Search

Background: Chemokine 13 (CXCL13) and its chemokine receptor 5 (CXCR5) are involved in the onset of various types of cancer. However, their role in cervical cancer (CC) remains unknown. Objective: To investigate the role of chemokine 13 (CXCL13) and its receptor in CC. Methods: The expression of CXCL13/CXCR5 and the infiltration of CXCR5+CD8+ T cells in CC, cervical intraepithelial neoplasia (CIN), normal cervical epithelial (NCE) tissues, and in CC cell lines were analysed and the associated clinical significance was determined. In vitro, CXCL13 overexpression and DNA methyltransferase inhibition (through S110) were used to investigate the biological function and the underlying mechanism that regulates CXCL13 expression. Tumor growth and liver metastasis were also evaluated in the xenogenous subcutaneously implant model. Results: CXCL13/CXCR5 expression levels and the infiltration of CXCR5+CD8+ T cells were significantly decreased in CC tissues compared with CIN and NCE tissues. CXCL13 downregulation was significantly correlated with the FIGO stages, lymph node metastasis, interstitial infiltration depth, and pathological grade. The overexpression of CXCL13 suppressed CC cell migration. CXCL13 downregulation was associated with hypermethylation in CC cell lines, and primary tumor biopsies. Furthermore, a CpG dinucleotide at the HIF-1a transcription factor motifs in the promoter element of CXCL13 was consistently methylated in CC cells and associated with HIF-1a. CXCL13 overexpression and S110 treatment dramatically repressed tumor growth and liver metastasis in the xenograft model; whereas it’s low expression increased the risk of death in CC patients. Conclusion: DNA methylation-dependent CXCL13 downregulation may promote cervical carcinogenesis and progression.

Egg fertilization is a dynamic process, including varieties of biochemical changes. To better understand the molecular mechanisms during the egg embryo development, the objective of this study was to quantify protein expression changes between fertilized and unfertilized Beijing-You chicken eggs using label-free liquid chromatography-tandem mass spectrometry method. The results showed that a total of 1241 proteins were identified from fertilized and unfertilized eggs, 229 proteins were observed difference in fertilized eggs (p

Our previous study demonstrated that intranasal administration of histone deacetylase inhibitor sodium butyrate (NaB) exhibits therapeutic effects on a mouse model of allergic rhinitis (AR). However, whether NaB is effective on AR when administered orally and prophylactically, as well as its potential effects on gene expression, remained unknown. The present study aimed to investigate the preventive effect of NaB on AR when added to the diet of newly weaned mice and to evaluate the changes in long non‑coding (lnc)RNA and mRNA expression profiles in the nasal mucosa. Mice were randomly divided into three groups as follows: i) Control (C) group, (no treatment); ii) AR group [treated with ovalbumin (OVA)]; and iii) NaB + AR group (treated with OVA and NaB). The NaB + AR group was administered NaB in their feed (30 g/kg chow), whereas the other two groups were fed normal feed between 3 and 6 weeks of age. At 7 weeks of age, OVA administration was initiated to induce AR in the AR and NaB + AR groups. Following model establishment, behavioral assessments, western blotting and gene expression analysis were performed. NaB exhibited a preventive effect in the murine AR model, diminished the increases in histone deacetylase 1 (HDAC1) and HDAC8 expression and increased OVA‑induced acetylation of histone H3 at lysine 9. In addition, NaB increased the AR‑associated low expression of interleukin 2 (IL‑2), interferon γ and IL‑17 and decreased the expression of IL‑4, IL‑5 and transforming growth factor β1. Gene Ontology and pathway analyses revealed the top 10 pathways among the groups. Octamer‑binding transcription factor 1, ecotropic viral integration site 1 and paired box 4 were predicted to be target genes of lncRNA (NONMMUT057309). Thus, NaB may exhibit a preventive effect on AR. Additionally, the lncRNA and mRNA expression profiles in the nasal mucosa of mice with AR differed significantly following NaB treatment. These results may provide insights into the pathogenesis of AR and suggest new treatment targets.

BACKGROUND Dendrobium is a genus of plants used as traditional Chinese herbal medicines, with high economic and medicinal value. METHODS AND RESULTS To reveal the mechanism of flavonoid biosynthesis in Dendrobium, the metabolites and transcriptomes of four Dendrobium species (D. chrysotoxum, D. nobile, D. fimbriatum, and D. denneanum) were analyzed comprehensively. Ultra-high-performance liquid chromatography-tandem mass spectrometry analysis revealed ten flavonoid compounds in Dendrobium. In total, 100,096 unigenes were obtained from the transcript database of the four Dendrobium species. Among the identified differentially expressed genes, 51 were associated with flavonoid biosynthesis, and 670 differentially expressed transcription factors were predicted, including 194 MYB, 87 bHLH, and 100 WRKY family transcription factors, respectively. Transcriptome analysis showed that the expression levels of structural genes such as chalcone synthase (CHS), cinnamate-4-hydroxylase (C4H), and flavonoid 3'-hydroxylase (F3'H) were lower in D. chrysotoxum, D. nobile, and D. fimbriatum than those in D. denneanum, which may be the main reason for the low flavonoid contents in D. chrysotoxum, D. nobile, and D. fimbriatum. CONCLUSIONS The expression level of structural genes corresponded to the accumulation level of flavonols in the different Dendrobium species. The results deepen the understanding of the molecular mechanism of flavonoid biosynthesis in Dendrobium and provide novel insights into the synthesis and accumulation of flavonoids in Dendrobium.

Dendrobium is known for its pharmacological actions including anti-cancer effect, anti-fatigue effect, gastric ulcer protective effect, and so on. At present, only studies on endophytic fungi of Dendrobium affecting the metabolites of host plants have been reported, very little research has been done on endophytic bacteria. In this study, we have demonstrated the great diversity of endophytic bacteria in 6 Dendrobium samples from different origins and cultivars. According to the results of the culture-independent method, the endophytic bacterial community in Dendrobium stems showed obvious different in the 6 samples and was influenced by origin and cultivar. Some bacteria including Ralstonia, Comamonas and Lelliottia were first detected in Dendrobium in this study. Based on the culture-dependent method, a total of 165 cultivable endophytic bacteria isolates were isolated from the sterilized Dendrobium stems, and were classified into 43 species according to the 16S rRNA gene sequence analysis. Moreover, 14 of the 43 strains showed antimicrobial activity against phytopathogen using the Kirby-Bauer method. Strain NA-HTong-7 (Bacillus megaterium, 99.12%) showed the highest antimicrobial activity. This study was the first comprehensive study on endophytic bacteria of Dendrobium from different origins and cultivars, which provides new insights into the endophytic bacteria from Dendrobium.

Fresh kidney beans (Phaseolus vulgaris L.) are common green vegetable that are widely cultured and consumed worldwide. The benefits of fresh kidney beans are highly substantial, but at the same time, toxicity caused by phytohemagglutinin (PHA) has raised concerns. PHA from fresh kidney beans accounted for 40% of poisoning events induced by poisonous plants in China from 2004 to 2013. However, an understanding of PHA levels in fresh kidney beans remains elusive. An analysis method for PHA based on agglutination of PHA to erythrocyte cell was established. We investigated the effect of cultivar, bean part, maturity, and cooking method on PHA content in fresh kidney beans. We demonstrated significant differences in the distribution of PHA among various predominant, fresh Chinese kidney bean cultivars and parts. One major Chinese strain, Zihuayoudou, was found to contain the highest amount of PHA, which was abundant in fresh kidney bean seeds. Moreover, PHA concentrations were found to decrease with the maturity of the fresh product. Further, cooking methods had a significant effect on PHA levels in fresh kidney beans. PHA could be effectively removed when stir-frying for more than 18 min, or braising for more than 10 min. These findings provide insight into the PHA content of fresh kidney beans in China and will be helpful to guide food safety measures.

Dendrobium candidum is used as a traditional Chinese medicine and as a raw material in functional foods. D. candidum stems are green or red, and red stems are richer in anthocyanins. Light is an important environmental factor that induces anthocyanin accumulation in D. candidum. However, the underlying molecular mechanisms have not been fully unraveled. In this study, we exposed D. candidum seedlings to two different light intensities and found that strong light increased the anthocyanin content and the expression of genes involved in anthocyanin biosynthesis. Through transcriptome profiling and expression analysis, we identified a WD40-repeat transcription factor, DcTTG1, whose expression is induced by light. Yeast one-hybrid assays showed that DcTTG1 binds to the promoters of DcCHS2, DcCHI, DcF3H, and DcF3′H, and a transient GUS activity assay indicated that DcTTG1 can induce their expression. In addition, DcTTG1 complemented the anthocyanin deficiency phenotype of the Arabidopsis thaliana ttg1-13 mutant. Collectively, our results suggest that light promotes anthocyanin accumulation in D. candidum seedlings via the upregulation of DcTTG1, which induces anthocyanin synthesis-related gene expression.

Dendrobium candidum stems are used as Chinese medicine and functional food. Red stems of D. candidum are rich in anthocyanins, which attract pollinator insects, protect the plants against environmental stress, and improve human health. The regulatory mechanisms of anthocyanin biosynthesis and stem color differentiation in D. candidum are not fully understood. Using transcriptome profiling, we identified a basic helix-loop-helix transcription factor (DcTT8) involved in anthocyanin biosynthesis in D. candidum stems. Ultraperformance liquid chromatography-tandem mass spectrometry was used to determine pigment contents and compositions in red and green stems, revealing that cyanidin is responsible for the red color. DcTT8 could bind the DcF3'H and DcUFGT promoters and finely regulate DcF3'H and DcUFGT expression. Our data indicate that DcTT8 participates in anthocyanin biosynthesis and offers novel insights into the transcriptional regulation of anthocyanin biosynthesis in D. candidum.

As an important global crop, the potato (Solanum tuberosum L.) contains the endotoxin solanine that leads to human poisoning and major economic losses. Poisoning symptoms and even acute poisoning may occur when the content of solanine in potatoes exceeds 200 mg/kg. In addition, potatoes are susceptible to some pathogenic bacteria, including Streptomyces scabies and Erwinia carotovora subsp. atroseptica (Van Hall) dye, which can cause potato scab and potato blackleg disease, respectively. In this study, 37 culturable endophytic bacteria strains were obtained from potato tubers based on the culture-dependent method. Results indicated that nine strains showed antimicrobial activity against at least one pathogen by antimicrobial activity screening and 23 strains showed inhibitory activity on solanine in potato tubers. Among them, strain P-NA2-14 (Bacillus megaterium NBRC 15308T, 99.31%) showed not only better antimicrobial activity against both the two indicator pathogens, but also the best inhibitory activity on solanine, which was proved to be a potential biocontrol bacterium. Meanwhile, the relationship between the distribution of the endophytic bacterial community and the content of solanine in potato tubers was studied by Illumina-based analysis, indicating that the distribution of the endophytic bacterial community was obviously influenced by the content of solanine. The results showed a new insight into the relationship between plant secondary metabolites and endophytic bacteria in potato tubers and provided potential new technical support for the biological control of potato storage.

Human gut bacteria contribute significantly to human health and several studies have evaluated the effects of dietary fibers on human gut bacterial ecology. However, the relationship between different degrees of fiber polymerization and human gut bacteria is unknown. Here, we analyzed three fiber substrates with different degrees of polymerization, namely carboxymethylcellulose, β-glucans, and galactooligosaccharides. To probe the in vitro influence of the degree of polymerization of the fiber on human gut bacteria, we measured the pH, air pressure, and short-chain fatty acid content of fecal fermentation supplemented with these fiber substrates, and sequenced the 16S ribosomal RNA genes of the microbial community in the fiber-treated fermentations. The butyric acid concentration was shown to decline with decreasing degree of polymerization of the fiber. Illumina Miseq sequencing indicated that the degree of polymerization might have an influence on human gut microbial diversity and abundance. Principal coordinate analysis unveiled a relationship between the degree of fiber polymerization and the gut bacterial community. Specific microbiota operational taxonomic units (OTUs) within the genera Escherichia-Shigella, Fusobacterium, and Dorea were proportional to the degree of fiber significantly, whereas OTUs within the genera Bifidobacterium, Streptococcus, and Lactobacillus were inversely correlated with the degree of polymerization. Correlation analysis between the fiber degree of polymerization and gut bacteria may demonstrate the effect of fibers on gut microbiota, and subsequently, on human health.

Dietary consumption of selenium-rich agro-food is an effective way to avoid selenium deficiency diseases, however, over consumption of selenium-rich agro-food will result in potential risk of selenosis and problems with associated metals. In this study, we measured the concentrations of selenium and its associated metals in 2756 common and 4894 selenium-rich agro-food samples in 10 regions of China. We found that selenium-rich rice, flour, edible fungi and algae, meat, and tea contain higher levels of associated metals than other selenium-rich agro-food samples. Increasing the consumption of selenium-rich agro-food could make the actual intakes (AIs) of selenium for all population to meet respective recommended daily intakes (RDIs). Benefit-risk assessment results indicated that increasing the consumption of selenium-rich agro-food make AIs of selenium for all populations meet RDIs, chromium intakes for people under 18 years old exceed provisional tolerated daily intake (PTDIs), while arsenic and cadmium intakes are close to PTDIs. The main dietary contributors of selenium, chromium, arsenic and cadmium were meat, edible fungi and algae, rice, and rice, respectively. The study supported the consumption of selenium-rich agro-food for effective selenium supplement, but also emphasized potential risk from associated metals in selenium-rich agro-food, especially chromium.

A robust and high-throughput method was developed for the determination of 108 pesticide residues in Traditional Chinese Medicines (TCMs) simultaneously using a combination of UHPLC-MS/MS analysis and the modified QuEChERS method. Extraction was carried out in acetonitrile containing 0.75% (v/v) acetic acid with ultrasonication for 15 min; MgSO4 and C18 were used as the dispersive-solid phase extraction sorbents. The method exhibited good linearity (r2 > 0.9901), in addition to good selectivity, precision and repeatability. More than 92% of pesticides exhibited high rates or recovery in the 70–120% range. This method showed high sensitivity, with Limits of Quantitation in the 0.01–20 ng/mL range in Cortex Moutan, and 0.01–50 ng/mL in the other TCMs. The method was employed for the analysis of 39 real samples from different habitats, and pesticides were detected in 92.3% of the samples, with 26 pesticides being detected in these three TCMs. More than four pesticides were detected in a third of the samples. Among them, tebuconazole was detected in all the three TCMs with 0.22–22.02 μg/kg concentration, which was lower than the provisions in GB 2763-2019 (50 μg/kg). In addition, the paclobutrazol detection rate in Ophiopogon japonicus was 100%, and the detected concentrations of 9 samples exceeded the Maximum Residue Levels defined for vegetables (50 μg/kg). Considering there are no regulations that govern the limits of pesticide residues in the three TCMs in China, we recommend the acceleration of efforts to introduce appropriate regulations.

Lignin is one of the most important components of the plant cell wall, and the expression and transcriptional regulation of lignin biosynthesis-related genes have been studied widely in Arabidopsis and other plants. Citrus fruit juice sacs often undergo lignification, particularly during fruit ripening and storage periods; however, the underlying genetic mechanisms have been little investigated. In this study, we isolated and identified CsMYB330 and CsMYB308 transcription factors, and found that their expression levels are significantly altered during the lignification of citrus fruit juice sacs. We found that CsMYB330 and CsMYB308 can recognize and bind AC elements in the Cs4CL1 promoter and finely regulate expression of the Cs4CL1 gene. In this regulatory process, CsMYB330 was identified as a transcriptional activator, whereas CsMYB308 appears to be a transcriptional repressor. In addition, using a transient assay, we demonstrated that expression of the Cs4CL1 gene is significantly altered in fruit juice sacs overexpressing these two transcription factors. These results indicate that the transcription factors CsMYB330 and CsMYB308 play important roles in the lignification of citrus fruit juice sacs and provide novel insights into the transcriptional regulation associated with fruit juice sac lignification.

Sleep deprivation (SD) negatively caused cognitive deficit, which was associated with oxidative stress induced damage. Ginsenoside Rh2 had the ability to protect against damage caused by reactive oxygen species in vitro, showing antioxidant property. Therefore, it was hypothesized that Ginsenoside Rh2 could prevent SD-induced cognitive deficit via its antioxidant properties. In this study, the effect of Ginsenoside Rh2 on memory impairment induced by sleep deprivation was investigated. The mice were sleep deprived continuously for 14 days using our self-made Sleep Interruption Apparatus (SIA). Ginsenoside Rh2 was administered intraperitoneally at two doses (20 and 40 μmol/kg) for 20 days. Thereafter, behavioral studies were conducted to test the learning and memory ability using object location recognition (OLR) experiment and passive avoidance (PA) test. Additionally, the oxidative stress parameters in the serum and the brain tissues (cortex and hippocampus) were assessed, including the superoxide dismutase (SOD) enzyme activity, the total antioxidant reactivity (TAR), the malondialdehyde (MDA) level, the glutathione (GSH) level, and the lipid peroxidation (LPO) content. The results revealed that SD impaired both spatial and non-spatial memory (P < 0.05). Treatment with Ginsenoside Rh2 at both doses prevented memory impairment induced by SD. Moreover, Ginsenoside Rh2 normalized the reduction of SOD and TAR activities in the serum (P < 0.01) and the decrease of GSH content in both the cortex and hippocampus (P < 0.05) induced by SD. Furthermore, Ginsenoside Rh2 significantly decreased the MDA level in the serum (P < 0.05) and the LPO content in both the cortex and hippocampus (P < 0.05) compared to SD group. In conclusion, sleep deprivation impaired both spatial and non-spatial memory and Ginsenoside Rh2 reversed this impairment, probably by preventing the oxidative stress damage in the body, including the serum and brain during sleep deprivation.

Sleep deprivation (SD) is associated with oxidative stress that causes learning and memory impairment. 20(S)-Protopanaxadiol (PPD), one of the protopanaxadiol-type saponins, has antioxidant and neuroprotective effect. This study was designed to research the protective effect of PPD against cognitive deficits induced by chronic sleep deprivation (CSD) in mice. The CSD model was induced by subjecting the mice to our self-made Sleep Interruption Apparatus (SIA) continuously for 14 days. The memory enhancing effects of PPD were evaluated by behavioral tests and the related mechanism was further explored by observing the oxidative stress changes in the cortex and hippocampus of mice. The results revealed that PPD (20 and 40 μmol/kg, i.p.) administration significantly improved the cognitive performance of CSD model mice in object location recognition experiment, novel object recognition task and Morris water maze test. Furthermore, PPD effectively restored the levels/activities of antioxidant defense biomarkers in the cortex and hippocampus, including the superoxide dismutase (SOD) enzyme activity, catalase (CAT) enzyme activity, glutathione (GSH), and lipid peroxidation (LPO). In conclusion, PPD could attenuate cognitive deficits induced by CSD, and the neuroprotective effect of PPD might be mediated by alleviation of oxidative stress. It was assumed that PPD has the potential to be a neuroprotective substance for cognition dysfunction.

The amino acid permease (AAP) is an important transmembrane protein that is involved in the absorption and transport of amino acids in plants. We investigated the expression patterns of AtAAP genes in Arabidopsis thaliana, based on quantitative real-time PCR. The results revealed differential expression patterns of eight AtAAP genes in different tissues, with five genes (AtAAP1, AtAAP2, AtAAP6, AtAAP7, and AtAAP8) expressed at relatively high levels in both flowers and siliques, suggesting their shared functions in the accumulation of amino acids. In transgenic plants, with endosperm-specific overexpression of AtAAP1, both AtAAP1 and AtAAP6 were up-regulated in both the roots and siliques, while AtAAP2, AtAAP3, AtAAP4, and AtAAP5 showed similar expression levels in the stems and siliques, whereas AtAAP7 and AtAAP8 were expressed at their highest levels in the stems and roots. The results of the amino acid affinity experiments revealed varied absorption capacities for different amino acids, by AtAAP1, and increased acid amino contents in the reproductive organs. These results were verified in transgenic maize plants, with the overexpression of AtAAP1, revealing higher amino acid contents in the reproductive organs than those of the vegetative organs. Our study clearly demonstrated that the endosperm-specific promoter increased the amino acid contents in the reproductive organs and improved the effective utilization of organic nitrogen in plants.

Congenital cataract (CC) is a clinical and genetically heterogeneous eye disease that primarily causes lens disorder and even amblyopic blindness in children. As the mechanism underlying CC is genetically inherited, identification of CC-associated gene mutations and their role in protein distribution are topics of both pharmacological and biological research. Through physical and ophthalmic examinations, two Chinese pedigrees with autosomal dominant congenital cataract (ADCC) were recruited for this study. Mutation analyses of CC candidate genes by next-generation sequencing (NGS) and Sanger sequencing revealed a novel missense mutation in CRYBB2 (p.V146L) and a deletion mutation in CRYAA (p.116_118del). Both mutations fully co-segregated were not observed in unaffected family members or in 100 unrelated healthy controls. The CRYBB2 missense mutation disrupts the distribution of CRYBB2 in human lens epithelial cells (HLEpiCs), and the CRYAA deletion mutation causes hyperdispersion of CRYAA. Furthermore, these two crystallin mutations result in aberrant expression of unfolded protein response (UPR) marker genes as well as apoptosis in HLEpiCs. Collectively, these findings broaden the genetic spectrum of ADCC.

The objective of this study was to determine the effects of low-concentration acidic electrolyzed water (LcEW) combined with ultrasound (US) on fresh-sliced button mushrooms (Agaricus bisporus), by evaluating the enzymatic browning, physio-biochemical changes, and microbial loads. The sliced button mushrooms were divided randomly into three groups (control, LcEW, and LcEW+US) and treated separately with tap water, LcEW, and LcEW+US for 3 min, and afterwards, they were stored at 5 °C for 8 days. The results indicated that LcEW delayed surface browning and maintained the flesh firmness of fresh-cut mushrooms compared with the control, and ultrasound enhanced these effects. Moreover, LcEW+US slowed down the electrolyte leakage (EL) rate and malondialdehyde (MDA) content, which were 5.29% and 0.5227 μmol/fresh weight less than those in control at day 8, respectively. The combined treatment of LcEW and US also significantly inhibited the activities of polyphenol oxidase (PPO) and peroxidase (POD) and controlled the counts of total bacteria (TBC) and yeast and mold counts (Y&M). Meanwhile, LcEW+US maintained a high level of total soluble protein and total phenolic contents, which were 1.49 and 1.24 times higher than those in control at the end of the storage time, respectively, and the combined treatment was more effective than LcEW treatment alone. These results demonstrated that the combination of LcEW and US could be an effective pretreatment technology in maintaining the product quality and prolonging the shelf life of fresh-sliced button mushrooms.

Autosomal dominant congenital cataract (ADCC), the most common hereditary disease, is a major cause of eye disease in children. Due to its high genetic and clinical heterogeneity, the identification of ADCC-associated gene mutations is essential for the development of molecular therapies. In this study, we examined a four-generation Chinese pedigree with ADCC and identified putative mutations in ADCC candidate genes via next-generation sequencing (NGS) followed by Sanger sequencing. A novel missense mutation in GJA8 (c.T217C) in ADCC patients causes a serine-to-proline substitution at residue 73 of connexin 50 (Cx50); no mutation was found in unaffected family members and unrelated healthy individuals. Functional analysis revealed that this missense mutation disrupts protein function in human lens epithelial cells (HLEpiCs), which fails to form calcium-sensitive hemichannels. Furthermore, mutant Cx50 leads to decreased ROS scavenging by inhibiting G6PD expression and thus induces cell apoptosis via aberrant activation of the unfolded protein response (UPR). In conclusion, we report a novel GJA8 heterozygous mutation in a Chinese family with a vital role in ADCC, broadening the genetic spectrum of this disease.

Background Embryo chicken egg is a nutritional supplement that has been used to enhance physical fitness and promote wound healing according to traditional Chinese medicine for many years. In this study, we evaluated the effects of embryo chicken egg extract (ECE) on the exercise performance and fatigue in mice and the underlying mechanisms. Results The results indicated that ECE can prolong the exhaustive swimming time, decrease lactic acid, blood urea nitrogen, creatine kinase, and malondialdehyde levels, and increase superoxide dismutase, glutathione peroxidase, and glycogen levels. Additionally, ECE can also regulate the balance of oxidative stress via the adenosine monophosphate activated protein kinase/mammalian target of rapamycin signalling pathway. Conclusion Taken together, these results showed that ECE can improve exercise performance and reduce physical fatigue in mice, which indicates that ECE can be used as a potential supplement to reduce physical fatigue. © 2020 Society of Chemical Industry.

Sleep deprivation has been widely reported to cause cognitive dysfunction, and elevation in oxidative stress and inflammation in the body, including the brain, have been suggested as the main factors. Genistein (GE) is an isoflavone widely present in leguminous plants, and it was found to exert a wide spectrum of biological activities, including antioxidant, anti-inflammatory, hepatoprotective, neuroprotective, and antimetastatic effects. In this study, the protective effect of GE on chronic sleep deprivation (CSD)-induced cognitive dysfunction was investigated. The mice were subjected to the sleep interruption apparatus and continuously sleep deprived for 25 days. GE was orally administrated (10, 20, and 40 mg/kg) during the sleep deprivation process totally for 25 days. Cognitive behavioral tests were conducted to study the learning and memory using the object location recognition (OLR) task, novel object recognition (NOR) test, and the Morris water maze (MWM) task. Additionally, the cortex and hippocampus were dissected to measure the oxidative stress markers and the antioxidant element nuclear erythroid-2-related factor 2 (Nrf2) and its downstream targets, including glutamate cysteine ligase catalytic, glutamate cysteine ligase modifier, heme oxygenase 1, and quinone oxidoreductase 1, as well as nuclear factor kappa B (NF-κB) p65, nitric oxide synthase (iNOS), and cyclooxygenase 2 (COX-2) protein expression. Moreover, the pro-inflammatory cytokines (TNF-α, interleukin [IL]-6, and IL-1β) level was examined in the serum. The current results showed that GE could dose-dependently ameliorate the cognitive deficits of CSD-treated mice in the OLR, NOR, and MWM tasks. In addition, GE treatment significantly elevated the activities of total antioxidant capacity and superoxide dismutase and the level of glutathione and lowered the content of malondialdehyde in the cortex and hippocampus of CSD-treated mice. Furthermore, GE administration effectively activated the antioxidant element Nrf2 and its downstream targets in the cortex and hippocampus of CSD-treated mice. Moreover, GE treatment significantly suppressed CSD-induced NF-κB p65, iNOS, and COX-2 activation in the cortex and hippocampus, as well as inhibited CSD-induced pro-inflammatory cytokines (TNF-α, IL-6, and IL-1β) release in the serum. Taken together, all these results suggested that GE has substantial potential as a therapeutic intervention for the alleviation of CSD-induced deleterious effects.

Varieties of Citrus are commercially important fruits that are cultivated worldwide and are valued for being highly nutritious and having an appealing flavor. Lignification of citrus fruit juice sacs is a serious physiological disorder that occurs during postharvest storage, for which the underlying transcriptional regulatory mechanisms remain unclear. In this study, we identified and isolated a candidate MYB transcription factor, CsMYB85, that is involved in the regulation of lignin biosynthesis in Citrus sinensis, which has homologs in Arabidopsis and other plants. We found that during juice sac lignification, CsMYB85 expression levels increase significantly, and therefore, suspected that this gene may control lignin biosynthesis during the lignification process. Our results indicated that CsMYB85 binds the CsMYB330 promoter, regulates its expression, and interacts with CsMYB308 in transgenic yeast and tobacco. A transient expression assay indicated that Cs4CL1 expression levels and lignin content significantly increased in fruit juice sacs overexpressing CsMYB85. At4CL1 expression levels and lignin content were also significantly increased in Arabidopsis overexpressing CsMYB85. We accordingly present convincing evidence for the participation of the CsMYB85 transcription factor in fruit juice sac lignification, and thereby provide new insights into the transcriptional regulation of this process in citrus fruits.

Juice sac granulation leads to large postharvest losses of citrus fruits. Although the fundamental mechanism of this process remains not fully understood, some previous studies have clearly shown the close association between lignin metabolism and juice sac granulation. While miRNAs have been proved as main regulators of target gene expression which regulates lignin biosynthesis in some plant species, the role of miRNAs in citrus granulation was still unknown. To evaluate the role of miRNAs in granulation, homology search-based computational approach was employed to identify miRNAs in citrus, and 12 potentially unreported miRNAs in citrus were obtained. Through the identification and annotation of target genes, two miRNA families (miR397, miR828) were shown to be involved in lignin biosynthesis pathways. The expression pattern of miRNAs at different granulation levels was further investigated by real-time quantitative reverse transcription PCR (qRT-PCR). The results demonstrated that the expression of csi-miR397 and N-miR828 were negatively related to citrus granulation, with correlation coefficients of 0.888 and 0.944, respectively. A further study on their targets by qRT-PCR revealed the expressions of Cs6g06890.1 and Cs1g17590.1 were positively correlative to granulation, with correlation coefficients of 0.917 and 0.963, respectively. The correlation analysis among the expression of miRNA, its target, lignin content and granulation degree indicates a likely role of the csi-miR397/Cs6g06890.1 and N-miR828/Cs1g17590.1 modules were correlate to granulation through regulation of the internal lignin activity. The results of this study lay a foundation for miRNAs in regulating lignin processes which relate to citrus granulation on post-transcriptional level.

Numerous studies have focused on the effects of food-processing techniques on pesticide residues. However, it remains a challenge to ensure high-quality processing and effective pesticide removal from foods. Triadimefon (TF) is a broad-spectrum fungicide that is widely used in barley crop, and its residues are detected in its products, including beer. In this study, we investigated the dissipation kinetics of TF during fermentation mediated by two different yeast strains, Saccharomyces cerevisiae IAPPST 1401 (Y1) and CICC 1202 (Y2), and found that Y2 promoted the degradation of TF. Response surface methodology was used to optimize fermentation process variables, in order to achieve the maximum removal rate of TF and the minimum production of its corresponding metabolite, triadimenol (TN). Processing factors (PFs) were also evaluated during the optimized brewing process and were close to 1 for TF during the malting, milling, boiling, and cooling steps, but not in mashing and fermentation that were 0.19 and 0.13, respectively. TF degraded to TN during brewing, and a PF value of >1 for TN was also observed in malting and fermentation. Our analysis concluded that beers produced using the yeasts and brewing methods we investigated are safe for human consumption.

The synthesis of a novel sugar coated poly( p -phenyleneethynylene) (PPE) via Pd-catalyzed Sonogashira reaction and its subsequent interaction with concanavalin A (Con A) are described. UV–Vis and fluorescence spectra were employed to investigate the response of the polymer toward various concentrations of Con A. The polymer showed a green fluorescence which can be quenched by Con A due to complex formation at ground state. The protocols reported herein provide intermediate products that can be converted into useful sensing elements from cheap and commercially available starting materials, in a few number of steps.

Microbial fermentation can endow food with unique flavors, increase its nutritional value and enhance functional characteristics. Our previous research has shown that liquid fermentation of soymilk by Bacillus subtilis BSNK-5 imparted new functional properties of to the fermented product via production of nattokinase. In this study, in order to further investigate the changes in the flavor, nutritional quality and functional characteristics of soymilk during fermentation using proton nuclear magnetic resonance (1H NMR) metabolomics to monitor the metabolite profile of BSNK-5-fermented soymilk. A total of 44 differential metabolites were identified between BSNK-5-fermented soymilk and uninoculated/unfermented soymilk, among which the levels of flavor-related substances (acetate, isovalerate and 2-methylbutyrate), nutrient-related substances (12 free amino acids), and functional substances (taurine, GABA and genistein) significantly increased after fermentation. These metabolites were closely associated with eight potential metabolic pathways. This work highlighted the significance of BSNK-5 strain in improving the nutritional quality and functional characteristics of fermented soymilk; however, the use of the strain also caused flavor deterioration. This study lays a theoretical foundation for the improvement and development of fermented soy products via liquid fermentation with B. subtilis.

Genistein (GE) was reported to exert a wide spectrum of biological activities, including antioxidant, anti-inflammatory, anti-mutagenic, anticancer, and cardio-protective effects. In addition, both clinical and preclinical studies have recently suggested GE a potential neuroprotective and memory-enhancing drug against neurodegenerative diseases. The animal model of scopolamine (Scop)-induced amnesia is widely used to study underlying mechanisms and treatment of cognitive impairment in neurodegenerative diseases. However, there is no report about the effects of GE on Scop-induced amnesia in mice. Therefore, the present study was carried out to investigate the beneficial effects and potential mechanism of GE against Scop-induced deficits in mice. The mice were orally pretreated with either GE (10, 20, and 40 mg/kg) or donepezil (1.60 mg/kg) for 14 days. After the pretreatment, the open field test was conducted to assess the effect of GE on the locomotor activity of mice. Thereafter, mice were daily injected with Scop (0.75 mg/kg) intraperitoneally to induce memory deficits and subjected to the cognitive behavioral tests including the Object Location Recognition (OLR) experiment and Morris Water Maze (MWM) task. After the behavioral tests, biochemical parameter assay and western blot analysis were used to examine the underlying mechanisms of its action. The results showed that GE administration significantly improved the cognitive performance of Scop-treated mice in OLR and Morris water maze tests, exerting the memory-enhancing effects. Additionally, GE remarkably promoted the cholinergic neurotransmission and protected against the oxidative stress damage in the hippocampus of Scop-treated mice, as indicated by decreasing AChE activity, elevating ChAT activity and Ach level, increasing SOD activity, lowering the level of MDA and increasing GSH content. Furthermore, GE was found to significantly upregulate the expression levels of p-ERK, p-CREB and BDNF proteins in the hippocampus of Scop-treated mice. Taken together, these results for the first time found that GE exerts cognitive-improving effects in Scop-induced amnesia and suggested it may be a potential candidate compound for the treatment of some neurodegenerative diseases such as Alzheimer's Disease (AD).

In the recent years, interest in soybean as a neuroprotective nutrient in the management of Alzheimer&rsquo, s disease (AD) has increased and soy isoflavones (SI), as kinds of soybean phytochemicals, are thought to be biologically active components that confer this beneficial effect against neurodegenerative diseases. However, the neuroprotective effect of SI is not well understood. Therefore, the present study (30 days) was conducted to investigate the neuroprotective effects of soy isoflavones (SI) on scopolamine (SCOP)-induced memory impairments in Institute of Cancer Research (ICR) mice (aged 4 weeks) and to elucidate its underlying mechanisms of action. SI (40 mg/kg) administration improved the cognitive performance of SCOP-treated mice in an object location recognition task and the Morris water maze test. SI (40 mg/kg) administration significantly enhanced cholinergic system function and suppressed oxidative stress levels in the hippocampus of SCOP-treated mice. Furthermore, SI (40 mg/kg) treatment markedly upregulated the phosphorylation levels of extracellular signal-regulated kinase (ERK), cAMP response element-binding protein (CREB) and brain-derived neurotrophic factor (BDNF) expression levels in the hippocampus. Taken together, these results demonstrated that soy isoflavones exerted a significant neuroprotective effect on cognitive dysfunctions induced by scopolamine, suggesting that soy isoflavones could be a good candidate for possible treatment of neurodegenerative diseases, such as Alzheimer&rsquo, s disease (AD).

Response surface methodology (RSM) was employed to optimize the conditions for the ultrasonic-assisted extraction (UAE) of polysaccharides from the flowers of Dendrobium devonianum. The optimal conditions for the maximum yields of DDFPs are as follows: an extraction temperature of 63.13°C, an extraction time of 53.10 min, and a water-to-raw material ratio of 22.11 mL/g. Furthermore, three fractions (DDFPs30, DDFPs50, and DDFPs70) were prepared from Dendrobium devonianum flowers polysaccharides (DDFPs) by the stepwise ethanol precipitation method. The DDFPs50 exhibited the highest antioxidant activity compared to the other fractions. The molecular weight, polydispersity, and conformation of these fractions were also characterized. In particular, the monosaccharide composition analysis of the DDFPs indicates that mannose and glucose are the primary components, similar to those of the D. officinale plant. This study provides a rapid extraction technology and essential information for the production of DDFPs, which could be potentially used as healthcare food.

Background: Ginsenoside Rh1, a 20(S)-protopanaxatriol with a single sugar moiety from ginseng, has benefits on the central nervous system and recently Rh1 had been reported to be effective for the Alzheimer’s disease (AD). Objective: The present study was conducted to investigate the neuroprotective effects of Ginsenoside Rh1 on scopolamine-induced cognitive dysfunctions and to elucidate its underlying mechanisms of action. Methods: Rh1 (20 and 40 μmol/kg) was administered intraperitoneally to mice for 28 days, and scopolamine (0.75 mg/kg) was injected intraperitoneally for 10 days to induce memory deficit. Memory-related behaviors were evaluated using object location recognition (OLR) experiment, novel object recognition (NOR) test, Morris water maze (MWM) test and passive avoidance (PA) task. Cholinergic system function and oxidative stress activities were measured in the hippocampus. The expression levels of CREB, Egr-1, c-Fos and c-Jun were also measured in the hippocampus. Results: Rh1 administration could improve the cognitive performance of scopolaminetreated mice in the object location recognition, the novel object recognition, the Morris water maze and the passive avoidance tests. And Rh1 significantly enhanced cholinergic system function and suppressed oxidative stress level in the hippocampus of scopolaminetreated mice, as indicated by decreasing acetylcholinesterase (AChE) activity, elevating choline acetyltransferase (ChAT) activity and acetylcholine (Ach) level, increasing superoxide dismutase (SOD) activity and lowering the level of malondialdehyde (MDA) in the hippocampus of scopolamine-treated mice. Moreover, for the first time, Rh1 was found to significantly upregulate the expression levels of CREB, early growth response-1 (Egr-1), c-Fos and c-Jun in the hippocampus of scopolamine-treated mice. Conclusion: These results demonstrated that Ginsenoside Rh1 might exert a significant neuroprotective effect on cognitive dysfunctions induced by scopolamine, driven in part by the modulation of cholinergic activity, oxidative stress level and upregulation of CREB, Egr-1, c-Fos and c-Jun expression in the hippocampus.

Surface-engineered particulate delivery systems for vaccine administration have been widely investigated in experimental and clinical studies. However, little is known about charge-coated microspheres as potential recombinant subunit protein antigen delivery systems in terms of adsorption and related immune responses. In the present study, cationic polymers, including chitosan (CS), chitosan chloride (CSC), and polyethylenimine (PEI), were used to coat PIA microspheres to build positively charged surfaces. Antigen adsorption capacity was enhanced with increased surface charge of coated microspheres. In macrophages, HBsAg adsorbed on the surface of cationic microspheres specifically enhanced antigen uptake and augmented CD86, MHC I, and MHC II expression and IL-1 beta, IL-6, TNF-alpha, and IL-12 release. Antigens were more likely to localize independent of lysosomes after phagocytosis in antigen-attached cationic microsphere formulations. After intraperitoneal immunization, cationic microsphere-based vaccine formulations generated a rapid and efficient humoral immune response and cytokine release as compared with aluminum-adsorbed vaccine and free antigens in vivo. Moreover, microspheres coated with cationic polymers with relatively high positive charges and higher antigen adsorption exhibited strong stimulation of the Th1 response. In conclusion, PLA microspheres coated with cationic polymers may be a potential recombinant antigen delivery system to induce strong cell and humoral immune responses.

Ethnopharmacological relevance Dragon's blood (Chinese name: Xuejie), which comprises red resins obtained from several plants (27 species from 4 families), is drawing worldwide interests in medicinal applications owing to its broad pharmacological spectrum such as promoting blood circulation, regenerating muscle, relieving swelling and pain, maintaining hemostasis, etc. Aim of the study This work aims to evaluate current research progress on phenolic constituents, pharmacological activities, quality control, and metabolism of six Dracaena plants, namely, Dracaena cochinchinensis (Lour.) S.C.Chen, D. cambodiana Pierre ex Gagnep., D. cinnabari Balf. f., D. draco (L.) L., D. loureiroi Gagnep., and D. schizantha Baker, figure out the shortcomings of existing studies, and provide meaningful guidelines for future investigations. Methods Extensive database retrieval, such as SciFinder, PubMed, CNKI, ChemSpider, etc., was performed by using the keywords “Dracaena,” “dragon's blood,” as well as the Latin names of the six Dracaena species. In addition, relevant textbooks, patents, reviews, and documents were also employed to ensure sufficient information is collected. Results Flavonoids and their oligomers are the primary chemical clusters distributed in Dracaena plants. Pharmacological activities including analgesic, anti-inflammatory, antibacterial, hypolipidemic, hypoglycemic, and cytotoxic effects; bi-directional regulation effects on hemorheology; and cardiovascular and cerebrovascular effects have been disclosed by modern pharmacological evaluations. The chemical and metabolic profiles after oral administration of dragon's blood extract were preliminarily characterized. However, some of the pharmacological investigations reported only elementary methodologies and unreliable findings, and even worse, some important aspects were questionable or missing in these articles. Conclusions Dragon's blood is a valuable source of bioactive compounds, mainly flavonoids and their oligomers. Its potential therapeutic effects on different diseases are attractive, such as the notable effect on cardiovascular diseases. In future studies, there is an urgent need to test the effect of this extract on appropriate cell lines and animal models to analyze its ethnopharmacological applications; moreover, “composition−effect correlation” methods and omics technologies are demanded for identifying the effective material basis and therapeutic mechanisms before entering into clinical trials. Moreover, attention should be paid to the chemical profiling and quality evaluation of this precious herbal medicine.

Effects of salicylic acid (SA) on gibberellin (GA) homeostasis, C-repeat/dehydration-responsive element binding factor (CBF) pathway, and antioxidant enzyme systems linked to chilling- and oxidative-stress tolerance in tomato fruit were investigated. Mature green tomatoes (Solanum lycopersicum L. cv. Moneymaker) were treated with 0, 0.5, and 1 mM SA solution for 15 min before storage at 4 °C for 28 days. In comparison to 0 or 0.5 mM SA, 1 mM SA significantly decreased the chilling injury (CI) index in tomato fruit. In the SA-treated fruit, the upregulation of GA biosynthetic gene (GA3ox1) expression was followed by gibberellic acid (GA3) surge and DELLA protein degradation. CBF1 participated in the SA-modulated tolerance and stimulated the expression of GA catabolic gene (GA2ox1). Furthermore, 1 mM SA enhanced activities of antioxidant enzymes and, thus, reduced reactive oxygen species accumulation. Our findings suggest that SA might protect tomato fruit from CI and oxidative damage through regulating GA ...

20(S)-protopanaxadiol (PPD) possesses various biological properties, including anti-inflammatory, antitumor and anti-fatigue properties. Recent studies found that PPD functioned as a neurotrophic agent to ameliorate the sensory deficit caused by glutamate-induced excitotoxicity through its antioxidant effects and exhibited strong antidepressant-like effects in vivo. The objective of the present study was first to investigate the effect of PPD in scopolamine (SCOP)-induced memory deficit in mice and the potential mechanisms involved. In this study, mice were pretreated with PPD (20 and 40 μmol/kg) and donepezil (1.6 mg/kg) intraperitoneally (i.p) for 14 days. Then, open field test was used to assess the effect of PPD on the locomotor activity and mice were daily injected with SCOP (0.75 mg/kg) to induce cognitive deficits and then subjected to behavioral tests by object location recognition (OLR) experiment and Morris water maze (MWM) task. The cholinergic system function, oxidative stress biomarkers and protein expression of Egr-1, c-Fos, and c-Jun in mouse hippocampus were examined. PPD was found to significantly improve the performance of amnesia mice in OLR and MWM tests. PPD regulated cholinergic function by inhibiting SCOP-induced elevation of acetylcholinesterase (AChE) activity, decline of choline acetyltransferase (ChAT) activity and decrease of acetylcholine (Ach) level. PPD suppressed oxidative stress by increasing activities of antioxidant enzymes such as superoxide dismutase (SOD) and lowering maleic diadehyde (MDA) level. Additionally, PPD significantly elevated the expression of Egr-1, c-Fos, and c-Jun in hippocampus at protein level. Taken together, all these results suggested that 20(S)-protopanaxadiol (PPD) may be a candidate compound for the prevention against memory loss in some neurodegenerative diseases such as Alzheimer's disease (AD).

Therapeutic vaccination for the treatment of chronic hepatitis B is promising but has so far shown limited clinical efficacy. Herein, we employ polylactide nanoparticles (NPs) as the vaccine adjuvant and systematically explore their effect on activation of specific immunity and the underlying theoretical mechanisms. In vitro studies show that hepatitis B surface antigen (HBsAg) accumulates in antigen-presenting cells (APCs) to a larger content (270%) with the assistant of NP in comparison with the pure-antigen group. Besides the elevated costimulators (CD80/86) and increased major histocompatibility complex (MHC) II molecules, the MHC I molecules are also found upregulated. This result is mostly owing to the divergent antigen trafficking ways of NP-antigen in APCs, especially for the escape of exogenous HBsAg from the lysosomes to the cytosol. Interestingly, the MHC I level is downregulated in alum-antigen group, indicating a possible reason for its inefficiency in priming cellular response. Further in vivo experiments establish that NP-antigen group indeed enhances the CD8(+) CTL cytotoxicity and IFN-gamma cytokine secretion. Meanwhile, specific antibody titer is also upregulated, and even surpasses that of the commercialized alum-antigen. All these results strongly support that NP-based antigen promotes an orchestration of cellular and humoral immune response, exhibiting favorable intrinsic properties to be applied in therapeutic vaccines. (C) 2011 Elsevier Ltd. All rights reserved.

Nitric oxide (NO), an important signaling molecule in plants, is known to play crucial roles in the regulation of both defense responses and inducing resistance to fungal pathogens. To investigate NO synthesis during induced resistance and its mechanism, green mature tomatoes ( Lycopersicon esculentum cv. No.4 Zhongshu) were treated with the elicitor from Botrytis cinerea , considered an effective inducer of disease resistance, nitric oxide synthase (NOS) inhibitor N-nitro- l -arginine ( l -NNA) and nitric reductase (NR) inhibitor sodium azide (NaN 3 ). Our data indicated that reduced disease symptoms, and lesion size, in tomato fruit with B. cinerea were inhibited by 93.3 and 80.9% respectively three and four days after inoculation. NO biosynthesis was rapidly initiated one day after green mature tomatoes were treated with the elicitor and the peak NO content reached 3.43 μmol/g FW. The activities of NOS and defense enzymes, including phenylalanine ammonia-lyase (PAL), chitinase (CHI), β-1,3-glucanase (GLU), and polyphenoloxidase (PPO), were also increased by the elicitor treatment. Gene expression of pathogenesis-related protein 1(PR-1) increased within 2 h, and total phenols contents increased significantly within four days. A treatment of NOS inhibitor + elicitor increased disease incidence and lesion area and reduced the activities of defense enzymes compared with elicitor-treated fruit, indicating that elicitor-induced NO generation and disease resistance could be blocked by a NOS inhibitor. The NR inhibitor did not obviously influence the content of NO and disease resistance after incubation with the elicitor, and elicitor treatment did not induce NR activity. These results imply that NOS is a key enzyme regulating NO production in induced resistance.

Methyl jasmonate (MeJA), a major derivative of the plant hormone jasmonic acid, plays a critical role in inducing resistance to fungal pathogen. To study the endurance of MeJA-induced resistance and its cause, green mature tomatoes (Solanum esculentum cv. Lichun) were treated with 100 μM MeJA and nordihydroguaiaretic acid (NDGA, LOX inhibitor) at −35 kPa for 0.5 min and incubated at 25 ± 1 °C, 85–90% RH. Treatment with MeJA reduced disease symptoms in tomato fruit soon after being inoculated with Botrytis cinerea. Lesion size in MeJA-treated fruit was inhibited by 42.5%, 27.9% and 13.9% respectively (P < 0.05) in fruit inoculated 1, 3 and 6 d after treatments. At advanced stages (inoculation carried out 9 and 12 d after treatments), no inhibitory effect of MeJA were found. Ethylene biosynthesis was activated in the response of green mature tomatoes to methyl jasmonate with a rapid (1 d) and enhanced ethylene peak (0.9 ng kg−1 FW s−1). However the ethylene level was below that of the control from 6 d to 12 d. This rise was closely related with conversion of ACC to ethylene, especially a rise in ACO activity (6 h), which preceded an increase in ACS (12 h) after MeJA treatment. The development of ethylene biosynthesis was accompanied by a significant increase in LOX activity. Two significant O2− peaks (P < 0.05) were detected in MeJA-treated fruit during storage (6.18 μmol g−1 FW min−1 at 6 h and 5.68 μmol g−1 FW min−1 at 3 d). The correlations between LOX, and O2− and ACO activities were 0.75, 0.73 respectively (P < 0.05). These results indicate that MeJA-induced resistance against B. cinerea is durable, MeJA induces LOX and the superoxide radicals formed by LOX may activate ACO and ethylene biosynthesis.

BACKGROUND: Nitric oxide (NO) and hydrogen peroxide (H2O2) are signal molecules involved in the disease response of plants, and there is a close relationship between them. To investigate the interaction of NO and H2O2 during disease resistance response in postharvest fruits and vegetables, tomato (Lycopersicon esculentum cv. Lichun) fruits were treated after harvest by vacuum infiltration with NO donor and H2O2 scavenger. RESULTS: The resistance of tomato fruits to Rhizopus nigricans Ehrenb. invasion and the activities of defensive enzymes phenylalanine ammonia-lyase, chitinase and glutathione S-transferase were enhanced by NO. However, these effects of NO on resistance were weakened by H2O2 scavenger, which showed that H2O2 was required in NO-mediated disease resistance in harvested tomato. Meanwhile, the endogenous H2O2 level was dual-regulated by NO. During the earlier period of storage the H2O2 peak was advanced by NO, but during the later period of storage, when the concentration of H2O2 was relatively higher, H2O2 accumulation was delayed by NO. The activities of superoxide dismutase, ascorbate peroxidase and catalase were involved in this regulation. CONCLUSION: The results imply that application of exogenous NO can enhance the disease resistance of postharvest tomato; NO may interact with H2O2 and exert its effect by modulating the endogenous H2O2 level. This result is useful for the further investigation and application of NO in postharvest disease control. Copyright © 2008 Society of Chemical Industry

The present study evaluated the significance of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in 30 patients with endometrial carcinoma and the relationship of those molecular markers to tumor characteristics and microvessel density (MVD). Immunohistochemical expression of COX-2, iNOS, and CD34 was analyzed on paraffin-embedded tissue sections. The COX-2 and iNOS positive rates were 66.7% and 73.3%, respectively. The level of COX-2 expression was higher in grade II tumors than in grade III tumors (p0.05). The percentage of iNOS positivity was higher in patients with deep myometrial invasion than in patients without or less than 50% myometrial invasion (p0.05). There was significant correlation between positive COX-2 and positive iNOS expression (r = 0.601, p0.001). Both COX-2 and iNOS were significantly correlated with MVD (r = 0.02 p0.05; r = 0.599 p0.0001, respectively). The present findings suggest that combined expression of COX-2 and iNOS may play an important role in development and invasion of endometrial cancer and that this could be partially attributable to modulation of angiogenesis by COX-2 and iNOS.

Human plasma fraction IV is an intermediate precipitate during the production of human serum albumin using cold ethanol method. Haptoglobin locates in this fraction can be purified for various applications. A new process integration of polyethylene glycol (PEG) precipitation and ion-exchange chromatography (IEC) was developed for purification of haptoglobin, which could effectively purify the haptoglobin from 16.6% to 95%. The recovery of the new process was 58.2% in comparison to 30.3% of the conventional affinity chromatography. Furthermore, 175 mg haptoglobin production in a scaled-up process showed the method to be simple, fast, and low-cost.

Metallothioneins (MTs), as a family of low-molecular-weight, cysteine-rich, and metal-binding proteins, show potential for utilization in functional food. Tomato plants were transformed with gene constructs that contained mt-I encoding the mouse MT-I, similar in sense orientation with the constitutively active double 35S promoter from cauliflower mosaic virus. Three independent transformants, which had copies of the gene in their genomes, were obtained. In these transgenic lines, high-level expression of MT-I, high zinc content, and some antioxidant enzyme activities were detected in leaves. The average zinc content in transgenic tomato leaves was 32.7 mg/100 g FW, which about 1.6 times higher than that in wild-type. The superoxide dismutase activity was also higher (68.6, 66.9, and 66.1 U/g FW in the three transformants) than that in wild-type (57.4 U/g FW). In particular, the levels of superoxide free radical scanvenging in the three transformants were 14.2%, 14.6%, and 13.7%, respectively, which about 1.5 times higher than that in control (5.6%). Transgenic MT tomato may potentially be used as an antioxidant and for zinc supplementation.

Polyaniline nanomaterial (nPANI) is getting popular in many industrial fields due to its conductivity and stability. The fate and effect of nPANI in the environment is of paramount importance towards its technological applications. In this work, the cytotoxicity of nPANI, which was prepared by rapid surface polymerization, was studied on rat celiac macrophages. Cell viability of macrophages treated with various concentrations of nPANI and different periods ranging from 24 to 72 hours was tested by a MTT assay. Damages of nPANI to structures of macrophages were evaluated according to the exposure level of cellular reactive oxygen species (ROS) and change of mitochondrial membrane potential (MMP). We observed no significant effects of nPANI on the survival, ROS level and MMP loss of macrophages at concentrations up to 1 µg/ml. However, higher dose of nPANI (10 µg/ml or above) induced cell death, changes of ROS level and MMP. In addition, an increase in the expression level of caspase-3 protein and its activated form was detected in a Western blot assay under the high dose exposure of nPANI. All together, our experimental results suggest that the hazardous potential of nPANI on macrophages is time- and dose-dependent and high dose of nPANI can induce cell apoptosis through caspase-3 mediated pathway.

C-Repeat/dehydration-responsive element binding factor (CBF) is a transcription factor regulating cold response in plants, of which little is known in fruits. We showed a double-peak expression pattern of Lycopersicon esculentum putative transcriptional activator CBF1 (LeCBF1) in mature green fruit. The peaks appeared at 2 and 16 h after subjection to cold storage (2 °C). The second peak was coincident with, and thus caused by a peak in endogenous ethylene production. We showed that LeCBF1 expression was regulated by exogenous ethylene and 1-methylcyclopropene, and was not expressed without cold induction. LeCBF1 expression was different in the five maturation stages of fruits, but expression peaked at 2 h at all stages.