ABSTRACT
Lilium pumilum DC (L. pumilum DC) plays an important role in the rational utilization of salinized soil. To explore the molecular mechanism of salt-tolerant L. pumilum, the LpMYB4 was cloned. LpMYB4 close relationship with Bambusa emeiensis and Zea mays MYB4 throughout the phylogenetic tree construction. LpMYB4 protein was found to be localized in the nucleus. Prokaryotic and eukaryotic bacterial solution resistance experiments proved that the exogenous introduction of LpMYB4 made the overexpression strains obtain better survival ability under saline-alkaline stress. Compared with wild-type plants, tobacco plants overexpressing LpMYB4 had better growth and lower leaf wilting and lodging, the content of chlorophyll was higher, the content of hydrogen peroxide and superoxide anion was lower, the activity of peroxidase and superoxide dismutase was higher and the relative conductivity was lower under saline-alkaline stress. The analysis of seed germination and seedling resistance of transgenic plants under salt stress showed that LpMYB4 transgenic seeds were more tolerant to salt stress during germination and growth. Yeast two-hybrid and two-luciferase complementation experiments showed that LpMYB4 interacted with yeast two-hybrid and LpGPX6. The analysis of the role of LpMYB4 in improving plant saline-alkali resistance is helpful to the transformation of plant germplasm resources and has great significance for agriculture and sustainable development.
Subject(s)
Lilium , Plant Proteins , Plants, Genetically Modified , Salt Tolerance , Plant Proteins/metabolism , Plant Proteins/genetics , Plants, Genetically Modified/genetics , Lilium/genetics , Lilium/metabolism , Salt Tolerance/genetics , Gene Expression Regulation, Plant , Phylogeny , Alkalies , Nicotiana/genetics , Nicotiana/metabolism , Transcription Factors/metabolism , Transcription Factors/genetics , Germination/genetics , Stress, Physiological/geneticsABSTRACT
BACKGROUND: Head and neck squamous cell carcinoma (HNSCC) is the sixth most common malignancy worldwide, characterized by high morbidity, high mortality, and poor prognosis. Collagen triple helix repeat containing 1 (CTHRC1) has been shown to be highly expressed in various cancers. However, its biological functions, potential role as a biomarker, and its relationship with immune infiltrates in HNSCC remain unclear. Our principal objective was to analyze CTHRC1 expression, its prognostic implications, biological functions, and its effects on the immune system in HNSCC patients using bioinformatics analysis. METHODS: The expression matrix was obtained from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO). CTHRC1 expression in HNSCC was analyzed between tumor and adjacent normal tissues, different stages were compared, and its impact on clinical prognosis was assessed using Kaplan-Meier analysis. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Set Variation Analysis (GSVA) were employed for enrichment analysis. The Search Tool for the Retrieval of Interacting Genes database (STRING) was used to analyze protein-protein interactions. Pearson correlation tests were used to investigate the association between CTHRC1 expression and immune checkpoints. The correlation between CTHRC1 and immune infiltration was investigated using CIBERSORT, TIMER, and ESTIMATE. RESULTS: Compared to adjacent normal tissues, CTHRC1 was found to be highly overexpressed in tumors. Increased expression of CTHRC1 was more evident in the advanced stage of HNSCC and predicted a poor prognosis. Most genes related to CTHRC1 in HNSCC were enriched in physiological functions of Extracellular matrix(ECM) and tumor. Furthermore, several immune checkpoints, such as TNFSF4 and CD276 have been shown to be associated with CTHRC1 expression. Notably, the level of CTHRC1 expression correlated significantly with immune infiltration levels, particularly activated macrophages in HNSCC. CONCLUSIONS: High expression of CTHRC1 predicts poor prognosis and is associated with immune infiltration in HNSCC, confirming its utility as a tumor marker for HNSCC. TRIAL REGISTRATION: Not applicable. All data are from public databases and do not contain any clinical trials.
Subject(s)
Biomarkers, Tumor , Extracellular Matrix Proteins , Head and Neck Neoplasms , Squamous Cell Carcinoma of Head and Neck , Humans , Biomarkers, Tumor/metabolism , Prognosis , Squamous Cell Carcinoma of Head and Neck/immunology , Squamous Cell Carcinoma of Head and Neck/genetics , Extracellular Matrix Proteins/genetics , Extracellular Matrix Proteins/metabolism , Head and Neck Neoplasms/immunology , Head and Neck Neoplasms/genetics , Head and Neck Neoplasms/metabolism , Computational Biology , MaleABSTRACT
OBJECTIVES: The study was to explore the causal effects of sleep characteristics on temporomandibular disorder (TMD)-related pain using Mendelian randomization (MR) analysis. MATERIALS AND METHODS: Five sleep characteristics (short sleep, insomnia, chronotype, snoring, sleep apnea) were designated as exposure factors. Data were obtained from previous publicized genome-wide association studies and single nucleotide polymorphisms (SNPs) strongly associated with them were utilized as instrumental variables (IVs). TMD-related pain was designed as outcome variable and sourced from the FinnGens database. MR analysis was employed to explore the causal effects of the five sleep characteristics on TMD-related pain. The causal effect was analyzed using the inverse variance-weighted (IVW), weighted median, and MR-Egger methods. Subsequently, sensitivity analyses were conducted using Cochran's Q tests, funnel plots, leave-one-out analyses, and MR-Egger intercept tests. RESULTS: A causal effect of short sleep on TMD-related pain was revealed by IVW (OR: 1.60, 95% CI: 1.06-2.41, P = 0.026). No causal relationship was identified between other sleep characteristics (insomnia, chronotype, snoring, sleep apnea) and TMD-related pain. CONCLUSIONS: Our study suggests that short sleep may increase the risk of TMD-related pain, while there was no causal relationship between other sleep characteristics and TMD-related pain. Further studies are warranted to deepen and definitively clarify their relationship. CLINICAL RELEVANCE: These findings reveal that the short sleep may be a risk factor of TMD-related pain and highlight the potential therapeutical effect of extending sleep time on alleviating TMD-related pain.
Subject(s)
Mendelian Randomization Analysis , Polymorphism, Single Nucleotide , Temporomandibular Joint Disorders , Humans , Temporomandibular Joint Disorders/genetics , Genome-Wide Association Study , Risk Factors , Snoring , Sleep Wake Disorders/genetics , Sleep Apnea Syndromes/geneticsABSTRACT
Ferric reduction oxidase (FRO) plays important roles in biotic and abiotic stress. However, the function of ferric reduction oxidase from Lilium pumilum in response to NaHCO3 is unknown. Here we report the functional characterization of ferric reduction oxidase 7 in Lilium pumilum (LpFRO7) in stresses. Under NaHCO3 stress, the LpFRO7 overexpression lines exhibited lower accumulation of reactive oxygen species (ROS), higher activities in antioxidant enzyme (CAT, SOD and POD) and ferrite reductase, resulting in improved tolerance compared to the wild type (WT). In order to determine the functional network of LpFRO7, it was confirmed by EMSA assays, Yeast one-hybrid assays and Dual luciferase reporter assays that LpbHLH115 transcription factor can bind to the promoter of LpFRO7. Yeast two-hybrid assays, BiFC, and LCI assays were performed to prove that LpFRO7 can interact with LpTrx. Combining these findings, we concluded that LpFRO7 affects plant saline-alkaline tolerance by regulating ROS homeostasis.
Subject(s)
Lilium , Reactive Oxygen Species/metabolism , Lilium/genetics , Oxidoreductases/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Stress, Physiological , Plants, Genetically Modified/metabolism , Homeostasis , Gene Expression Regulation, PlantABSTRACT
(1) Background: The growth of plants is impacted by salinity and alkali, Lilium pumilum (L. pumilum) is an ornamental plant with strong resistance to salinity and alkali, while the LpPsbP gene is helpful to fully understand the Saline-Alkali tolerance of L. pumilum. (2) Methods: Gene cloning, bioinformatics analysis, expression of fusion protein, determination of physiological indices of plant after Saline-Alkali stress, yeast two-hybrid screening, luciferase complementation assay, chromosome walking to obtain the promoter sequence, and then analyzed by PlantCARE. (3) Results: The LpPsbP gene was cloned and the fusion protein was purified. The transgenic plants had higher Saline-Alkali resistance than the wild type. A total of eighteen proteins interacting with LpPsbP were screened, and nine sites in the promoter sequence were analyzed. (4) Conclusion: Under Saline-Alkali or oxidative stress, L. pumilum will promote the expression of LpPsbP, which will then directly scavenge reactive oxygen species (ROS) in order to protect its photosystem II, reduce its damage, and thus improve the Saline-Alkali resistance of the plant. Moreover, according to some of the literature and the following experiments, two additional speculations are developed on the mechanisms of how two newly found objects, namely jasmonic acid (JA) and FoxO protein, could be involved in ROS scavenging processes were made.
Subject(s)
Lilium , Reactive Oxygen Species/metabolism , Lilium/genetics , Alkalies/metabolism , Oxidative Stress , Plants, Genetically Modified/genetics , Plant Proteins/genetics , Stress, Physiological/genetics , Gene Expression Regulation, PlantABSTRACT
The autophagy adaptor protein SQSTM1/p62 is overexpressed in breast cancer and has been identified as a metastasis-related protein. However, the mechanism by which SQSTM1/p62 contributes to breast cancer progression and tumor microenvironment remains unclear. This study revealed that silencing SQSTM1/p62 expression suppressed breast cancer progression via regulating cell proliferation and reshaping the tumor microenvironment (TME). Here, we found that SQSTM1/p62 was overexpressed in multiple human cancer tissue types and that was correlated with poor patient overall survival (OS) and disease-free survival (DFS). Moreover, we found that short-hairpin RNA (shRNA)-mediated knockdown of p62 expression significantly inhibited cell proliferation, migration, and invasion, and promoted cell death in vitro, as well as suppressed breast cancer growth and lung metastasis in vivo. In addition, flow cytometry analysis of splenocytes and tumor infiltrating lymphocytes (TILs) indicated that the numbers of CD8α+ interferon (IFN)-γ+ cells (CTLs) and CD4+IFN-γ+ (Th1) cells were increased while those of CD4+IL-4+ (Th2) cells, tumor-associated macrophages (TAMs) and myeloid-derived suppressor cells (MDSCs) were decreased. RT-PCR analyses showed that the gene expression of Th1/Th2 cytokines changed in the tumor microenvironment. Silencing SQSTM1/p62 suppressed tumor cell lung metastasis. Together, our results provide strong evidence that silencing tumor cell SQSTM1/p62 inhibited tumor growth and metastasis through cell cycle arrest and TME regulation. This finding provides a novel molecular therapeutic strategy for breast cancer progression and metastasis treatment.
ABSTRACT
Lilium Pumilum with wide distribution is highly tolerant to salinity. The blue copper protein LpCPC (Lilium pumilum Cucumber Peeling Cupredoxin) gene was cloned from Lilium pumilum, which has the conserved regions of type I copper protein. Moreover, LpCPC has the closest relation to CPC from Actinidia chinensis using DNAMAN software and MEGA7 software. qRT-PCR indicated that LpCPC expression was higher in root and bulb of Lilium pumilum, and the expression of the LpCPC gene increased and reached the highest level at 12 h in bulbs under 20 mM NaHCO3. The transgenic yeast was more tolerant compared with the control under NaHCO3 stress. Compared with the wild type, overexpressing plants indicated a relatively lower degree of wilting. In addition, the chlorophyll content, soluble phenol content, and lignin content of overexpressing lines were higher than that of wild-type, whereas the relative conductivity of overexpressing plants was significantly lower than that of wild-type plants. Expression of essential genes including NHX1 and SOS1 in salt stress response pathways are steadily higher in overexpression tobacco than that in wild-types. Transgenic lines had much higher levels of CCR1 and CAD, which are involved in lignin production, compared with wild-type lines. The yeast two-hybrid technique was applied to screen probable interacting proteins interacting with LpCPC. Eight proteins interacted with LpCPC were screened, and five of which were demonstrated to be associated with plant salinity resistance. Overall, the role of gene LpCPC is mediating molecule responses in increasing saline-alkali stress resistance, indicating that it is an essential gene to enhance salt tolerance in Lilium pumilum.
Subject(s)
Lilium , Alkalies/metabolism , Copper/metabolism , Gene Expression Regulation, Plant/genetics , Lignin/metabolism , Lilium/genetics , Lilium/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Saccharomyces cerevisiae/metabolismABSTRACT
slPHB3 was cloned from Salix linearistipularis, the amino acid sequence blast and phylogenetic tree analysis showed that slPHB3 has the most similarity with PHB3 from Populus trichocarpa using DNAMAN software and MEGA7 software. RT-qPCR results confirmed that the expression of slPHB3 was induced obviously under stress treatments. The growth of recombinant yeast cells was better than that of the control group under the stress treatment, indicating that slPHB3 may be involved in the stress response of yeast cells. The transgenic tobacco was treated with different concentrations of NaCl, NaHCO3 and H2O2, fresh weigh of overexpression tobacco were heavier than wild-types. The results showed that transgenic tobacco was more tolerant to salt and oxidation than wild-type tobacco. Expression of important genes including NHX1 and SOS1 in salt stress response pathways are steadily higher in overexpression tobacco than that in wild-types. We identified 17 proteins interacting with slPHB3 by yeast two-hybrid technique, most of these proteins were relation to the stresses. The salt tolerance of slPHB3 expressing yeast and slPHB3 overexpressing plants were better than that of the control. Ten stress-related proteins may interact with slPHB3, which preliminarily indicated that slPHB3 had a certain response relationship with salt stress. The study of slPHB3 under abiotic stress can improve our understanding of PHB3 gene function.
Subject(s)
Gene Expression Regulation, Plant , Plant Proteins , Hydrogen Peroxide/metabolism , Phylogeny , Plant Proteins/genetics , Plant Proteins/metabolism , Plants, Genetically Modified/metabolism , Saccharomyces cerevisiae/metabolism , Stress, Physiological/genetics , Nicotiana/metabolismABSTRACT
In the past, the PHB gene function was mainly focused on anti-cell proliferation and antitumor effects. But the molecular mechanism of the PHB gene regarding saline and oxidative stresses is unclear. To study the role of AtPHB6 in salt and oxidative stress, AtPHB6 was cloned from A. thaliana. Bioinformatics analysis showed that AtPHB6 was closely related to AtPHB1 and AtPHB2, which are both type II PHB. RT-qPCR results indicated that the AtPHB6 in the leaves and roots of A. thaliana was obviously induced under different stress treatments. AtPHB6-overexpressing plants were larger and more lush than wild-type and mutant plants when placed under stress treatments during seed germination. The root length and fresh weight of AtPHB6 transgenic plants showed the best resistance compared to wild-type plants under different treatments, in contrast, the AtPHB6 mutants had the worst resistance during the seedling stage. AtSOT12 was an interacting protein of AtPHB6, which screened by yeast two-hybrid system. The interaction between the two proteins were further confirmed using in vitro pull-down experiments and in vivo BiFC experiments. Subcellular localization showed both AtPHB6 and AtSOT12 protein expressed in the nucleus and cytoplasm. The H2O2 content in both the transgenic AtPHB6 and AtSOT12 plants were lower than that in the wild type under stresses. Thus, AtPHB6 increased plant resistance to salt stress and interacted with the AtSOT12 protein.
Subject(s)
Arabidopsis , Arabidopsis/genetics , Arabidopsis/metabolism , Gene Expression Regulation, Plant , Hydrogen Peroxide , Plant Proteins/genetics , Plant Proteins/metabolism , Plants, Genetically Modified/metabolism , Reactive Oxygen Species/metabolism , Stress, PhysiologicalABSTRACT
The formation of neutrophil extracellular traps (NETs) was recently identified as one of the most important processes for the maintenance of host tissue homeostasis in bacterial infection. Meanwhile, pneumonia infection has a poor effect on cancer patients receiving immunotherapy. Whether pneumonia-mediated NETs increase lung metastasis remains unclear. In this study, we identified a critical role for multidrug-resistant Staphylococcus aureus infection-induced NETs in the regulation of cancer cell metastasis. Notably, S. aureus triggered autophagy-dependent NETs formation in vitro and in vivo and increased cancer cell metastasis. Targeting autophagy effectively regulated NETs formation, which contributed to the control of cancer metastasis in vivo. Moreover, the degradation of NETs by DNase I significantly suppresses metastasis in lung. Our work offers novel insight into the mechanisms of metastasis induced by bacterial pneumonia and provides a potential therapeutic strategy for pneumonia-related metastasis.
ABSTRACT
Necroptosis is a form of programmed cell death (PCD) characterized by RIP3 mediated MLKL activation and increased membrane permeability via MLKL oligomerization. Tumor cell immunogenic cell death (ICD) has been considered to be essential for the anti-tumor response, which is associated with DC recruitment, activation, and maturation. In this study, we found that P. aeruginosa showed its potential to suppress tumor growth and enable long-lasting anti-tumor immunity in vivo. What's more, phosphorylation- RIP3 and MLKL activation induced by P. aeruginosa infection resulted in tumor cell necrotic cell death and HMGB1 production, indicating that P. aeruginosa can cause immunogenic cell death. The necrotic cell death can further drive a robust anti-tumor response via promoting tumor cell death, inhibiting tumor cell proliferation, and modulating systemic immune responses and local immune microenvironment in tumor. Moreover, dying tumor cells killed by P. aeruginosa can catalyze DC maturation, which enhanced the antigen-presenting ability of DC cells. These findings demonstrate that P. aeruginosa can induce immunogenic cell death and trigger a robust long-lasting anti-tumor response along with reshaping tumor microenvironment.
ABSTRACT
BACKGROUND: Tachyplesin III, an antimicrobial peptide (AMP), provides protection against multidrug-resistant (MDR) bacterial infections and shows cytotoxicity to mammalian cells. Mixed bacterial infections, of which P. aeruginosa plus A. baumannii is the most common and dangerous combination, are critical contributors to the morbidity and mortality of long-term in-hospital respiratory medicine patients. Therefore, the development of effective therapeutic approaches to mixed bacterial infections is urgently needed. METHODS AND RESULTS: In this study, we demonstrated that compared with individual infections, mixed infections with MDR bacteria P. aeruginosa and A. baumannii cause more serious diseases, with increased pro-inflammatory cytokines (IL-1ß, IL-6, TNF-α) and chemokines (MCP-1/MIP-2) and reduced mouse survival. In vitro treatment with Tachyplesin III enhanced phagocytosis in a mouse alveolar macrophage cell line (MH-S). Strikingly, in vivo, Tachyplesin III demonstrated a potential role against mixed-MDR bacterial coinfection. The bacterial burden in bronchoalveolar lavage fluid (BALF) was significantly reduced in the Tachyplesin III-treated group. In addition, a systemic reduction in pro-inflammatory cytokines and decreased lung injury occurred with Tachyplesin III therapy. CONCLUSION: Therefore, our study demonstrated that Tachyplesin III represents a potential therapeutic treatment against mixed-MDR bacterial infection in vivo, which sheds light on the development of therapeutic strategies against mixed-MDR bacterial infections.
ABSTRACT
Although the antimicrobial peptide cathelicidin-BF shows minimal cytotoxicity in mammalian cells and has excellent direct killing effects on multidrug-resistant clinical pathogens such as Pseudomonas aeruginosa, its clinical application is precluded by its high sensitivity to serum proteases. Here, we demonstrate that intravenous administration of cathelicidin-BF after P. aeruginosa infection did not increase the survival rate of mice with acute pneumonia but that pretreatment with cathelicidin-BF ameliorated pneumonia by effectively activating innate immunity. Enhanced neutrophil extracellular trap (NET) activation and release (NETosis) are key processes for capturing and killing bacteria, concomitantly enhanced macrophage clearance activity, including phagocytosis and autophagy, may eliminate NETs early enough to prevent severe tissue damage. Our study not only suggests a possible approach for applying cathelicidin-BF in vivo but also provides a possible defense strategy against multidrug-resistant pathogens, i.e., efficiently activation of innate immunity.
Subject(s)
Cathelicidins/therapeutic use , Extracellular Traps/physiology , Macrophages/physiology , Neutrophils/physiology , Pneumonia/drug therapy , Pseudomonas Infections/drug therapy , Pseudomonas aeruginosa/physiology , Animals , Autophagy , Disease Models, Animal , Female , Humans , Immunity, Innate , Mice , Mice, Inbred BALB C , Neutrophil Infiltration , PhagocytosisABSTRACT
A type 2 metallothionein gene, SsMT2, was cloned from Suaeda salsa, a salt- and alkali-tolerant plant, which is dominant species on the saline/alkali soil of northeast China. The SsMT2 gene was expressed in all organs except the flower and its expression was induced by various stresses such as CdCl2, NaCl, NaHCO3, and H2O2 treatments. SsMT2-transgenic yeast (Saccharomyces cerevisiae) and plants (Arabidopsis thaliana) showed significantly increased resistance to metal, salt and oxidant stresses. These transgenics accumulated more Cd2+, but less Na+ than their wild type counterparts. SsMT2 transgenic Arabidopsis maintained lower level of H2O2 than wild type plants did in response to the stress treatments. These results demonstrated that the SsMT2 gene plays an important role in reactive oxygen species scavenging and confers enhanced metal and oxidant tolerance to plants.
Subject(s)
Alkalies/pharmacology , Chenopodiaceae/metabolism , Metallothionein/metabolism , Plant Proteins/metabolism , Plants, Genetically Modified/metabolism , Salt-Tolerant Plants/metabolism , Stress, Physiological , Arabidopsis/drug effects , Arabidopsis/genetics , Arabidopsis/metabolism , Cadmium/pharmacology , Chenopodiaceae/drug effects , Chenopodiaceae/genetics , Cloning, Molecular , Gene Expression Regulation, Plant , Hydrogen Peroxide/pharmacology , Metallothionein/genetics , Plant Proteins/genetics , Plants, Genetically Modified/drug effects , Plants, Genetically Modified/genetics , Saccharomyces cerevisiae/drug effects , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Salt-Tolerant Plants/drug effects , Salt-Tolerant Plants/genetics , Sodium/pharmacologyABSTRACT
Jing Wan Hong ointment contains 30 kinds of Chinese herbs, with functions of activating blood circulation to disperse blood stasis, clearing heat, eliminating dampness, and reducing swelling by detoxification. Therefore, Jing Wan Hong ointment may facilitate the healing of ulcers. The aim of this study was to evaluate the efficacy and mechanisms of Jing Wan Hong ointment for healing diabetic foot ulceration in Wistar rats induced by streptozotocin and sciatic nerve damage. The results showed that Jing Wan Hong ointment had a marked effect on foot ulcers in diabetic rats induced by initial nerve injury. These effects were manifested by reducing the foot ulcer size and Wagner grade after seven days of treatment. The diabetic rats with foot ulcers were almost healed after 21 days of treatment. Moreover, the mechanisms of this effect seem to be dependent on increased expression of PDGF mRNA, but there was no influence on the expression of TGF-ß, VEGF, and FLT-1 mRNA.
Subject(s)
Diabetic Foot/drug therapy , Drugs, Chinese Herbal/administration & dosage , Wound Healing/drug effects , Administration, Cutaneous , Animals , Diabetes Mellitus, Experimental/chemically induced , Diabetes Mellitus, Experimental/complications , Diabetic Foot/diagnosis , Diabetic Foot/etiology , Diabetic Foot/genetics , Diabetic Foot/metabolism , Male , Ointments , Platelet-Derived Growth Factor/genetics , Platelet-Derived Growth Factor/metabolism , RNA, Messenger/metabolism , Rats, Wistar , Sciatic Neuropathy/complications , Streptozocin , Time Factors , Up-RegulationABSTRACT
Metallothioneins (MTs) are cysteine-rich proteins of low molecular weight with many attributed functions, such as providing protection against metal toxicity, being involved in regulation of metal ions uptake that can impact plant physiology and providing protection against oxidative stress. However, the precise function of the metallothionein-like proteins such as the one coded for rgMT gene isolated from rice (Oryza sativa L.) is not completely understood. The whole genome analysis of rice (O. sativa) showed that the rgMT gene is homologue to the Os11g47809 on chromosome 11 of O. sativa sp. japonica genome. This study used the rgMT coding sequence to create transgenic lines to investigate the subcellular localization of the protein, as well as the impact of gene expression in yeast (Saccharomyces cerevisiae) and Arabidopsis thaliana under heavy metal ion, salt and oxidative stresses. The results indicate that the rgMT gene was expressed in the cytoplasm of transgenic cells. Yeast cells transgenic for rgMT showed vigorous growth compared to the nontransgenic controls when exposed to 7 mM CuCl2, 10 mM FeCl2, 1 M NaCl, 24 mM NaHCO3 and 3.2 mM H2O2, but there was no significant difference for other stresses tested. Similarly, Arabidopsis transgenic for rgMT displayed significantly improved seed germination rates over that of the control when the seeds were stressed with 100 µM CuCl2 or 1 mM H2O2. Increased biomass was observed in the presence of 100 µM CuCl2, 220 µM FeCl2, 3 mM Na2CO3, 5 mM NaHCO3 or 1 mM H2O2. These results indicate that the expression of the rice rgMT gene in transgenic yeast and Arabidopsis is implicated in improving their tolerance for certain salt and peroxide stressors.
Subject(s)
Metallothionein/biosynthesis , Oryza/genetics , Oxidative Stress/genetics , Arabidopsis/genetics , Gene Expression Regulation, Plant/drug effects , Hydrogen Peroxide/pharmacology , Metallothionein/genetics , Plants, Genetically Modified , Saccharomyces cerevisiae/genetics , Salts/pharmacologyABSTRACT
High abundance proteins like ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco) impose a consistent challenge for the whole proteome characterization using shot-gun proteomics. To address this challenge, we developed and evaluated Polyethyleneimine Assisted Rubisco Cleanup (PARC) as a new method by combining both abundant protein removal and fractionation. The new approach was applied to a plant insect interaction study to validate the platform and investigate mechanisms for plant defense against herbivorous insects. Our results indicated that PARC can effectively remove Rubisco, improve the protein identification, and discover almost three times more differentially regulated proteins. The significantly enhanced shot-gun proteomics performance was translated into in-depth proteomic and molecular mechanisms for plant insect interaction, where carbon re-distribution was used to play an essential role. Moreover, the transcriptomic validation also confirmed the reliability of PARC analysis. Finally, functional studies were carried out for two differentially regulated genes as revealed by PARC analysis. Insect resistance was induced by over-expressing either jacalin-like or cupin-like genes in rice. The results further highlighted that PARC can serve as an effective strategy for proteomics analysis and gene discovery.
Subject(s)
Genomics/methods , Plant Proteins/isolation & purification , Plants/genetics , Plants/metabolism , Proteomics/methods , Ribulose-Bisphosphate Carboxylase/isolation & purification , Animals , Genome, Plant , Host-Pathogen Interactions/genetics , Oryza/genetics , Oryza/metabolism , Oryza/parasitology , Plant Diseases/genetics , Plant Proteins/genetics , Plant Proteins/metabolism , Plants/parasitology , Plants, Genetically Modified , Spodoptera/pathogenicityABSTRACT
miR-146b-5p is frequently down-regulated in solid tumours, including prostate cancer, pancreatic cancer, and glioblastoma. However, the tumour-suppressive effects of miR-146b-5p in malignant gliomas have not been investigated thoroughly. Here, we found that decreased miR-146b-5p expression was strongly correlated with chromosome 10q loss in gliomas, especially glioblastomas. The overexpression of miR-146b-5p in glioblastoma cell lines led to MMP16 mRNA silencing, MMP2 inactivation, and the inhibition of tumour cell migration and invasion. Our results suggest that the restoration of miR-146b-5p expression may be a feasible approach for inhibiting the migration and invasion of malignant gliomas.
Subject(s)
Cell Movement/genetics , Glioma/genetics , Glioma/pathology , Matrix Metalloproteinase 16/genetics , MicroRNAs/genetics , Base Sequence , Cell Line, Tumor , Chromosome Deletion , Chromosomes, Human, Pair 10 , Gelatinases/metabolism , Gene Expression , Gene Knockdown Techniques , Humans , Matrix Metalloproteinase 16/chemistry , Matrix Metalloproteinase 16/metabolism , MicroRNAs/chemistry , MicroRNAs/metabolism , Neoplasm InvasivenessABSTRACT
PURPOSE: To compare the clinical effects of miniscrews and conventional utility arch in the maxillary incisors intrusion. METHODS: Twenty adult patients with III degree deep bite were chosen and divided into two groups randomly. The cephalometric films, intraoral periapical radiographies with metal guide bars, and periodontium parameters were taken before and after intrusion. The major measurement items included alteration of the maxillary first molar in vertical dimension, torque and length of maxillary central incisors, overbite, clinical crown of maxillary central incisors, probing depth (PD), gingival index (GI), plaque index (PI) and etc. PASW statistics 18.0 was used for statistical analysis. RESULTS: In the miniscrew group, there was no change of U1-SN, U6-PP distance, U6-SN(P<0.05), but significant reductions in overbite and U1-Stm distance(P<0.05). The probing depth (PD) increased(P<0.05), and the width of keratinized gingiva(WKG) decreased significantly(P<0.05). In the conventional utility arch group, significant differences were found in U1-SN, U6-SN, OP-SN, PLI2(P<0.05). There was no difference in periodontal supporting tissue changes between the two groups(P>0.05). CONCLUSIONS: Compared with the conventional utility arch, minisrew has advantages on intruding upper incisors, and improving lip-tooth relationships. Less treatment period is needed in the minisrew group, and the impact on periodontal supporting tissue is similar.
Subject(s)
Incisor , Orthodontic Anchorage Procedures , Adult , Cephalometry , Humans , Molar , Periodontal Index , Tooth Movement TechniquesABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Dipsacus asper Wall (Dipsacaceae), Salvia miltiorrhiza (Salvia) and Drynaria fortunei (Drynaria) have been traditionally used in Chinese medicine as the main ingredient of many formulations for the treatment of cardiovascular and inflammatory diseases. AIM OF THE STUDY: This study was aimed to evaluate the effect of the traditional Chinese medicine (TCM) Dipsacaceae, Salvia and Drynaria on bone remodeling during orthodontic tooth movement (OTM). SETTING: This study was conducted in School of Stomatology, Shandong University between 2009 and 2010 [Jinan, Shandong, China]. MATERIALS AND METHODS: Ninety-six eight-week-old female SPF Wistar rats 180-200 g were selected and randomly divided into four groups of 24: Dipsacaceae group, Salvia group, Drynaria group and control group. Animal models for orthodontic tooth movement were then established which consisted of a closed coil spring ligated to the upper first molar and incisors, exerting a force of 40 g during the experimental period. Rats in the TCM groups were given Dipsacaceae, Salvia and Drynaria decoction respectively by intragastric administration 6 g/kg/day and the control group were given normal saline 3 ml. The rats were sacrificed in batch on the 7th, 14th, 21st and 28th days after orthodontic treatment. Slices from periodontium of the upper first molar were observed under optical microscope. Neovascularization, new bone formation and osteoclast number were observed. RESULTS: The upper first molars were drawn mesial by the force. Telangiectasia and new bone formation in periodontal tissue were significantly in the TCM groups compared with the control group. Application of orthodontic forces in the experimental teeth showed a significant increase (P<0.05) of osteoclast number in the TCM group when compared with the control group. In addition, the number of osteoclast had no significant differences among the TCM groups (P>0.05). Osteoclast number in the TCM group and the control group were 10.12±0.058, 10.13±0.022, 10.09±0.047 and 9.55±0.045, respectively. CONCLUSIONS: These findings suggest that the TCM decoction are beneficial to the alveolar bone remodeling by promoting osteoclast differentiation during OTM.