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The vacuolar proton-pumping pyrophosphatase gene (VPP) is often used to enhance plant drought tolerance via genetic engineering. In this study, the drought tolerance of four transgenic inbred maize lines overexpressing ZmVPP1 (PH4CV-T, PH6WC-T, Chang7-2-T, and Zheng58-T) and their transgenic hybrids was evaluated at various stages. Under normal and drought conditions, the height and fresh weight were greater for the four transgenic inbred maize lines than for the wild-type (WT) controls at the germination and seedling stages. Additionally, the transgenic plants exhibited enhanced photosynthetic efficiency at the seedling stage. In irrigated and non-irrigated fields, the four transgenic lines grew normally, but with increased ear weight and yield compared with the WT plants. Moreover, the ear weight and yield of the transgenic hybrids resulting from the PH4CV-T×PH6WC-W and Chang7-2-T×Zheng58-W crosses increased in the non-irrigated field. Our results demonstrated that the growth and drought tolerance of four transgenic inbred maize lines with improved photosynthesis were enhanced by the overexpression of ZmVPP1. Moreover, the Chang7-2 and PH4CV transgenic lines may be useful for future genetic improvements of maize hybrids to increase drought tolerance.

Purpose Moesin-ezrin-radixin-like protein (Merlin) has been identified as a tumor suppressor in several types of cancers. However, the biological function of Merlin in osteosarcoma remains unclear. MicroRNAs (miRNAs) can influence cancer progression by targeting oncogenes or anti-oncogenes. In this study, we sought to evaluate the regulation of Merlin expression by miR-25-3p and the role of the miR-25-3p/Merlin axis in osteosarcoma progression, with the aim of identifying a potential therapeutic target for osteosarcoma. Materials and methods TCGA (The Cancer Genome Atlas) database was used to analyze the correlation between Merlin expression and prognosis. RT-qPCR and Western blotting analyses were performed to compare Merlin expression between normal and malignant cells. A dual-luciferase reporter assay was performed to evaluate the direct targeting of Merlin by miR-25-3p. We overexpressed miR-25-3p, or/and Merlin, in U-2 OS and 143B cells, and studied their cellular functions in vitro. MTT and colony formation assays were performed to determine the effects on cell growth. EdU and cell cycle assays were performed to analyze the effects in cell replication. We used annexin V-fluorescein isothiocyanate and propidium iodide to stain apoptotic cells, and analyzed the cells using flow cytometry. The effects on cell metastasis were studied in wound healing and transwell assays. Lastly, the underlying mechanism was determined in RT-qPCR and Western blotting experiments. Results Low Merlin expression was linked to poor prognosis. miR-25-3p was observed to directly target Merlin and downregulate its expression. miR-25-3p promoted cell growth, migration, and invasion, and inhibited apoptosis induced by cisplatin. Moreover, the overexpression of Merlin reversed the abovementioned effects of miR-25-3p. Further, the miR-25-3p/Merlin axis was observed to play an important role in the Hippo pathway, and regulated the expression of genes such as BIRC5, CTGF, and CYR61. Conclusion miR-25-3p functions as an oncogenic microRNA in osteosarcoma by targeting Merlin, and may serve as a potential therapeutic target for osteosarcoma.

Cytochrome P450 monooxygenases (CYPs) play essential physiological functions in insects. CYP303A1 is highly conserved in insect species studied to date, and shows an indispensable role for adult eclosion in both Locusta migratoria and Drosophila melanogaster. However, how CYP303A1 is regulated to control insect developmental process remains uninvestigated. In this study, we discovered functional binding sites for miR-184 in the coding sequence of LmCYP303A1. The luciferase reporter assay showed that miR-184 could target LmCYP303A1 and regulate its expression in vitro. Furthermore, overexpression of miR-184 through microinjection of agomir to locusts reduced the transcripts of LmCYP303A1 and led to the abnormal molting, which is similar to the phenotype of silencing LmCYP303A1 by direct injection of dsLmCYP303A1 to locusts. Meanwhile, down regulation of miR-184 by injection of antagomir increased the LmCYP303A1 transcript and caused molting defects. These findings suggested that miR-184 could target LmCYP303A1 to regulate the molting process in L. migratoria, which might be considered as a novel target for pest control. This article is protected by copyright. All rights reserved.

Objective To explore the possible preventive mechanism of Hunan expert group recommended Chinese medicine prescription of No. 2 (Pre-No. 2) against coronavirus disease 2019 (COVID-19) by network pharmacology method. Methods The target proteins of effective components and active compounds in Pre-No. 2 were screened by searching the Tradi-tional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). A component-target-disease interac-tion network of Pre-No. 2 was constructed by Cytoscape 3.7.2, gene ontology (GO) analysis, and Kyoto encyclopedia of genes and genomes (KEGG) analysis of target protein pathway by DAVID. Results A total of 163 compounds and 278 target protein targets in Pre-No. 2 were collected from the TCMSP database. Kaempferol, wogonin, 7-methoxy-2-methyl isoflavone, formononetin, isorhamnetin, and licochalcone A were the most frequent targets in the regulatory network. GO enrichment analysis showed that Pre-No. 2 regulated response to virus, viral processes, humoral immune responses, defense responses to virus and viral entry into host cells. KEGG enrichment analysis showed that the formula regulated the NF-κB signaling pathway, B cell receptor signaling pathway, viral carcinogenesis, T cell signaling pathway and FcγR-mediated phagocytosis signaling pathway. Conclusions Pre-No. 2 may play a preventive role against COVID-19 through regulation of the Toll-like signaling, T cell signaling, B cell signaling and other signaling pathways. It may re-gulate the immune system to protect against anti-influenza virus.

Antibiotic-resistant infections are predicted to kill 10 million people worldwide per year by 2050 and to cost the global economy 100 trillion USD. Novel approaches and alternatives to conventional antibiotics are urgently required to combat antimicrobial resistance. We have synthesized a chitosan-based oligolysine antimicrobial peptide, CSM5-K5 (where CSM denotes chitosan monomer repeat units and K denotes lysine amino acid repeat units), that targets multidrug-resistant (MDR) bacterial species. Here, we show that CSM5-K5 exhibits rapid bactericidal activity against methicillin-resistant

Recently, we read with great interest the article published in Cell by Yu et al, which reported that TAR DNA-binding protein 43 (TDP-43) overexpression stimulated the released of mitochondrial DNA (mtDNA) into the cytoplasm through mitochondrial permeability transition pore. Then, mtDNA bound to cyclic GMP-AMP synthase (cGAS) to activate the cGAS/STING pathway, which caused the inflammation of amyotrophic lateral sclerosis (ALS).

Plant height (PH) and ear height (EH) are important factors for maize plant type and grain yield. Weighted gene co-expression network analysis (WGCNA) is an important method to explain the relationships between gene network and complicated traits and identify the PH and EH associated genes. In this study, we used Zheng 58, Ye 478, Chang 7-2, Huangzaosi and its combinations Zhengdan 958, Anyu 5, Zheng 58/Huangzaosi, and Ye 478/Huangzaosi as materials and utilized transcriptome data under the planting densities of 45,000 plants hm–2 and 67,500 plants hm–2 to construct a co-expression network by WGCNA, getting 24 and 21 co-expression modules, respectively. Among them, a total of 15 co-expression modules were significantly correlated with PH and EH, with the absolute correlation coefficients higher than 0.50. Six modules were overlapped between PH and EH. By gene function analysis, these overlapped modules were significantly enriched in development, photosynthesis, response to light stimulus, plant hormone, and carbohydrate biosynthesis/metabolism related activities. According to connectivity of genes in modules, AP2-EREBP transcription factor EREB14 , thiaminase TENA2 , phosphoglyceric kinase PGK , glutathione transferase GST2 , and succinate dehydrogenase SUDH7 were considered as hub genes. From gene networks, EREB14 was connected with three known PH genes D8 , DWF1 , ZmGRF10 , and C3H35 (C3H transcription factor), GATA4 (C2C2-GATA transcription factor), and ethylene homology ETR40 . Reported PH genes An1 and GA20ox3 were also found in our co-expression modules. From the networks of the five known PH genes, ARF-transcription factor 7 ( ARFTF7 ), ARFTF26 , GST39 , photosystem II oxygen evolving polypeptide PspB2 , and photosystem I N subunit PasN1 had connections with these known PH genes. The identification of 15 co-expression modules and their hub genes, and analysis of their gene function and gene networks of key genes will be helpful for revealing the genetic basis of PH and EH.

Major infectious diseases, such as viral hepatitis, acquired immune deficiency syndrome, and tuberculosis, are difficult to eliminate within a short period. Dengue fever, global influenza pandemic, and ebola and other high-threat pathogens are now the main threats to human health as well. Therefore, it is particularly important to find new strategies for the prevention and treatment of infectious diseases. The human gut tract contains trillions of microbial cells, including bacteria, archaea, fungi and viruses. These microbes are defined as gut microbiota and play an important role in human development, immunity, metabolism and diseases. In this review, we analyzed the mechanisms by which alterations in gut microbiota affect infectious diseases and how infectious diseases regulate the structure and function of gut microbiota. Finally, we summarized and discussed methods used for the diagnosis, prevention and treatment of infectious diseases based on the alteration of gut microbiota.

A worth noticing pattern in current invasive biology is the clonal ability of many of the world's worst invasive plants. Selective placement of ramets (i.e. foraging behavior) can intensify ramet performance and allocation, and place more ramets in the more favorable microhabitats, which can maximum utilize resource and share risk in heterogeneous environments. Still little is known about whether invasive alien and native clonal plants differ in the selective placement patterns of ramets in invasive clonal plants or not. We used five congeneric pairs of naturally co-occurring invasive alien and native clonal plant species in China. In a glasshouse, we grew all species in pots under a homogeneous and three heterogeneous conditions (i.e. light, soil nutrients or water) subjected to resource-high or -low patches. All biomass parameters and number of ramets significantly increased in resource-high patches in all three types of heterogeneous environments. Interestingly, growth of invasive alien plants benefited significantly more from resource-high patches than native plants in all heterogeneous environments. Overall, invasive had higher biomass parameters per ramet than natives. Ramet parameters of invasive plants also benefited more from resource-low patches than natives. Three different selective placement patterns of ramets in resource-low patches were exhibited in invasive plants: ramet increasing shoot investment (above pattern), increasing root investment (below pattern) and increasing both investments (complete pattern) in the light, soil water and nutrient heterogeneity, respectively. Investment on less, larger ramet was the adaptive strategy of invasive plants in resource-poor patches. The results suggest that adaptively selective placement patterns of ramets promote a higher morphology plasticity and performance in invasive clonal plants over natives. When alien clonal plants spread new areas with light, soil nutrients or water heterogeneity, selective placement patterns of ramets might play an important role in plant performance and competitive superior by capitalizing more on additional resources.

Background and aims: Studies of hereditary transthyretin amyloidosis (ATTRv amyloidosis) in South-East Asia are underrepresented in the literature. We report the unique phenotypic and genetic characteristics of this disorder in a multiracial South-East Asian cohort. Methods: Patients with genetically proven ATTRv amyloidosis were identified over a 13-year period (2007–2020) at the National Neuroscience Institute, Singapore. Clinical, laboratory, genotypic and electrophysiological features were retrospectively reviewed. Results: 29 patients comprising Chinese, Malay, Burmese, Vietnamese and Indonesians with ATTRv amyloidosis were identified. Somatic neuropathy was the most common initial presentation, followed by carpal tunnel syndrome, autonomic dysfunction and cardiac dysfunction. ATTR-A97S (p.Ala117Ser) was the most common variant found in 14 patients, constituting 66.7%of ethnic Chinese patients and 48.3%of the entire cohort. Five patients had early-onset disease (age

Nanofibers can mimic natural tissue structure by creating a more suitable environment for cells to grow, prompting a wide application of nanofiber materials. In this review, we include relevant studies and characterize the effect of nanofibers on mesenchymal stem cells, as well as factors that affect cell adhesion and osteogenic differentiation. We hypothesize that the process of bone regeneration in vitro is similar to bone formation and healing in vivo, and the closer nanofibers or nanofibrous scaffolds are to natural bone tissue, the better the bone regeneration process will be. In general, cells cultured on nanofibers have a similar gene expression pattern and osteogenic behavior as cells induced by osteogenic supplements in vitro. Genes involved in cell adhesion (focal adhesion kinase (FAK)), cytoskeletal organization, and osteogenic pathways (transforming growth factor-β (TGF-β)/bone morphogenic protein (BMP), mitogen-activated protein kinase (MAPK), and Wnt) are upregulated successively. Cell adhesion and osteogenesis may be influenced by several factors. Nanofibers possess certain physical properties including favorable hydrophilicity, porosity, and swelling properties that promote cell adhesion and growth. Moreover, nanofiber stiffness plays a vital role in cell fate, as cell recruitment for osteogenesis tends to be better on stiffer scaffolds, with associated signaling pathways of integrin and Yes-associated protein (YAP)/transcriptional co-activator with PDZ-binding motif (TAZ). Also, hierarchically aligned nanofibers, as well as their combination with functional additives (growth factors, HA particles, etc.), contribute to osteogenesis and bone regeneration. In summary, previous studies have indicated that upon sensing the stiffness of the nanofibrous environment as well as its other characteristics, stem cells change their shape and tension accordingly, regulating downstream pathways followed by adhesion to nanofibers to contribute to osteogenesis. However, additional experiments are needed to identify major signaling pathways in the bone regeneration process, and also to fully investigate its supportive role in fabricating or designing the optimum tissue-mimicking nanofibrous scaffolds.

Characterisation of protein function based solely on homology searches may overlook functions under specific environmental conditions, or the possibility of a protein having multiple roles. In this study we investigated the role of YtfB, a protein originally identified in a genome-wide screen to cause inhibition of cell division, and has demonstrated to localise to the Escherichia coli division site with some degree of glycan specificity. Interestingly, YtfB also shows homology to the virulence factor OapA from Haemophilus influenzae, which is important for adherence to epithelial cells, indicating the potential of additional function(s) for YtfB. Here we show that E. coli YtfB binds to N’acetylglucosamine and mannobiose glycans with high affinity. The loss of ytfB results in a reduction in the ability of the uropathogenic E. coli strain UTI89 to adhere to human kidney cells, but not to bladder cells, suggesting a specific role in the initial adherence stage of ascending urinary tract infections. Taken together, our results suggest a role for YtfB in adhesion to specific eukaryotic cells, which may be additional, or complementary, to its role in cell division. This study highlights the importance of understanding the possible multiple functions of proteins based on homology, which may be specific to different environmental conditions.

Metabolic reprogramming has become a hot topic recently in the regulation of tumour biology. Although hundreds of altered metabolic genes have been reported to be associated with tumour development and progression, the important prognostic role of these metabolic genes remains unknown. We downloaded messenger RNA expression profiles and clinicopathological data from The Cancer Genome Atlas and the Gene Expression Omnibus database to uncover the prognostic role of these metabolic genes. Univariate Cox regression analysis and lasso Cox regression model were utilized in this study to screen prognostic associated metabolic genes. Patients with high-risk demonstrated significantly poorer survival outcomes than patients with low-risk in the TCGA database. Also, patients with high-risk still showed significantly poorer survival outcomes than patients with low-risk in the GEO database. What is more, gene set enrichment analyses were performed in this study to uncover significantly enriched GO terms and pathways in order to help identify potential underlying mechanisms. Our study identified some survival-related metabolic genes for rectal cancer prognosis prediction. These genes might play essential roles in the regulation of metabolic microenvironment and in providing significant potential biomarkers in metabolic treatment.

Maximizing seed yield is the ultimate breeding goal in important cereal crop species. Seed set is a key developmental stage in the process of seed formation, which determines grain number, seed mass, and realized yield potential, and can be severely affected by abiotic and biotic stresses. However, seed set can also be substantially reduced by genetic factors even under optimal fertilization conditions. The underlying molecular genetic mechanisms are still obscure. In this review, we elucidate the process of seed set of cereal crop species in detail, including development of floral structures, formation of viable gametes, double fertilization, seed development, and abortion. We discuss how genetic and non-genetic factors affect seed set in different development stages. Finally, we will propose novel strategies to study genetic mechanisms controlling seed set and exploit genetic resources to improve seed set in cereal crop species.

Spinal cord injury (SCI) normally results in cell death, scarring, cavitation, inhibitory molecules release, etc., which are regarded as a huge obstacle to reconnect the injured neuronal circuits because of the lack of effective stimulus. In this study, a functional gelatin sponge scaffold was used to inhibit local inflammation, enhance nerve fiber regeneration, and improve neural conduction in the canine. This scaffold had good porosity and modified with neurotrophin-3 (NT-3)/fibroin complex, which showed sustained release in vitro. After the scaffold was transplanted into canine spinal cord hemisection model, hindlimb movement, and neural conduction were improved evidently. Migrating host cells, newly formed neurons with associated synaptic structures together with functional blood vessels with intact endothelium in the regenerating tissue were identified. Taken together, the results demonstrated that using bioactive scaffold could establish effective microenvironment stimuli for endogenous regeneration, providing a potential and practical strategy for treatment of spinal cord injury. © 2018 The Authors Journal of Biomedical Materials Research Part A Published by Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2158-2170, 2018.

Enterococcus faecalis and Enterococcus faecium are common inhabitants of the human gastrointestinal tract, as well as frequent opportunistic pathogens. Enterococci cause a range of infections including, most frequently, infections of the urinary tract, catheterized urinary tract, bloodstream, wounds and surgical sites, and heart valves in endocarditis. Enterococcal infections are often biofilm-associated, polymicrobial in nature, and resistant to antibiotics of last resort. Understanding Enterococcal mechanisms of colonization and pathogenesis are important for identifying new ways to manage and intervene with these infections. We review vertebrate and invertebrate model systems applied to study the most common E. faecalis and E. faecium infections, with emphasis on recent findings examining Enterococcal-host interactions using these models. We discuss strengths and shortcomings of each model, propose future animal models not yet applied to study mono- and polymicrobial infections involving E. faecalis and E. faecium, and comment on the significance of anti-virulence strategies derived from a fundamental understanding of host-pathogen interactions in model systems. NRF (Natl Research Foundation, S’pore) MOE (Min. of Education, S’pore) Published version

Background An impairment of the integrity of the mucosal epithelial barrier can be observed in the course of various gastrointestinal diseases. The migration and proliferation of the intestinal epithelial (IEC-6) cells are essential repair modalities to the healing of mucosal ulcers and wounds. Atractylenolide I (AT-I), one of the major bioactive components in the rhizome of Atractylodes macrocephala Koidz. (AMR), possesses multiple pharmacological activities. This study was designed to investigate the therapeutic effects and the underlying molecular mechanisms of AT-I on gastrointestinal mucosal injury. Methods Scratch method with a gel-loading microtip was used to detect IEC-6 cell migration. The real-time cell analyzer (RTCA) system was adopted to evaluate IEC-6 cell proliferation. Intracellular polyamines content was determined using high performance liquid chromatography (HPLC). Flow cytometry was used to measure cytosolic free Ca2+ concentration ([Ca2+]c). mRNA and protein expression of TRPC1 and PLC-γ1 were determined by real-time PCR and Western blotting assay respectively. Results Treatment of IEC-6 cells with AT-I promoted cell migration and proliferation, increased polyamines content, raised cytosolic free Ca2+ concentration ([Ca2+]c), and enhanced TRPC1 and PLC-γ1 mRNA and protein expression. Depletion of cellular polyamines by DL-a-difluoromethylornithine (DFMO, an inhibitor of polyamine synthesis) suppressed cell migration and proliferation, decreased polyamines content, and reduced [Ca2+]c, which was paralleled by a decrease in TRPC1 and PLC-γ1 mRNA and protein expression in IEC-6 cells. AT-I reversed the effects of DFMO on polyamines content, [Ca2+]c, TRPC1 and PLC-γ1 mRNA and protein expression, and restored IEC-6 cell migration and proliferation to near normal levels. Conclusion Our data demonstrate that AT-I stimulates intestinal epithelial cell migration and proliferation via the polyamine-mediated Ca2+ signaling pathway. Therefore, AT-I may have the potential to be further developed as a promising therapeutic agent to treat diseases associated with gastrointestinal mucosal injury, such as inflammatory bowel disease and peptic ulcer.

Antibiotic-resistant infections are predicted to kill 10 million people worldwide per year by 2050 and to cost the global economy 100 trillion USD. Novel approaches and alternatives to conventional antibiotics are urgently required to combat antimicrobial resistance. We have synthesized a chitosan-based oligolysine antimicrobial peptide, CSM5-K5, which targets multidrug resistant (MDR) bacterial species. Here we show that CSM5-K5 exhibits rapid bactericidal activity against methicillin resistantStaphylococcus aureus(MRSA), MDREscherichia coli, and vancomycin resistantEnterococcus faecalis(VRE). Combinatorial therapy of CSM5-K5 with antibiotics to which each organism is otherwise resistant restores sensitivity to the conventional antibiotic. CSM5-K5 alone significantly reduced pre-formed bacterial biofilm by two-four orders of magnitude and, in combination with traditional antibiotics, reduced pre-formed biofilm by more than two-three orders of magnitude at sub inhibitory concentrations. Moreover, using a mouse excisional wound infection model, CSM5-K5 treatment reduced bacterial burdens by one to three orders of magnitude, and acted synergistically with vancomycin and tetracycline to clear VRE and MDRE. coli,respectively. Importantly, little to no resistance against CSM5-K5 arose for any of the three MDR bacteria during 15 days of serial passage. This work demonstrates the feasibility and benefits of using this synthetic cationic peptide as alternative to, or in combination with, traditional antibiotics to treat infections caused by MDR bacteria.

Enterococcus faecalis is one of the leading causes of hospital-acquired infections (HAIs). One of the most commonly associated virulence factors in these HAIs is the endocarditis and biofilm-associated pilus (Ebp) which is often implicated in biofilm formation. E. faecalis is associated with urinary tract infections (UTI), bacteremia, endocarditis, surgical wounds, and many other infections. These infections likely give rise to unfolded or misfolded proteins, leading to the accumulation of potentially harmful protein aggregates. E. faecalis must evolve strategies to combat these stresses encountered during colonization and infection. It is not known how these misfolded proteins, including Ebp, are processed. In this study, we determined the contribution of a highly conserved serine protease, HtrA, to membrane stress tolerance in E. faecalis. We found that HtrAEF is not involved in growth or survival under a wide range of stress conditions, nor is it involved in biofilm formation. However, the ∆htrA mutant was attenuated in a competitive murine wound infection, suggesting its contribution to colonization and infection in host tissues. In many bacteria, HtrA deals with accumulated protein stresses and/or is part of a proteolytic pathway. The absence of HtrA alone does not result in accumulation of proteins. We postulated that the induction of protein accumulation in the absence of HtrA in E. faecalis may reveal a true role for HtrAEF. We created a ∆srtA∆htrA to mimic membrane stress where sortase substrates, such as Ebp pili, accumulate on the cell membrane. In this mutant background, we observed hyperpiliation as compared to single mutants. Furthermore, ∆srtA∆htrA formed chains and displayed aberrant placement of septa in addition to increased pilus expression. ∆srtA∆htrA cells appeared coccoid and wider at the cell width than WT. In addition, this phenotype was pilus-dependent, as we no longer observed these aberrant cell morphologies in a ∆srtA∆ebpABC∆htrA strain. Transcriptomic analysis revealed upregulation of the CroR-CroS two-component system (TCS) in altering cell shape in a ∆srtA∆htrA mutant strain. Deletion of croR in ∆srtA∆htrA resulted in a reversal of cell morphology and piliation level to WT, suggesting that CroR mediates reparative transcriptional responses during membrane stress. Global expression analysis using RNA-seq identified possible CroR target genes, suggesting a possible novel role of this TCS in E. faecalis pilus biogenesis. This thesis is the first comprehensive study of HtrA in E. faecalis. We have discovered novel roles for HtrA and the CroR-CroS TCS in monitoring pilus biogenesis, providing potential inhibitory targets to limit the biogenesis of this important virulence factor. Doctor of Philosophy

Apoptosis of osteoblasts triggered by high-dose glucocorticoids (GCs) has been identified as a major cause of osteoporosis. However, the molecular mechanisms underlying GC-induced osteoporosis remain elusive. This study was conducted to make clear the mechanism of GC-induced osteoblast apoptosis and to examine whether reduction of ER stress by 4-PBA inhibited osteoblast apoptosis. After treatment with dexamethasone (Dex) or hydrocortisone, cell viability was assessed using an MTT assay. Flow cytometry was performed to assess the apoptosis of MC3T3-E1 cells. The expression levels of ER stress-related proteins (CHOP, GRP78, eIF2α, and phospho-eIF2α) and apoptosis-related proteins (cleaved Caspase-3, Bcl-2, and Bax) in MC3T3-E1 cells were measured by Western blot analysis. We found that both Dex and hydrocortisone reduced cell proliferation and promoted apoptosis in MC3T3-E1 cells. In addition, the protein expression levels of cleaved Caspase-3 and Bax increased and the protein expression level of Bcl-2 decreased in MC3T3-E1 cells exposed to Dex. In addition, the Dex exposure also resulted in a release of cytochrome c (Cyt C) from mitochondria. The cellular ATP content was decreased following prolonged treatment with Dex. 4-PBA attenuated ER stress and mitochondrial dysfunction induced by Dex in MC3T3-E1 cells. Dex-mediated apoptosis of MC3T3-E1 cells is aggravated by ER stress. Moreover, Dex-induced apoptosis in MC3T3-E1 cells was inhibited by 4-PBA, suggesting that ER stress involved in Dex-induced apoptosis. In conclusion, inhibition of ER stress by 4-PBA could reduce GC-induced apoptosis in MC3T3-E1 cells.

Receptor activator of nuclear factor-κB ligand (RANKL) is a key factor in the differentiation and activation of osteoclasts. Echinocystic acid (EA), a pentacyclic triterpene isolated from the fruits of Gleditsia sinensis Lam, was reported to prevent reduction of bone mass and strength and improve the cancellous bone structure and biochemical properties in ovariectomy rats. However, the molecular mechanism of EA on the osteoclast formation has not been reported. The purpose of this study was to investigate the effects and mechanism of EA on RANKL-induced osteoclastogenesis. Our results showed that EA inhibited the formation of osteoclast, as well as the expression of osteoclastogenesis-related marker proteins in bone marrow macrophages (BMMs). At molecular levels, EA inhibited RANKL-induced NF-κB activation and ERK phosphorylation in BMMs. In conclusion, the present study demonstrated that EA can suppress osteoclastogenesis in vitro. Moreover, we clarified that these inhibitory effects of EA occur through suppression of NF-κB and ERK activation. Therefore, EA may be a potential agent in the treatment of osteoclast-related diseases such as osteoporosis.

NagR, belonging to the GntR/HutC family, is a negative regulator that directly represses the nagP and nagAB genes, which are involved in GlcNAc transport and utilization in Bacillus subtilis. Our previous work confirmed that the chitinase B gene (chiB) of Bacillus thuringiensis strain Bti75 is also negatively controlled by YvoABt, the ortholog of NagR from B. subtilis. In this work, we investigated its regulatory network in Bti75 and found that YvoABt is an N-acetylglucosamine utilization regulator primarily involved in GlcNAc catabolism; therefore YvoABt is renamed as NagRBt. The RNA-seq data revealed that 27 genes were upregulated and 14 genes were downregulated in the ΔnagR mutant compared with the wild-type strain. The regulon (exponential phase) was characterized by RNA-seq, bioinformatics software, electrophoretic mobility shift assays, and quantitative real-time reverse transcription PCR. In the Bti75 genome, 19 genes that were directly regulated and 30 genes that were indirectly regulated by NagRBt were identified. We compiled in silico, in vitro, and in vivo evidence that NagRBt behaves as a repressor and activator to directly or indirectly influence major biological processes involved in amino sugar metabolism, nucleotide metabolism, fatty acid metabolism, phosphotransferase system, and the Embden–Meyerhof–Parnas pathway.

Orientin is a flavonoid extracted from Chinese traditional herb, Polygonum orientale L. Previous study has reported that orientin protected myocardial from ischemia reperfusion injury. However, whether orientin could protect against cardiac remodeling after myocardial injury remains unclear. The aim of our study is to investigate the effects of orientin in the progression of cardiac remodeling after myocardial infarction (MI). Mice cardiac remodeling model was established by left coronary artery ligation surgery. Experimental groups were as follows: vehicle-sham, orientin-sham, vehicle-MI, and orientin-MI. Animals were treated with vehicle or orientin (40 mg/kg) for 25 days starting 3 days after surgery. After 4 weeks of MI, mice with orientin treatment had decreased mortality and improved cardiac function. Significantly, at 4 weeks post-MI, orientin treatment decreased fibrosis, inflammatory response, and cardiomyocyte apoptosis. Furthermore, orientin treatment attenuated the hypoxia-induced neonatal rat cardiomyocyte apoptosis and increased cell viability. Additionally, orientin supplementation mitigated oxidative stress in remodeling heart tissue and cardiomyocytes exposed to hypoxia as measured by 2′,7′-dichlorodihydrofluorescein diacetate fluorescent probe. Mechanistically, orientin promotes cardioprotection by activating the eNOS/NO signaling cascades, which was confirmed by eNOS inhibitor (L-NAME) in vitro and in vivo. Inhibition of oxidative stress by orientin via eNOS/NO signaling cascades in the heart may represent a potential therapy for cardiac remodeling.

Neuropathic pain (NP) is one of the most common complications after spinal cord injury (SCI), but no protein biomarkers has ever been introduced into clinical diagnosis. Previous studies implicated that toll-like receptor (TLR) 4 played a critical role in the development of NP in animal SCI models. Here, a total of 140 participants were recruited, 70 of them were SCI-NP subject and the rest 70 controls did not show neuropathic symptoms. TLR4 was upregulated significantly in SCI-NP patients compared with SCI-noNP subjects. Furthermore, we measured the concentrations of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6), two TLR4 downstream pro-inflammatory cytokines, to assess their diagnostic values. Receiver operating characteristics (ROC) analysis revealed that TNF-α had great potential advantages to predict the progression of neuropathy, the risks of NP were strongly increased in SCI subjects with higher levels of TNF-α (odds ratio: 4.92; 95% confidence interval: 1.89-12.32). These results suggested neuro-immune activation contributed to the development of neuropathic disorder after SCI, and TNF-α could be a potential sensitive diagnostic biomarker for chronic neuropathic pain in SCI patients.

Acupuncture has been shown to lessen the inflammatory reaction after acute spinal cord injury and reduce secondary injury. However, the mechanism of action remains unclear. In this study, a rat model of spinal cord injury was established by compressing the T 8-9 segments using a modified Nystrom method. Twenty-four hours after injury, Zusanli (ST36), Xuanzhong (GB39), Futu (ST32) and Sanyinjiao (SP6) were stimulated with electroacupuncture. Rats with spinal cord injury alone were used as controls. At 2, 4 and 6 weeks after injury, acetylcholinesterase (AChE) activity at the site of injury, the number of medium and large neurons in the spinal cord anterior horn, glial cell line-derived neurotrophic factor (GDNF) mRNA expression, and Basso, Beattie and Bresnahan locomotor rating scale scores were greater in the electroacupuncture group compared with the control group. These results demonstrate that electroacupuncture increases AChE activity, up-regulates GDNF mRNA expression, and promotes the recovery of motor neuron function in the anterior horn after spinal cord injury.

Background Skin pigmentation is accomplished by production of melanin in melanosome and by transfer of these organelles from melanocytes (MCs) to surrounding keratinocytes (KCs). However, the detailed mechanism is still unknown. Objective We aimed to investigate the morphological structure changes on human epidermal MCs and KCs, which were either mono-cultured or co-cultured, with or without the treatment of both α-Melanocyte-stimulating hormone (α-MSH) and prostaglandin E2 (PGE2), by atomic force microscopy (AFM) and to provide more direct proofs for process of melanosome transfer. Methods Human epidermal MCs and KCs were isolated and co-cultured with 1:10 ratio in a defined Keratinocyte-serum free medium (K-SFM). After exposure with 100nM α-MSH or 20μM PGE2 for 3 days, cells were fixed with 0.5% glutaraldehyde and AFM images of scanning observation were captured by contacting and tapping model under normal atmospheric pressure and temperature. Results It showed that human epidermal MCs in culture had secondary or tertiary branches. Except for globular granules structure on the surface of dendrites, some filopodia were protruded on the tips and lateral sides of the dendrites. The administration of α-MSH and PGE2 made not only the dendrites thinner and longer, but also the globular granules more intensive and denser. Many spheroid granules were shed from branches of dendrite and most of them adhered with dense filopodia. Compared with untreated group, the number of filopodia per cell, diameter of filopodia, and shedding spheroid granules per field all increased following α-MSH and PGE2 exposure ( P n =3). However, many crest-like protrusions, which were distributed homogenously on the surface of mono-cultured KCs, were less changed after α-MSH and PGE2 exposure. In co-culture model, α-MSH and PGE2 increased the number of transferred melanosomes in KCs under laser confocal microscopic examination. Filopodia were observed only on the adhesion area of KCs and MCs in a coiled style by AFM examination. In addition, the number of filopodia per field, diameter of filopodia and shedding spheroid granules per field all increased after the administration of α-MSH and PGE2 ( P n =3). Conclusions Our data suggest that shedding spheroid granules, filopodia delivery and KC phagocytosis are major mode of melanosome transfer between MCs and KCs. PGE2, as well as α-MSH, drives melanosome transfer by promoting filopodia delivery and numbers of shedding spheroid granules in MCs, but no direct morphological effects on KCs. These findings open a new path in our understanding of MCs–KCs communication regulating pigmentation.

This study investigated effects of dietary fibre and grit on growth performance, gastrointestinal tract (GIT) development, serum indexes and grit retention of Sichuan white goslings in China. The experiment was a 3×2 factorial design consisting of dietary crude fibre (CF) at 2.0, 4.0 and 6.0% levels by adding grass meal and with or without grit supplementation. A total of 7201-d-old male goslings were randomly allocated into 36 pens of 20 goslings/pen (n=6 pens/treatment). At 21 d of age, GIT, blood samples and grit in gizzards were collected. Body weight (BW), empty BW (without GIT), average daily gain (ADG) and average daily feed intake (ADFI) were affected by CF levels and the medium (4.0%) had the highest value. Adding grit improved BW, empty BW, ADG and ADFI. CF affected relative weight or length of proventriculus, gizzard and duodenum. Increasing CF levels decreased jejunum villus height and affected caecal crypt depth and villus height to crypt depth ratio. Higher CF levels decreased serum triglyceride. Adding grit increased relative weight of abdominal fat. Grit addition increased grit weight in gizzard and the ratio of grit in 0.45-1 and 1-2 mm while decreasing the ratio of grit

Enterococcus faecalis wound infections remain poorly understood. We demonstrate in vivo that E faecalis infection results in 2 different states: colonization or infection. We show that E faecalis infection modulates host immunity, resulting in persistent infection and delayed healing., Enterococcus faecalis is one of the most frequently isolated bacterial species in wounds yet little is known about its pathogenic mechanisms in this setting. Here, we used a mouse wound excisional model to characterize the infection dynamics of E faecalis and show that infected wounds result in 2 different states depending on the initial inoculum. Low-dose inocula were associated with short-term, low-titer colonization whereas high-dose inocula were associated with acute bacterial replication and long-term persistence. High-dose infection and persistence were also associated with immune cell infiltration, despite suppression of some inflammatory cytokines and delayed wound healing. During high-dose infection, the multiple peptide resistance factor, which is involved in resisting immune clearance, contributes to E faecalis fitness. These results comprehensively describe a mouse model for investigating E faecalis wound infection determinants, and suggest that both immune modulation and resistance contribute to persistent, nonhealing wounds.

The present study investigated the chemical constituents of aerial part of Polygonum pubescens Blume. Twenty-two compounds 1–22 were obtained from petroleum ether and ethyl acetate extracts of aerial part of P. pubescens, including a new phenylpropanoide esters 1 and 21 known compounds. The structures were determined on the basis of spectroscopic and chemical methods. Sixteen compounds were assessed for their cytotoxic and anti-inflammatory activities. Several compounds showed effects on different targets.

It has been proposed that genetic factors contribute to the susceptibility of non-small cell lung cancer (NSCLC). The programmed cell death 6 interacting protein (PDCD6IP) encodes for a protein that has been known to bind to the products of the PDCD6 gene, a required protein in apoptosis. The aim of this study is to investigate the relationship between PDCD6IP insertion/deletion (I/D) polymorphism (rs28381975) and NSCLC risk in a Chinese population. A population-based case–control study was conducted in 449 NSCLC patients and 512 cancer-free controls. The genotype of the PDCD6IP gene was determined by using a polymerase chain reaction assay. The promoter activity was analyzed by luciferase reporter assay in A549 and H1299 cells. Statistically significant difference was observed when the patients and controls were compared according to ID + II versus DD (OR = 1.72, 95 % CI 1.29–2.31, P

The specific recognization between galactose group and Ricinus Communis Agglutinin (RCA) was investigated by microcantilever. The gold side of the microcantilever was covalently bound with N-galactose, RCA and asialofetuin (ASF) via mixed self assembly monolayer of 11-mercaptoundecanoic acid and 6-mercaptohexanol, respectively. After adding RCA into the flowing cell, the deflection could be observed on the N-galactose or ASF modified microcantilever. Meanwhile, the deflection could also be observed after ASF bound to the RCA modified microcantilever. In order to prove that the deflection is caused by the specific interaction between the galactose group and RCA, bovine serum albumin (BSA) was introduced into the flowing cell as control experiment and no obvious deflection was observed. The specific interaction was also confirmed by the evidence that the bound protein layer can be mechanically removed with atomic force microscopy nanolithography technology.

In this study, we investigated the transcriptional response to 50 Hz extremely low frequency electromagnetic field (ELF-EMF) and 2.0 GHz radio frequency electromagnetic field (RF-EMF) exposure by Illumina sequencing technology using budding yeast as the model organism. The transcription levels of 28 genes were upregulated and those of four genes were downregulated under ELF-EMF exposure, while the transcription levels of 29 genes were upregulated and those of 24 genes were downregulated under RF-EMF exposure. After validation by reverse transcription quantitative polymerase chain reaction (RT-qPCR), a concordant direction of change both in differential gene expression (DGE) and RT-qPCR was demonstrated for nine genes under ELF-EMF exposure and for ten genes under RF-EMF exposure. The RT-qPCR results revealed that ELF-EMF and RF-EMF exposure can upregulate the expression of genes involved in glucose transportation and the tricarboxylic acid (TCA) cycle, but not the glycolysis pathway. Energy metabolism is closely related with the cell response to environmental stress including EMF exposure. Our findings may throw light on the mechanism underlying the biological effects of EMF.

The environmental estrogen can be measured in modified yeast cells by transcription activation of reporter genes. To establish such yeast estrogen system, the full-length human estrogen receptor (hER) cDNA and estrogen responsive elements (EREs) linked to the upstream of CYC1 promoter were cloned into the yeast expression vector via pPIC9 vector respectively to construct the response vectors named as pER-PIC9 (E-9) and pERE2-CYC-PIC9K-EGFP (EC-9-E), which were transformed into Pichia pastoris at the same time for detecting environmental estrogens based on yeast Enhanced Green Fluorescence Protein (yEGFP) as a reporter protein. In the present study, we developed a rapid yeast estrogen bioassays by creating stably transfected strains with EGFP as a measurable reporter protein.

Introgression lines are important genetic materials for genetics study and breeding. Development of those lines involves cross and backcross processes between recipient and donor parents. The population size of BC1F1 is a critical parameter for fully covering donor genome and successfully obtaining desired introgression lines. However, the minimum sufficient number of BC1F1 plants is unknown for each species and cannot be obtained experimentally. A computer program was developed by simulating the recombination process during meiosis to define the ideal BC1F1 population size. The reliability of the program was confirmed by mathematics and experimental data. Three factors including linkage group number, linkage group length, and gene density were analyzed, and all of them had positive relation with the size of BC1F1 population. The population size increased from 6.06 to 9.49 when the linkage number increased from 5 to 40. The population size was 7.14 when the linkage group length was 80 cM, while it became 8.64 when the length was 200 cM. The population size was 7.65 with the density of 20 cM per gene and 8.22 with 10 cM per gene. The BC1F1 population sizes of rice (Oryza sativa L.), wheat (Triticum aestivum L.), maize (Zea mays L.), and soybean (Glycine max L. Merr.) were predicted to be 12, 13, 14–15, and 13, respectively, by the program with 95% confidential level.

Mantle cell lymphoma (MCL) is a rare, specific lymphoma subtype. Though the morphologic and immunophenotypic features of MCL have been well described in recent literatures, it is still a diagnostic dilemma because of its frequent confusion with other small B cell lymphomas (SBCLs). In the present study, we primarily focus on establishing a sensitive and specific method for the diagnosis of MCL, which is efficient to distinguish this disease from other SBCLs. We carried out our investigation for MCL and other SBCLs (including SLL, FL, MZL, and MALT) on their feature of morphology, immunophenotype, and t(11;14)(q13;q32) translocation analysis based on polymerase chain reaction (PCR) and interphase nuclei micro-array fluorescence in situ hybridization (FISH). The morphologic and immunologic analysis showed the positive rate of cyclin D1 was 76.47% in MCL, which was significantly higher than that in other SBCLs. The positive rate of t(11;14) translocation was 25.81% and 35.48%, respectively, tested by general and semi-nested PCR, while 93.10% positive rate was shown with low background and strong signals pattern when tested by Nuclei micro-array FISH. Our research shows that t(11;14) translocation is a special and useful diagnostic marker for MCL, and detection of the marker by nuclei micro-array FISH is convenient and economic, especially more sensitive and specific than other methods for the diagnosis of MCL.

In an earlier study, an expressed sequence tag (EST) from a drought-related suppression subtractive hybridization (SSH) library was obtained, which had a high similarity to AtCKS2 from Arabidopsis . To dissect the structure and expression characteristic of this EST, the cDNA homologous to CKS2 was isolated using in silico and homology-based cloning techniques, and which was designated ZmCKS2 . ZmCKS2 has an open reading frame of 267 bp and encodes 88 amino acids. The deduced protein of ZmCKS2 is predicted to contain a cyclin-dependent kinase regulatory subunit (CKS) structural domain, which was highly conserved in eukaryotes kingdom. The promoter of ZmCKS2 (about 2 kb upstream of ZmCKS2 ) was predicted to contain important regulatory elements including core promoter elements and drought, high salt, coldness, abscisic acid (ABA), light regulation, copper, and oxygen-responsive elements. Real-time quantitative PCR (qRT-PCR) result revealed that ZmCKS2 had different expression patterns in different organs. It was highly expressed in ears, while its expression was very low in roots and leaves at seedling stage. qRT-PCR was also performed to investigate the expression profiles of ZmCKS2 under different abiotic stresses such as drought, low temperature, high salt, methyl jasmonate (MeJA), and exogenous ABA. Under drought or MeJA treatment, ZmCKS2 showed an up-regulated expression pattern. However, under cold or ABA treatment, this gene showed a down-regulation expression pattern. Under NaCl stress ZmCKS2 showed no obvious change. These results revealed that ZmCKS2 might be involved in multiple pathways of plants responding to different environmental conditions.

Denaturation and aggregation of proteins are reactions that are relevant to functional applications of proteins in foods. Depending on concentration, ionic strength, and pH, aggregation can result in turbidity, precipitation, or gelation. Aggregation may be desirable, as in the case of gelation, or undesirable, as in the case when it causes phase separation in beverages. One approach to improve the stability of globular proteins against heat stresses is through the addition of other compounds that alter aggregation. Numerous studies have shown the ability of molecular chaperones to assist proper folding/unfolding and assembly/disassembly of proteins, especially during stressed conditions. Recently, several papers have reported the molecular chaperone-like properties of caseins, especially using alpha(s)- and beta-caseins. Caseins appear to function like small heat shock proteins (sHSP). We have compared the results among investigations from the perspective of food processing conditions and related them to the mechanism for sHSP. Caseins possess three of the four common features among sHSP; lacking a similar sequence domain. Their function may be explained in part by having structures fitting the intrinsically unfolded class of proteins. With a few exceptions, most investigations were done at solution conditions that poorly represent foods; lacking investigations at pH4.5 and concentrations above 20 mg/mL. While it is clear that caseins can alter aggregation at neutral pH, their effectiveness at low pH, high protein concentration, and high thermal treatment (Tor= 100 degrees C) remains to be fully established.

Ubiquitin linkage to cysteine is an unconventional modification targeting protein for degradation. However, the physiological regulation of cysteine ubiquitination is still mysterious. Here we found that ACAT2, a cellular enzyme converting cholesterol and fatty acid to cholesteryl esters, was ubiquitinated on cysteine 277 (C277) for degradation when lipid level was low. gp78/Insigs catalyzed K48-linked polyubiquitination on this C277. High concentration of cholesterol and fatty acid, however, induced cellular reactive oxygen species (ROS) that oxidized C277, rendering ACAT2 stabilization and subsequently elevated cholesteryl esters. Furthermore, ACAT2 knockout mice were more susceptible to HFD-associated insulin resistance. By contrast, expression of constitutively stable form of ACAT2 (C277A) resulted in higher insulin sensitivity. Together, these data indicate that lipid-induced stabilization of ACAT2 ameliorates lipotoxicity from excessive cholesterol and fatty acid. This unconventional cysteine ubiquitination of ACAT2 constitutes an important mechanism sensing lipid overload-induced ROS and fine-tuning lipid homeostasis.

Maize roots are important component for plant adaptation to soil water deficits because they are supposed to take up water and necessary solutes from the soil. In the present study, the drought-induced genes were isolated in maize roots. A suppression subtractive hybridization protocol was applied to construct a forward subtractive cDNA library from CN165 for drought-stressed maize roots and a number of drought-induced genes were isolated. Totally, 126 uniESTs (containing 82 singlets and 44 contigs) were obtained from 503 available ESTs sequences after macroarray hybridization. UniESTs were analyzed using BLASTN and BLASTX and the results showed that 92% of the uniESTs had homolgous sequences in maize nr database by BLASTN. About 89% of uniESTs appeared the homlogous amino acid sequences in rice protein database but not in maize protein database by BLASTX, implying that those genes are likely new functional genes in maize. Function analysis showed that those genes were involved in a broad spectrum of biological pathways, mainly in signaling and regulatory pathways related to stress tolerance.

Sex determination mechanisms are thought to evolve rapidly and show little conservation among different animal species. For example, the critical gene on the Y chromosome, SRY, that determines sex in most mammals, is not found in other animals. However, a related Sox domain transcription factor, SOX9, is also required for testis development in mammals and exhibits male-specific gonad expression in other vertebrate species. Previously, we found that the Drosophila orthologue of SOX9, Sox100B, is expressed male-specifically during gonad development. We now investigate the function of Sox100B and find, strikingly, that Sox100B is essential for testis development in Drosophila. In Sox100B mutants, the adult testis is severely reduced and fails to interact with other parts of the reproductive tract, which are themselves unaffected. While a testis initially forms in Sox100B mutants, it fails to undergo proper morphogenesis during pupal stages, likely due to defects in the pigment cells. In contrast, no substantive defects are observed in ovary development in Sox100B mutant females. Thus, as is observed in mammals, a Sox9 homolog is essential for sex-specific gonad development in Drosophila, suggesting that the molecular mechanisms regulating sexually dimorphic gonad development may be more conserved than previously suspected.

To localize the pathogenic genes of autosomal dominant ichthyosis vulgaris, we ascertained two ichthyosis vulgaris families from Hunan Province. Venous blood samples were collected from affected and unaffected family members and genomic DNA was extracted. We then performed genome scan and linkage analysis using microsatellite markers around known ichthyosis vulgaris loci in chromosomes 1 and 10. In family 1, the locus linked to ichthyosis vulgaris was located near D1S498 (1q21), which overlapped with known ichthyosis vulgaris loci. In family 2, however, all known loci for ichthyosis vulgaris were excluded and the new locus remains to be identified.

In this study, a forward cDNA library was constructed by suppression subtractive hybridization using seedling leaves of CN165, a drought-tolerant maize inbred line. In the suppression subtractive hybridization (SSH) library, 672 positive clones were picked up randomly. After polymerase chain reaction (PCR) of each clone, all the single clones were sequenced. Totally 598 available sequences were obtained. After cluster analysis of the EST sequences, 80 uniESTs were obtained, among which 57 uniESTs were contigs and 23 uniESTs were singlets. The results of BLASTN showed that all the uniESTs had homologous sequences in the nr database. The BLASTX results indicated that 68 uniESTs had significant protein homology, 8 uniESTs with homology of unknown proteins and putative proteins, and 4 uniESTs without protein homology. Those drought stress-induced genes were involved in many metabolism pathways to regulate plant growth and development under drought stress.

Maize female organs are sensitive to drought stress, leading to reproductive failure and yield reduction. In the present study gene expression profiles of ears and silks of maize at the flowering stage under drought stress were investigated. From 1920 white positive clones of a forward suppression subtractive hybridization (SSH) library, 1439 available sequences of expression sequence tags (ESTs) were obtained, resulting in 361 unique ESTs after assembling. Data analysis showed that 218 of the unique ESTs had significant protein homology by BLASTX in UNIPROT database. Totally 99 uniESTs were found in TIGR maize gene indices and nr database by BLASTN, while 44 uniESTs were not found to have homologous nucleic acid sequences and putatively classified as "maize-specific" uniESTs. The 218 cDNAs with significant protein homology were sorted into 13 groups according to the functional categories of the Arabidopsis proteins. Among those genes, the genes associated with the metabolisms were the largest group (account for 27%), and the genes related to protein synthesis, protein fate, transcription, cell cycle and DNA processing accounted for 16, 10, 10 and 9%, respectively. After analysis of macroarray data and real-time quantitative polymerase chain reaction (PCR), it was found that 160 of the 218 homologous protein uniESTs were up-regulated genes in the ears, 129 in the silks, and 125 in both of the tissues. The present work provided a valuable starting point for further elucidation of the roles played by these genes/gene products in drought tolerance in maize.

We construct the bounce-averaged diffusion coefficients and study the bounce-averaged acceleration for energetic electrons in gyroresonance with whistler mode chorus. Numerical calculations have been performed for a band of chorus frequency distributed over a standard Gaussian spectrum specifically in the region near L = 4.5, where peaks of the electron phase space density occur. It is found that whistler mode chorus can efficiently accelerate electrons and can increase the phase space density at energies of about 1 MeV by more than one order of magnitude about one day, in agreement with the satellite observations during the recovery phase of magnetic storms.

IMM-H004, a 3-piperazinylcoumarin compound derived from coumarin, has been proved effective against CA1 cell loss and spatial learning impairments resulting from transient global ischemia/reperfusion (TGCI/R), while the mechanism is still largely unknown. Here, we confirmed that treatment of rats with IMM-H004 immediately after TGCI/R ameliorated delayed neuronal death (DND) in the CA1 of hippocampus and cortex. Further study suggested that IMM-H004 contributed to the expression of antiapoptotic protein survivin through the activation of PI3K-dependent protein kinase B (PKB/Akt), which led to the phosphorylation of forkhead box O1 (FoxO1), then relieved the inhibiting effect on survivin promoter. Additionally, IMM-H004 also enhanced the expression of hepatitis B X-interacting protein (HBXIP), which formed a complex with survivin to prevent the activation of caspase death cascade, thereby halting apoptotic cell death. Finally, we injected a HBXIP siRNA into hippocampus and performed microelectroporation before ischemia/reperfusion, which abolished the protective effect of IMM-H004. Further study revealed that HBXIP maintained the high expression of Akt and survivin. Collectively, our findings demonstrated that DND after TGCI/R was alleviated by IMM-H004 through promoting the formation of survivin-HBXIP complex, which further emphasized the importance of endogenous protein involved in self-repair after stroke.

Background: The insulin-like growth factor (IGF) system comprises a group of proteins that play key roles in regulating cell growth, differentiation, and apoptosis in a variety of cellular systems. The aim of this study was to investigate the role of insulin-like growth factor binding protein 3 (IGFBP3) in hepatocellular carcinoma. Materials and Methods: Expression of IGF2, IGFBP3, and PTEN was analyzed by qRT-PCR. Lentivirus vectors were used to overexpress IGFBP3 in hepatocellular carcinoma cell (HCC) lines. The effect of IGFBP3 on proliferation was investigated by MTT and colony formation assays. Results: Expression of IGF2, IGFBP3, and PTEN in several HCC cell lines was lower than in normal cell lines. After 5-aza-2’-deoxycytidine/trichostatin A treatment, significant demethylation of the promoter region of IGFBP3 was observed in HCC cells. Overexpression of IGFBP3 induced apoptosis and reduced colony formation in HUH7 cells. Conclusions: Expression of IGF2, IGFBP3, and PTEN in several HCC cell lines was lower than in normal cell lines. After 5-aza-2’-deoxycytidine/ trichostatin A treatment, significant demethylation of the promoter region of IGFBP3 was observed in HCC cells. Overexpression of IGFBP3 induced apoptosis and reduced colony formation in HUH7 cells.

The mammalian liver has a very strong regeneration capacity after partial hepatectomy (PH). To further learn the genes participating in the liver regeneration (LR), 551 cDNAs selected from subtracted cDNA libraries of the regenerating rat liver were screened by microarray, and their expression profiles were studied by cluster and generalization analyses. Among them, 177 genes were identified unreported and up- or down-regulated more than twofold at one or more time points after PH, of which 62 genes were down-regulated to less than 0.5; 99 genes were up-regulated to 2–10 folds, and 16 genes were either up- or down-regulated at different time points during LR. By using BLAST and GENSCAN, these genes were located on responsible chromosomes with 131 genes on the long arms of the chromosomes. The cluster and generalization analyses showed that the gene expression profiles are similar in 2 and 4, 12 and 16, 96 and 144 h respectively after PH, suggesting that the actions of the genes expressed in the same profiles are similar, and those expressed in different profiles have less similarity. However, the types, characteristics and functions of the 177 genes remain to be further studied.

The endothelial cell line ECV304, derived from human umbilical cord and identified to be susceptible to dengue virus type 2 (DEN-2) infection, was used to study the molecular mechanism of DEN-2 binding to endothelial cells. DEN-2 was found by virus overlay protein-binding assays (VOPBAs) to bind to three ECV304 cell membrane proteins with molecular masses of 29, 34 and 43 kDa. Only a single protein of 29 kDa was observed when VOPBAs were carried out using preparations of trypsin-treated ECV304 cells. Pre-incubation of live ECV304 cells in culture or cell membrane proteins in modified VOPBAs with the recombinant DEN-2 envelope glycoprotein (rEgp) inhibited DEN-2 infection and blocked virus binding to the three proteins identified. These results indicate that DEN-2 rEgp could bind to three proteins on the surface of ECV304 cells. This virus-cell interaction may be associated with the receptor complex specific for DEN-2 infection of endothelial cells.