on gene expression values and AsA content data in ripening tomato (Solanum lycopersicum L.) fruit with different AsA content levels, which leads to identification of AsA relevant modules and vital genes in AsA regulatory pathways. Twenty-four modules were compartmentalized according to gene expression profiling. Among these modules, one negatively related module containing genes involved in redox processes and one positively related module enriched with genes involved in AsA biosynthetic and recycling pathways were further analyzed. The present work herein indicates that redox pathways as well as hormone-signal pathways are closely correlated with AsA accumulation in ripening tomato fruit, and allowed us to prioritize candidate genes for follow-up studies to dissect this interplay at the biochemical and molecular level.

The background colour of aquaculture tanks is normally chosen based on practical experience and/or observations of fish behaviour and the growth rates achieved. However, some farmed species, including turbot, are sentient and can show a preference for a particular environment. In the current study, a self-referent colour preference device was developed and the self-referent colour preference of farmed fish investigated. In experiment 1, the background colour preference of juvenile turbot cultured under a grey background for >3 months post-incubation was evaluated. Based on these results, in experiment 2, juvenile turbot were adapted to blue, pink, white, or black backgrounds for 50 days and their preferences established. Meanwhile, the growth rates, feed intake, and metabolic rates (including oxygen consumption rate, and ammonia excretion rate) of the turbot were evaluated. The results showed that turbot farmed under a grey background, or after long-term white, blue, pink and black colour adaptation, always displayed a preference for a white background and a dislike for black, red, or brown backgrounds, although their body colour was greyish. Long-term adaptation influenced the frequency of juveniles selecting white, black, pink or blue backgrounds. They showed the highest growth rate, feed intake, and metabolic rates under blue and white backgrounds, and the lowest under a black background in accordance with their preferences shown in experiment 1. Although it is unclear how turbot determine their self-referent colour preferences over such a short period of time, these results indicate that dark colours are unsuitable for the aquaculture of turbot culture in terms of the welfare of the fish. (C) 2016 Elsevier Inc. All rights reserved.

The plasmid pGSA1285 was first modified by substituting its GUS sequence with the Chalcone synthase intron fragment from vector pFGC5941 to get the plant silencing expression vector that contained Kanamycin resistance site and was named as pGSA2285. Using PCR-based amplification, two different restriction sites at both ends of tobacco Bax inhibitor-1 (NtBI-1) gene were created, respectively, which made the construction of ihpRNA gene silencing vector more efficiently. Then, NtBI-1 genes were inserted into Multiple Cloning Site (MCS) of pGSA2285 respectively to form Bax inhibitor-1 ihpRNA gene silencing vector, named as pGSA4285, containing sense and anti-sense BI-1 sequence which was spliced by chalcone synthase intron. Combined PCR identification and enzyme restriction analyses, the results showed that Bax inhibitor-1 ihpRNA gene silencing vector had been constructed and transferred into Agrobacterium tumefaciens EHA105 successfully, which laid a foundation for the further study on the function of BI-1 in plant PCD regulation.

Plums easily show internal browning (IB) after low temperature storage. Polyphenol oxidase (PPO) was recognized as the main reason for IB of many kinds of fruits. The aim of this study was to develop a vacuum infiltration-based method with ascorbic acid (AsA) to inhibit plum PPO. The results showed that browning rate and PPO activity of plum fruits (Prunus salicina Lindell cv. Yuhuang) were significantly inhibited by vacuum infiltration with 0.5% AsA during 98 days of storage at 3 ± 1C, and its preservation effects, which included inhibition of browning and keeping fruit quality, were better than those produced by vacuum infiltration with 0.1% AsA or dipping treatment with 0.5% AsA. Based on the results, the method of browning inhibition for postharvest plum consisted of 10 min infiltration time, 200 mmHg and 0.5% AsA. PRACTICAL APPLICATIONS Postharvest plum fruits suffer chilling injury phenomenon followed by internal browning that reduces the consumer acceptance and commercial value. However, traditional preservation methods, such as dipping with antistaling agent, are limited in browning inhibition. Thus, a new and efficient method for intact plums stored at 3 ± 1C was developed to reduce the internal browning of postharvest plum fruits, increase the supply of plum fruits with high quality, and enhance the income of farmers and dealers. The application parameters of this vacuum infiltration-based method are based on: 10 min of infiltration time, a vacuity of 200 mmHg, and a concentration of 0.5% of AsA solution.

MicroRNAs (miRNAs) are a newly discovered class of nonprotein-coding small RNAs with the length of ∼21 nucleotides that regulate gene post-transcriptional expression in animals and plants. By far, the researches have indicated that miRNAs may play multiple roles in plant growth and development. It is difficult to identify some miRNAs by experimental methods because of their low expressional levels and tissue specificity, while bioinformatics is an effective strategy in the prediction of this kind of miRNAs. In this study, we presented an approach of expressed sequence tag (EST) analysis for predicting novel miRNAs as well as their targets in tomato (Lycopersicon esculentum). The database of tomato ESTs was compared with previously known miRNA sequences of other plants using BLAST to search for potential miRNAs. Eight potential miRNAs were found following a range of filtering criteria, including stem-loop structure, mismatches, the content of A + U, minimal folding free energy indices, and others with subsequent validated by touchdown RT-PCR assay in fruit tissue. Three unknown miRNAs, LemiR157a, LemiR172i, and LemiR399, which were not reported in previous study, were found in tomato. Tomato mRNA database was further compared with the newly identified miRNA sequences with BLAST, and 42 potential targets of miRNAs were identified. According to the annotations of tomato mRNAs provided by the website (http://ted.bti.cornell.edu/digital/sRNA), miRNA target genes were classified into four groups, in which transcription factors regulating growth and development, signal pathway transduction, and metabolism of tomato plants were in the majority.

As a method of post-transcriptional gene silencing (PTGS), virus-induced gene silencing (VIGS) became a powerful tool for the study of gene function in plants. Current researches are focused on unraveling the mechanisms of PTGS, finding novel viral vectors and new infection methods in order to broaden the host species spectrum, and establishing the new reporter gene system of VIGS to demonstrate gene silencing, as well as developing the high-throughput silencing for genomic scale research. The aim of this review is to bring together recent advances of VIGS, an attractive method, for understanding it in the study of plants.

Luteolin-7-O-glucoside (LUTG) was isolated from the plants of Dracocephalum tanguticum Maxim. Previous research has showed that LUTG pretreatment had a significant protective effect against doxorubicin (DOX)-induced cardiotoxicity by reducing intracellular calcium overload and leakage of creatine kinase and lactate dehydrogenase. But the underlying mechanisms have not been completely elucidated. In the present study, we investigated the effects of LUTG on H9c2 cell morphology, viability, apoptosis, reactive oxygen species generation, and the mitochondrial transmembrane potentials. The expression of p-PTEN, p-Akt, p-ERK, p-mTOR, and p-GSK-3β were detected by Western blotting. Compared with DOX alone treatment group, the morphological injury and apoptosis of the cells in groups treated by DOX plus LUTG were alleviated, cell viability was increased, ROS generation was lowered remarkably, and mitochondrial depolarization was mitigated. In DOX group, the expression of p-PTEN was lower than normal group and the expression of p-Akt and p-ERK was higher than normal group. In the groups treated with LUTG (20 μM), the expression of p-PTEN was upregulated and the expression of p-Akt, p-ERK, p-mTOR, and p-GSK-3β was downregulated. These results indicated that the protective effects of LUTG against DOX-induced cardiotoxicity may be related to anti-apoptosis through PTEN/Akt and ERK pathway.

Background: A variety of processes and etiologies are thought to be involved in the pathophysiology of auditory neuropathy (AN). However, little is known about the clinical and molecular characteristics of hereditary AN. Objective: To explore the clinical and genetic findings of a Chinese family with AN. Methods: Seven patientsin three consecutive generations of the pedigree were selected. Detailed history collection, physical examination, and audiological evaluations including pure-tone audiometry, acoustic immittance, auditory brainstem responses, cochlear microphonics, and evoked otoacoustic emissions, and mitochondrial DNA analysis were performed. Results: All subjects involved are offspring of a female ancestor in the pedigree. In 6 of them, the hearing impairment started before the age of 9. Audiograms showed bilateral, symmetric, and profound deafness. Other audiological examinations revealed absent acoustic reflexes and auditory brainstem responses, and preserved evoked otoacoustic emissions and cochlear microphonics. One subject was characterized by normal audiological findings except high-frequency hearing loss with later onset. Hearing deterioration was found in 2 subjects who were followed for 26 months. Physical examination and mitochondrial DNA analysis yielded normal results. Conclusions: Clinical features in the pedigree are consistent with type II AN. Pedigree analysis and molecular findings indicate an autosomal dominant inheritance.

Inflammatory pseudotumor (IP) is a clinically aggressive but histologically benign condition of unknown cause. Its appearance in the temporal bone is uncommon. We present clinical, radiological, and histopathologic findings of three cases originating in the temporal bone. In the first case, a simultaneous IP of the temporal bone and parotid gland was found with histopathologic confirmation. In the second case, an enlarged cervical node, which was also believed to be related to IP, was observed accompanied with the temporal lesion. While the third case presented with chronic suppurative otitis media. Two of them were treated by surgery alone with complete resolve of the diseases. Another one underwent tympanomastoidectomy in combination with oral steroids, radiation, and chemotherapy, but the IP still recurred. A comprehensive review of the literature on clinical features of the temporal pseudotumor was conducted.

Small RNAs have emerged as critical regulators in the expression and function of eukaryotic genomes at the post-transcriptional level. To elucidate the functions of microRNA (miRNAs) and endogenous small-interfering RNAs (siRNAs) in tomato fruit ripening process, the deep sequencing and bioinformatics methods were combined to parse the small RNAs landscape in three fruit-ripening stages (mature green, breaker and red-ripe) on a whole genome. Two species-specific miRNAs and two members of TAS3 family were identified, 590 putative phased small RNAs and 125 cis-natural antisense (nat-siRNAs) were also found in our results which enriched the tomato small RNAs repository and all of them showed differential expression patterns during fruit ripening. A large amount of the targets of the small RNAs were predicted to be involved in fruit ripening and ethylene pathway. Furthermore, the promoters of the conserved and novel miRNAs were found to contain the conserved motifs of TATA-box and CT microsatellites which were also found in Arabidopsis and rice, and several species-specific motifs were found in parallel.

Elevated levels of hypoxia-inducible factor 1α (HIF-1α) in middle ear effusion may play an important role in the pathogenesis of bone conduction impairment associated with otitis media with effusion (OME). The mechanism may be related to the up-regulation of nitric oxide (NO) expression.This study was undertaken to investigate the role of HIF-1α in the pathogenesis of sensorineural hearing loss associated with OME.One hundred and eight OME patients were divided into two groups: OME without bone conduction impairment (group 1) and OME with bone conduction impairment (group 2). The levels of HIF-1α, NO, and quinolinic acid (QUIN) in the middle ear effusion and serum of these patients were investigated. The relationship between these factors and the bone conduction threshold (BCT) differences were analyzed.The levels of HIF-1α and NO concentrations in the middle ear effusion were found to be significantly higher in group 2 than in group 1 (both p0.05). The OME patients' BCT differences at 4000 Hz were correlated with the levels of HIF-1α and the NO concentrations in the middle ear effusion. Furthermore, the HIF-1α levels were correlated with the levels of NO but not with the levels of QUIN in the effusion.

Background We report an extremely rare case of hypopharyngeal paraganglioma that was accompanied by a paraneoplastic neurologic syndrome (PNS). Methods The clinical, radiological, and histopathologic findings of the patient are presented. Results A 49-year-old woman presented with a history of progressive asthenia of the lower extremities persisting for 2 months. She also experienced numbness and sensory disturbance of the left lower limbs, 2 epileptic seizures, and constipation during the period. Clinical and radiological evaluations suggested the diagnosis of encephalomyelitis in combination with a left hypopharyngeal mass lesion. The patient underwent surgical excision of the lesion, which subsequently resolved her symptoms. Postoperative pathology was consistent with a paraganglioma. There was no evidence of local recurrence or distant metastasis of the tumor on recent follow-up. Conclusions To the best of our knowledge, our patient represents the third reported case of hypopharyngeal paraganglioma, and the only one to have a paraneoplastic syndrome. © 2012 Wiley Periodicals, Inc. Head Neck 35: E304–E307, 2013

To develop and evaluate the improved Chinese hearing questionnaire for school children (CHQS) for mass epidemiology study on hearing impairment in China.Using the probability proportion to size (PPS) method, 8412 residents were investigated in 40 clusters in Jiangsu province with the WHO ear diseases and hearing disorders survey protocol. 87.9% of the residents aged 7 years and over answered the questionnaire and accepted the pure tone audiometry.The prevalence of hearing impairment was 12.9% by the questionnaire. Compared with "golden standard" (pure tone audiometry), Sen = 58.5%, Spe = 96.7%, PV+ = 78.9%, PV- = 91.7%, overall accuracy = 90.0%. The sensitivity for women was higher than men.The questionnaire produced high efficiency and specificity values. It could be used in mass hearing screening, particularly in remote and rural area, although the sensitivity was as low as most questionnaires.

To investigate if the OTOF gene contributes to the non-syndromic hearing loss of a Chinese pedigree with dominantly inherited auditory neuropathy (AN).The subjects included were 9 live individuals in an autosomal dominant AN pedigree, 3 sporadic AN patients and 3 normal-hearing controls. Genomic DNA was isolated from the peripheral leukocytes of the subjects using the Pure gene DNA Isolation Kits. Firstly, the whole coding sequence of OTOF gene of one family patient were PCR amplified using specific primers. Each fragment was purified and subsequently analyzed by direct sequencing in an Applied Biosystems 3 730 automated DNA sequencer. The resultant sequence data were compared with the standard sequence to identify deafness-associated mutations. Other DNA samples were then screened for these mutations by PCR amplification and sequence analysis.PCR amplifications were successfully conducted in all the subjects. Comparison of the resultant OTOF sequence in one family patient with the standard sequence identified 10 nucleotide variants which do not lead to amino acid change. These mutations were also detectable in other family individuals, 3 sporadic AN patients and 3 normal-hearing controls.The OTOF does not seem to contribute to the pathogenesis of this Chinese AN family, which suggest new gene(s) involvement.

The tomato ripening mutant, ripening inhibitor (rin), whose fruits fails to ripen, has been identified and widely studied. The RIN gene has been cloned. Here we present the expression of a truncated form of the RIN protein from tomato and the preparation of a polyclonal antibody against it. The resulting antibody recognized the RIN of crude protein extracts from different tomato tissues. The protein level of RIN in tomato was detected with this antibody by western blot, which suggested the accumulation of RIN protein increased gradually during tomato fruit ripening.

We explored the clinical and molecular characterization of a Chinese family with non-syndromic hearing impairment. Clinical evaluations revealed a possible maternal inheritance pattern, and showed an extremely similar phenotype of hearing loss including the age of onset, severity, and audiometric configuration. Sequence analysis of the mitochondrial 12S rRNA and tRNA(Ser(UCN)) genes led to the identification of a homoplasmic A827G mutation in all maternal relatives, which was absent in other family members and 40 Chinese controls. This mutation has previously been reported sporadically in a few individuals with aminoglycoside-induced and non-syndromic hearing loss. The A827G mutation is located at the A-site of the mitochondrial 12S rRNA gene which is highly evolutionarily conserved in mammals. The occurrence of the A827G mutation in these genetically unrelated subjects strongly suggests that this mutation is involved in the pathogenesis of hearing impairment. However, incomplete penetrance of hearing loss indicates that the A827G mutation alone is not sufficient to produce clinical phenotype but requires the involvement of modifier factors for the phenotypic expression, even though aminoglycosides and GJB2 gene may not contribute to the penetrance of the A827G mutation in this Chinese family. In contrast with the variable phenotype of hearing loss associated with other mitochondrial mutations, all of the patients in our family exhibited strikingly similar clinical features. This discrepancy likely reflects the difference of genetic backgrounds between this pedigree and others.

The process for Czochralski growth of convex interface Nd:YAG laser crystal with [111] seed was studied. The forming mechanisms of central core and lateral core through growth of facets were discussed. The relations between the shape of solid/liquid interface and temperature gradient in the melt and facets' growth were described. The technical methods of elimination of lateral core and reduction of central core were achieved. The good results of growth of lateral core-free and smaller central core Nd:YAG crystals were got through designing apparatus and temperature field to maintain steady growth interface pattern.© (1996) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.