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1.
PLoS One ; 17(7): e0262517, 2022.
Article in English | MEDLINE | ID: mdl-35834536

ABSTRACT

Photosynthetic bacteria are beneficial to plants, but knowledge of photosynthetic bacterial community dynamics in field crops during different growth stages is scarce. The factors controlling the changes in the photosynthetic bacterial community during plant growth require further investigation. In this study, 35 microbial community samples were collected from the seedling, flowering, and mature stages of tomato, cucumber, and soybean plants. 35 microbial community samples were assessed using Illumina sequencing of the photosynthetic reaction center subunit M (pufM) gene. The results revealed significant alpha diversity and community structure differences among the three crops at the different growth stages. Proteobacteria was the dominant bacterial phylum, and Methylobacterium, Roseateles, and Thiorhodococcus were the dominant genera at all growth stages. PCoA revealed clear differences in the structure of the microbial populations isolated from leaf samples collected from different crops at different growth stages. In addition, a dissimilarity test revealed significant differences in the photosynthetic bacterial community among crops and growth stages (P<0.05). The photosynthetic bacterial communities changed during crop growth. OTUs assigned to Methylobacterium were present in varying abundances among different sample types, which we speculated was related to the function of different Methylobacterium species in promoting plant growth development and enhancing plant photosynthetic efficiency. In conclusion, the dynamics observed in this study provide new research ideas for the detailed assessments of the relationship between photosynthetic bacteria and different growth stages of plants.


Subject(s)
Metagenomics , Microbiota , Bacteria , Crops, Agricultural , High-Throughput Nucleotide Sequencing , Metagenome , Microbiota/genetics , Soil Microbiology
2.
J Cell Biochem ; 120(5): 8483-8491, 2019 05.
Article in English | MEDLINE | ID: mdl-30556156

ABSTRACT

BACKGROUND: Diabetic chronic kidney disease (CKD) has become the main cause of death in diabetic patients, but its pathogenesis has not yet been clear. OBJECTIVE: To investigate the effects of reduced glutathione (GSH) on oxidative stress (OS), angiogenesis factors and lymphocyte subsets in diabetic CKD patients. METHODS: A total of 130 subjects were retrospectively studied. The subjects were divided into the control group (45 cases), treatment group (45 cases, treated with reduced GSH), and a healthy control group (40 cases). The levels of superoxide dismutase (SOD), advanced oxidation protein products (AOPP), malondialdehyde (MDA), endostatin (ES), and vascular endothelial growth factor (VEGF), and the percentages of lymphocyte subsets were detected. RESULTS: After treatment, the indexes of OS and angiogenesis and the percentage of CD3- CD19+ B cells were obviously decreased, and the percentages of T cell subsets and natural killer (NK) cell subsets were markedly increased in the treatment group compared with the control group. AOPP was positively correlated with angiogenesis indexes, MDA and CD3- CD19+ B cells, and negatively correlated with SOD and other lymphocyte subsets. SOD was inversely associated with angiogenesis indexes and MDA, and positively associated with lymphocyte subsets. Moreover, MDA had a positive correlation with angiogenesis indexes, B and T cell subsets, and a negative correlation with NK cell subsets. AOPP, MDA, SOD, VEGF, CD3+ T cells, CD3+ CD8+ T cells, CD3- CDl6+ CD56+ NK cells, and CD3- CDl6+ CD56+ NK T cells were the risk factors of diabetic CKD. CONCLUSION: GSH could inhibit OS and abnormal angiogenesis, and improve cellular immune response in CKD patients.


Subject(s)
Diabetic Nephropathies/drug therapy , Glutathione/therapeutic use , Renal Insufficiency, Chronic/drug therapy , Case-Control Studies , Diabetic Nephropathies/pathology , Female , Humans , Lymphocyte Subsets/metabolism , Male , Middle Aged , Neovascularization, Physiologic , Oxidative Stress , Renal Insufficiency, Chronic/pathology , Risk Factors
3.
Behav Brain Funct ; 14(1): 12, 2018 Jun 08.
Article in English | MEDLINE | ID: mdl-29884193

ABSTRACT

BACKGROUND: Predatory stress as a psychological stressor can elicit the activation of the hypothalamic-pituitary-adrenal (HPA) axis, which is involved in the dialogue of the neuroimmunoendocrine network. The brain has been proven to regulate the activity of the HPA axis by way of lateralization. In the present study, we probed the pivotal elements of the HPA circuitry including CRH, GR and a multifunctional cytokine in behavior-lateralized mice to determine their changes when the animals were subjected to predator exposure. METHODS: Behavior-lateralized mice were classified into left-pawed and right-pawed mice through a paw-preference test. Thereafter, mice in the acute stress group received a single 60-min cat exposure, and mice in the chronic group received daily 60-min cat exposure for 14 consecutive days. The plasma CS and TNF-α were determined by ELISA, the hypothalamic CRH mRNA and hippocampal GR mRNA were detected by real-time PCR, and the hippocampal GR protein was detected by western blot analysis. RESULTS: The results revealed that the levels of plasma CS were significantly elevated after chronic predatory exposure in both right-pawed and left-pawed mice; the right-pawed mice exhibited a higher plasma CS level than the left-pawed mice. Similarly, the acute or chronic cat exposure could induce the release of plasma TNF-α, and the left-pawed mice tended to show a higher level after the acute stress. Chronic stress significantly upregulated the expression of hypothalamic CRH mRNA in both left-pawed and right-pawed mice. Normally, the left-pawed mice exhibited a higher GR expression in the hippocampus than the right-pawed mice. After the cat exposure, the expression of GR in both left-pawed and right-pawed mice was revealed to be greatly downregulated. CONCLUSION: Our findings indicate that predatory stress can invoke a differential response of stressful elements in behavior-lateralized mice. Some of these responses shaped by behavioral lateralization might be helpful for facilitating adaption to various stimuli.


Subject(s)
Functional Laterality/physiology , Hypothalamo-Hypophyseal System/metabolism , Pituitary-Adrenal System/metabolism , Predatory Behavior/physiology , Stress, Psychological/blood , Stress, Psychological/psychology , Animals , Cats , Female , Male , Mice , Mice, Inbred BALB C
4.
Int J Mol Sci ; 19(4)2018 Mar 23.
Article in English | MEDLINE | ID: mdl-29570670

ABSTRACT

Oxymatrine (OMT) is a strong immunosuppressive agent that has been used in the clinic for many years. In the present study, by using plaque inhibition, luciferase reporter plasmids, qRT-PCR, western blotting, and ELISA assays, we have investigated the effect and mechanism of OMT on influenza A virus (IAV) replication and IAV-induced inflammation in vitro and in vivo. The results showed that OMT had excellent anti-IAV activity on eight IAV strains in vitro. OMT could significantly decrease the promoter activity of TLR3, TLR4, TLR7, MyD88, and TRAF6 genes, inhibit IAV-induced activations of Akt, ERK1/2, p38 MAPK, and NF-κB pathways, and suppress the expressions of inflammatory cytokines and MMP-2/-9. Activators of TLR4, p38 MAPK and NF-κB pathways could significantly antagonize the anti-IAV activity of OMT in vitro, including IAV replication and IAV-induced cytopathogenic effect (CPE). Furthermore, OMT could reduce the loss of body weight, significantly increase the survival rate of IAV-infected mice, decrease the lung index, pulmonary inflammation and lung viral titter, and improve pulmonary histopathological changes. In conclusion, OMT possesses anti-IAV and anti-inflammatory activities, the mechanism of action may be linked to its ability to inhibit IAV-induced activations of TLR4, p38 MAPK, and NF-κB pathways.


Subject(s)
Alkaloids/pharmacology , Influenza A virus/drug effects , NF-kappa B/metabolism , Quinolizines/pharmacology , Toll-Like Receptor 4/metabolism , p38 Mitogen-Activated Protein Kinases/metabolism , A549 Cells , Animals , Antiviral Agents/pharmacology , Cell Line , DNA Replication/drug effects , Dogs , Humans
5.
PLoS One ; 13(1): e0191793, 2018.
Article in English | MEDLINE | ID: mdl-29385192

ABSTRACT

Rhein, an anthraquinone compound existing in many traditional herbal medicines, has anti-inflammatory, antioxidant, antitumor, antiviral, hepatoprotective, and nephroprotective activities, but its anti-influenza A virus (IAV) activity is ambiguous. In the present study, through plaque inhibition assay, time-of-addition assay, antioxidant assay, qRT-PCR, ELISA, and western blotting assays, we investigated the anti-IAV effect and mechanism of action of rhein in vitro and in vivo. The results showed that rhein could significantly inhibit IAV adsorption and replication, decrease IAV-induced oxidative stress, activations of TLR4, Akt, p38, JNK MAPK, and NF-κB pathways, and production of inflammatory cytokines and matrix metalloproteinases in vitro. Oxidant H2O2 and agonists of TLR4, Akt, p38/JNK and IKK/NF-κB could significantly antagonize the inhibitory effects of rhein on IAV-induced cytopathic effect (CPE) and IAV replication. Through an in vivo test in mice, we also found that rhein could significantly improve the survival rate, lung index, pulmonary cytokines, and pulmonary histopathological changes. Rhein also significantly decreased pulmonary viral load at a high dose. In conclusion, rhein can inhibit IAV adsorption and replication, and the mechanism of action to inhibit IAV replication may be due to its ability to suppress IAV-induced oxidative stress and activations of TLR4, Akt, p38, JNK MAPK, and NF-κB signal pathways.


Subject(s)
Anthraquinones/pharmacology , Antiviral Agents/pharmacology , Influenza A Virus, H1N1 Subtype/drug effects , A549 Cells , Animals , Cytokines/biosynthesis , Dogs , Female , Humans , Influenza A Virus, H1N1 Subtype/pathogenicity , Influenza A Virus, H1N1 Subtype/physiology , Lung/drug effects , Lung/pathology , MAP Kinase Signaling System/drug effects , Madin Darby Canine Kidney Cells , Male , Matrix Metalloproteinases/biosynthesis , Mice , Mice, Inbred C57BL , NF-kappa B/metabolism , Orthomyxoviridae Infections/drug therapy , Orthomyxoviridae Infections/metabolism , Orthomyxoviridae Infections/virology , Oxidative Stress/drug effects , Pneumonia, Viral/drug therapy , Pneumonia, Viral/pathology , Pneumonia, Viral/physiopathology , Proto-Oncogene Proteins c-akt/metabolism , Signal Transduction/drug effects , Toll-Like Receptor 4/metabolism , Virus Attachment/drug effects , Virus Replication/drug effects
6.
Molecules ; 22(10)2017 Oct 18.
Article in English | MEDLINE | ID: mdl-29057806

ABSTRACT

Lasting activations of toll-like receptors (TLRs), MAPK and NF-κB pathways can support influenza A virus (IAV) infection and promote pneumonia. In this study, we have investigated the effect and mechanism of action of emodin on IAV infection using qRT-PCR, western blotting, ELISA, Nrf2 luciferase reporter, siRNA and plaque inhibition assays. The results showed that emodin could significantly inhibit IAV (ST169, H1N1) replication, reduce IAV-induced expressions of TLR2/3/4/7, MyD88 and TRAF6, decrease IAV-induced phosphorylations of p38/JNK MAPK and nuclear translocation of NF-κB p65. Emodin also activated the Nrf2 pathway, decreased ROS levels, increased GSH levelss and GSH/GSSG ratio, and upregulated the activities of SOD, GR, CAT and GSH-Px after IAV infection. Suppression of Nrf2 via siRNA markedly blocked the inhibitory effects of emodin on IAV-induced activations of TLR4, p38/JNK, and NF-κB pathways and on IAV-induced production of IL-1ß, IL-6 and expression of IAV M2 protein. Emodin also dramatically increased the survival rate of mice, reduced lung edema, pulmonary viral titer and inflammatory cytokines, and improved lung histopathological changes. In conclusion, emodin can inhibit IAV replication and influenza viral pneumonia, at least in part, by activating Nrf2 signaling and inhibiting IAV-induced activations of the TLR4, p38/JNK MAPK and NF-κB pathways.


Subject(s)
Emodin/administration & dosage , Influenza A virus/drug effects , Influenza, Human/drug therapy , Pneumonia/drug therapy , Animals , Disease Models, Animal , Humans , Influenza A virus/genetics , Influenza A virus/pathogenicity , Influenza, Human/complications , Influenza, Human/genetics , Influenza, Human/virology , Mice , Myeloid Differentiation Factor 88/genetics , NF-E2-Related Factor 2/genetics , Pneumonia/etiology , Pneumonia/pathology , Pneumonia/virology , RNA, Small Interfering/administration & dosage , Signal Transduction/drug effects , TNF Receptor-Associated Factor 6/genetics , Toll-Like Receptor 4/genetics , Transcription Factor RelA/genetics , Virus Replication/drug effects , p38 Mitogen-Activated Protein Kinases/genetics
7.
Psychiatry Res Neuroimaging ; 265: 72-76, 2017 Jul 30.
Article in English | MEDLINE | ID: mdl-28017479

ABSTRACT

White matter degradation is a major part of the pathogenesis of Alzheimer's disease (AD). The fornix is the predominant outflow tract from the hippocampus, and alterations to its microstructure in patients with AD are still being explored. Diffusion tensor imaging (DTI) is an in vivo neuroimaging technique that can provide unique information about alterations in tissue microstructure, which can indicate underlying neurobiological process at the microstructural level. In this prospective study, DTI was used to assess and analyze the microstructural features of the fornix in subjects with AD (n = 17), mild cognitive impairment (MCI; n = 12) and healthy controls (n = 17). DTI was performed using Explore DTI software and the FSL package. Within the fornix, patients with AD showed decreased fractional anisotropy values and length of fiber tracts of the fornix relative to healthy controls, but higher mean diffusivity values. MCI subjects showed a trend towards elevated mean diffusivity values in the fornix. The data suggest that DTI provides supporting information on the microstructural alteration of the fornix in patients with AD, and that these diffusion characteristics of the fornix may be helpful for the clinical diagnosis of AD.


Subject(s)
Alzheimer Disease/diagnostic imaging , Cognitive Dysfunction/diagnostic imaging , Diffusion Tensor Imaging/methods , Fornix, Brain/diagnostic imaging , Adult , Aged , Aged, 80 and over , Alzheimer Disease/diagnosis , Anisotropy , Cognitive Dysfunction/diagnosis , Female , Fornix, Brain/pathology , Humans , Male , Middle Aged , Prospective Studies , Reproducibility of Results , Sensitivity and Specificity
8.
Medicine (Baltimore) ; 95(52): e5543, 2016 Dec.
Article in English | MEDLINE | ID: mdl-28033250

ABSTRACT

RATIONALE: Germinomas are sensitive to radiation therapy and chemotherapy; therefore, correct imaging diagnosis is crucial for them. However, the imaging findings of germinomas originating from off-midline regions displayed different patterns from those originating from midline areas. PATIENT CONCERNS: The objective of this study is to describe the radiologic features of primary ectopic germinoma. We reviewed the MR and CT findings of 12 patients with histologically proven off-midline ectopic germinomas with off-midline locations. INTERVENTIONS: All of these patients underwent conventional MR images and 3 of them underwent diffusion images. Additional CT images were available in 3 patients. Analysis was focused on the shape and entity of tumors in images, signs of hemiatrophy, and the involvement of fibers in diffusion images. OUTCOMES: Well-defined (8/12) and ill-defined margin masses (4/12) were identified according to the shape of the mass. Multicystic masses were seen in 11 of the 12 patients. The solid component of the tumors had a high density (3/3) with calcifications (2/3) on CT images, iso- to hypointensity in T2WI (11/12) and restricted diffusion on apparent diffusion coefficient (ADC) maps (3/3). Hemiatrophy was observed in 5 cases and progressive hemiatrophy was observed in 1 case. Other signs included mild peritumoral edema (10/12), and hydrocephalus (7/12). Additionally, infiltration of the corticospinal tract (CST) was identified on diffusion tensor imaging (DTI) (2/2). LESSONS: The results indicate that multicysitic entities and hypointensities in solid components on T2WI and hemiatrophy are the imaging features of ectopic germinomas. DTI has potential for assessing CST involvement.


Subject(s)
Brain Neoplasms/diagnostic imaging , Germinoma/diagnostic imaging , Magnetic Resonance Imaging , Tomography, X-Ray Computed , Adolescent , Adult , Brain Edema/diagnostic imaging , Brain Neoplasms/pathology , Calcinosis/diagnostic imaging , Child , Female , Germinoma/pathology , Humans , Hydrocephalus/diagnostic imaging , Magnetic Resonance Imaging/methods , Male , Young Adult
9.
Molecules ; 20(4): 6794-807, 2015 Apr 15.
Article in English | MEDLINE | ID: mdl-25884554

ABSTRACT

Salvia miltiorrhiza Bunge has been reported to possess excellent antifibrotic activity. In this study, we have investigated the effect and mechanism of tanshinone IIA (Tan-IIA), salvianolic acid A (Sal-A) and salvianolic acid B (Sal-B), the important active compounds of Salvia miltiorrhiza Bunge, on areca nut extract (ANE)-induced oral submucous fibrosis (OSF) in vitro. Through human procollagen gene promoter luciferase reporter plasmid assay, hydroxyproline assay, gelatin zymography assay, qRT-PCR, ELISA and Western blot assay, the influence of these three compounds on ANE-stimulated cell viability, collagen accumulation, procollagen gene transcription, MMP-2/-9 activity, MMP-1/-13 and TIMP-1/-2 expression, cytokine secretion and the activation of PI3K/AKT, ERK/JNK/p38 MAPK and TGF-ß/Smads pathways were detected. The results showed that Tan-IIA, Sal-A and Sal-B could significantly inhibit the ANE-stimulated abnormal viability and collagen accumulation of mice oral mucosal fibroblasts (MOMFs), inhibit the transcription of procollagen gene COL1A1 and COL3A1, increase MMP-2/-9 activity, decrease TIMP-1/-2 expression and inhibit the transcription and release of CTGF, TGF-ß1, IL-6 and TNF-α; Tan-IIA, Sal-A and Sal-B also inhibited the ANE-induced activation of AKT and ERK MAPK pathways in MOMFs and the activation of TGF-ß/Smads pathway in HaCaT cells. In conclusion, Tan-IIA, Sal-A and Sal-B possess excellent antifibrotic activity in vitro and can possibly be used to promote the rehabilitation of OSF patients.


Subject(s)
Abietanes/pharmacology , Areca/chemistry , Benzofurans/pharmacology , Caffeic Acids/pharmacology , Lactates/pharmacology , Nuts/chemistry , Oral Submucous Fibrosis/etiology , Plant Exudates/adverse effects , Animals , Cell Line , Cell Survival/drug effects , Collagen/genetics , Collagen/metabolism , Enzyme Activation/drug effects , Extracellular Signal-Regulated MAP Kinases/metabolism , Fibroblasts/drug effects , Fibroblasts/metabolism , Gene Expression Regulation/drug effects , Humans , In Vitro Techniques , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Mice , Oral Submucous Fibrosis/drug therapy , Oral Submucous Fibrosis/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Signal Transduction/drug effects , Smad Proteins/metabolism , Tissue Inhibitor of Metalloproteinase-1/genetics , Tissue Inhibitor of Metalloproteinase-1/metabolism , Tissue Inhibitor of Metalloproteinase-2/genetics , Tissue Inhibitor of Metalloproteinase-2/metabolism , Transforming Growth Factor beta/biosynthesis
10.
J Oral Pathol Med ; 43(6): 464-70, 2014 Jul.
Article in English | MEDLINE | ID: mdl-24484214

ABSTRACT

BACKGROUND: Oral submucous fibrosis (OSF) is a premalignant and fibrosing disease, which is closely associated with the habit of chewing areca nut. Panax notoginseng Buck F. H. Chen is an often used antifibrotic and antitumor agent. To treat areca nut-induced OSF, we have developed a chewable tablet, in which one of the major medicines is total Panax notoginseng saponins (PNS). In this study, we have investigated the antifibrotic effect and mechanism of PNS on areca nut-induced OSF in vitro. METHODS: Through human procollagen gene promoter luciferase reporter plasmid, hydroxyproline assay, gelatin zymography, qRT-PCR, ELISA, and Western blot, the influences of PNS on areca nut extract (ANE)-induced cell growth, collagen accumulation, procollagen gene transcription, MMP-2/-9 activity, MMP-1/-13 and TIMP-1/-2 expression, cytokine secretion, and the activation of PI3K/AKT, ERK/JNK/p38 MAPK, and TGFß/Smads pathways were detected. RESULTS: Panax notoginseng saponins could inhibit the ANE-induced abnormal growth and collagen accumulation of oral mucosal fibroblasts in a concentration-dependent manner. PNS (25 µg/ml) could significantly inhibit the ANE-induced expression of Col1A1 and Col3A1, augment the ANE-induced decrease of MMP-2/-9 activity, inhibit the ANE-induced increase of TIMP-1/-2 expression, and decrease the ANE-induced transcription and release of CTGF, TGFß1, IL-6, and TNFα. PNS (25 µg/ml) also significantly inhibited the ANE-induced activation of AKT and ERK/JNK/p38 MAPK pathways in oral mucosal fibroblasts and the ANE-induced activation of TGFß/smad pathway in HaCaT cells. CONCLUSION: Panax notoginseng saponins possess excellent anti-OSF activity, and its mechanism may be related to its ability to inhibit the ANE-induced activation of PI3K/AKT, ERK/JNK/p38 MAPK, and TGFß/smad pathways.


Subject(s)
Areca/adverse effects , Mouth Mucosa/drug effects , Nuts/adverse effects , Oral Submucous Fibrosis/pathology , Panax notoginseng , Plant Extracts/pharmacology , Saponins/pharmacology , Animals , Cell Culture Techniques , Cell Line , Collagen Type I/drug effects , Collagen Type I, alpha 1 Chain , Collagen Type III/drug effects , Connective Tissue Growth Factor/drug effects , Fibroblasts/drug effects , Humans , Hydroxyproline/analysis , Interleukin-6/analysis , MAP Kinase Signaling System/drug effects , Matrix Metalloproteinase 2/drug effects , Matrix Metalloproteinase 9/drug effects , Mice , Mice, Inbred BALB C , Mouth Mucosa/cytology , Oral Submucous Fibrosis/etiology , Phosphatidylinositol 3-Kinases/drug effects , Plant Extracts/adverse effects , Proto-Oncogene Proteins c-akt/drug effects , Smad Proteins/drug effects , Tissue Inhibitor of Metalloproteinase-1/drug effects , Tissue Inhibitor of Metalloproteinase-2/drug effects , Transforming Growth Factor beta1/drug effects , Tumor Necrosis Factor-alpha/drug effects
11.
Asian Pac J Cancer Prev ; 15(24): 10893-8, 2014.
Article in English | MEDLINE | ID: mdl-25605197

ABSTRACT

Several molecular markers have been proposed as predictors of outcome in patients with glioblastomas. We investigated the prognostic significance of O6-methylguanine-DNA methyltransferase (MGMT) promoter methylation and TP53 mutation status dependent on isocitrate dehydrogenase 1 (IDH1) mutation in glioblastoma patients. A cohort of 78 patients with histologically confirmed glioblastomas treated with radiation therapy and chemotherapy were reviewed retrospectively. We evaluated the prognostic value of MGMT promoter methylation and TP53 mutation status with regard to progression-free survival (PFS) and overall survival (OS). It was revealed that mutations in IDH1, promoter methylation of MGMT, TP53 mutation, age, Karnofsky performance status (KFS), and extension of resection were independent prognostic factors. In patients with an IDH1 mutation, those with an MGMT methylation were associated with longer PFS (p=0.016) and OS (p=0.013). Nevertheless, the presence of TP53 mutation could stratify the PFS and OS of patients with IDH1 wild type (p=0.003 and 0.029 respectively, log-rank). The MGMT promoter methylation and TP53 mutation were associated with a favorable outcome of patients with and without mutant IDH1, respectively. The results indicate that glioblastomas with MGMT methylation or TP53 mutations have improved survival that may be influenced by IDH1 mutation status.


Subject(s)
DNA Methylation , DNA Modification Methylases/genetics , DNA Repair Enzymes/genetics , Glioblastoma/genetics , Isocitrate Dehydrogenase/genetics , Mutation/genetics , Promoter Regions, Genetic/genetics , Tumor Suppressor Protein p53/genetics , Tumor Suppressor Proteins/genetics , Adult , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Biomarkers, Tumor/genetics , Brain Neoplasms/genetics , Brain Neoplasms/mortality , Brain Neoplasms/pathology , Brain Neoplasms/therapy , Chemoradiotherapy , Female , Follow-Up Studies , Glioblastoma/mortality , Glioblastoma/pathology , Glioblastoma/therapy , Humans , Male , Middle Aged , Neoplasm Staging , Prognosis , Retrospective Studies , Survival Rate , Young Adult
12.
Antimicrob Agents Chemother ; 57(9): 4433-43, 2013 Sep.
Article in English | MEDLINE | ID: mdl-23836164

ABSTRACT

It has been reported that autophagy is involved in the replication of many viruses. In this study, we screened 89 medicinal plants, using an assay based on the inhibition of the formation of the Atg12-Atg5/Atg16 heterotrimer, an important regulator of autophagy, and selected Silybum marianum L. for further study. An antiviral assay indicated that silybin (S0), the major active compound of S. marianum L., can inhibit influenza A virus (IAV) infection. We later synthesized 5 silybin derivatives (S1 through S5) and found that 23-(S)-2-amino-3-phenylpropanoyl-silybin (S3) had the best activity. When we compared the polarities of the substituent groups, we found that the hydrophobicity of the substituent groups was positively correlated with their activities. We further studied the mechanisms of action of these compounds and determined that S0 and S3 also inhibited both the formation of the Atg12-Atg5/Atg16 heterotrimer and the elevated autophagy induced by IAV infection. In addition, we found that S0 and S3 could inhibit several components induced by IAV infection, including oxidative stress, the activation of extracellular signal-regulated kinase (ERK)/p38 mitogen-activated protein kinase (MAPK) and IκB kinase (IKK) pathways, and the expression of autophagic genes, especially Atg7 and Atg3. All of these components have been reported to be related to the formation of the Atg12-Atg5/Atg16 heterotrimer, which might validate our screening strategy. Finally, we demonstrated that S3 can significantly reduce influenza virus replication and the associated mortality in infected mice. In conclusion, we identified 23-(S)-2-amino-3-phenylpropanoyl-silybin as a promising inhibitor of IAV infection.


Subject(s)
Antiviral Agents/pharmacology , Influenza A Virus, H1N1 Subtype/drug effects , Plant Extracts/chemistry , Silybum marianum/chemistry , Silymarin/analogs & derivatives , Animals , Antiviral Agents/chemical synthesis , Antiviral Agents/isolation & purification , Autophagy/drug effects , Autophagy-Related Protein 12 , Autophagy-Related Protein 5 , Autophagy-Related Proteins , Carrier Proteins/antagonists & inhibitors , Carrier Proteins/genetics , Carrier Proteins/metabolism , Chlorocebus aethiops , Dogs , Gene Expression Regulation , High-Throughput Screening Assays , Humans , Influenza A Virus, H1N1 Subtype/growth & development , Madin Darby Canine Kidney Cells , Microtubule-Associated Proteins/antagonists & inhibitors , Microtubule-Associated Proteins/genetics , Microtubule-Associated Proteins/metabolism , Mitogen-Activated Protein Kinases/antagonists & inhibitors , Mitogen-Activated Protein Kinases/genetics , Mitogen-Activated Protein Kinases/metabolism , Oxidative Stress/drug effects , Plasmids , Protein Multimerization/drug effects , RNA, Small Interfering/genetics , RNA, Small Interfering/metabolism , Signal Transduction , Silymarin/chemical synthesis , Silymarin/isolation & purification , Silymarin/pharmacology , Small Ubiquitin-Related Modifier Proteins/antagonists & inhibitors , Small Ubiquitin-Related Modifier Proteins/genetics , Small Ubiquitin-Related Modifier Proteins/metabolism , Vero Cells
13.
PLoS One ; 8(4): e61026, 2013.
Article in English | MEDLINE | ID: mdl-23613775

ABSTRACT

Autophagy is involved in many human diseases, such as cancer, cardiovascular disease and virus infection, including human immunodeficiency virus (HIV), hepatitis C virus (HCV), influenza A virus (IAV) and coxsackievirus B3/B4 (CVB3/B4), so a drug screening model targeting autophagy may be very useful for the therapy of these diseases. In our study, we established a drug screening model based on the inhibition of the dissociation of Beclin1-Bcl2 heterodimer, an important negative regulator of autophagy, using bimolecular fluorescence complementation (BiFC) technique for developing novel autophagy inhibitors and anti-IAV agents. From 86 examples of traditional Chinese medicines, we found Syzygium aromaticum L. had the best activity. We then determined the anti-autophagy and anti-IAV activity of eugenol, the major active compound of Syzygium aromaticum L., and explored its mechanism of action. Eugenol could inhibit autophagy and IAV replication, inhibited the activation of ERK, p38MAPK and IKK/NF-κB signal pathways and antagonized the effects of the activators of these pathways. Eugenol also ameliorated the oxidative stress and inhibited the expressions of autophagic genes. We speculated that the mechanism underlying might be that eugenol inhibited the oxidative stress and the activation of ERK1/2, p38MAPK and IKK/NF-κB pathways, subsequently inhibited the dissociation of Beclin1-Bcl2 heterodimer and autophagy, and finally impaired IAV replication. These results might conversely display the reasonableness of the design of our screening model. In conclusion, we have established a drug screening model for developing novel autophagy inhibitor, and find eugenol as a promising inhibitor for autophagy and IAV infection.


Subject(s)
Antiviral Agents/pharmacology , Autophagy/drug effects , Drug Evaluation, Preclinical/methods , Eugenol/pharmacology , Influenza A virus/drug effects , Cell Line , Drugs, Chinese Herbal/pharmacology , Humans , Syzygium/chemistry
14.
PLoS One ; 8(1): e53237, 2013.
Article in English | MEDLINE | ID: mdl-23308170

ABSTRACT

BACKGROUND: Susceptibility weighted imaging (SWI) is a new MRI technique which has been proved very useful in the diagnosis of brain diseases, but few study was performed on its value in prostatic diseases. The aim of the present study was to investigate the value of SWI in distinguishing prostate cancer from benign prostatic hyperplasia and detecting prostatic calcification. METHODOLOGY/PRINCIPAL FINDINGS: 23 patients with prostate cancer and 53 patients with benign prostatic hyperplasia proved by prostate biopsy were scanned on a 3.0T MR and a 16-row CT scanner. High-resolution SWI, conventional MRI and CT were performed on all patients. The MRI and CT findings, especially SWI, were analyzed and compared. The analyses revealed that 19 out of 23 patients with prostate cancer presented hemorrhage within tumor area on SWI. However, in 53 patients with benign prostatic hyperplasia, hemorrhage was detected only in 1 patient in prostate by SWI. When comparing SWI, conventional MRI and CT in detecting prostate cancer hemorrhage, out of the 19 patients with prostate cancer who had prostatic hemorrhage detected by SWI, the prostatic hemorrhage was detected in only 7 patients by using conventional MRI, and none was detected by CT. In addition, CT demonstrated calcifications in 22 patients which were all detected by SWI whereas only 3 were detected by conventional MRI. Compared to CT, SWI showed 100% in the diagnostic sensitivity, specificity, accuracy, positive predictive value(PPV) and negative predictive value(NPV) in detecting calcifications in prostate but conventional MRI demonstrated 13.6% in sensitivity, 100% in specificity, 75% in accuracy, 100% in PPV and 74% in NPV. CONCLUSIONS: More apparent prostate hemorrhages were detected on SWI than on conventional MRI or CT. SWI may provide valuable information for the differential diagnosis between prostate cancer and prostatic hyperplasia. Filtered phase images can identify prostatic calcifications as well as CT.


Subject(s)
Magnetic Resonance Imaging/methods , Prostate/pathology , Prostatic Hyperplasia/diagnosis , Prostatic Neoplasms/diagnosis , Aged , Aged, 80 and over , Humans , Male , Middle Aged , Prospective Studies , Prostatic Hyperplasia/pathology , Prostatic Neoplasms/pathology
15.
PLoS One ; 8(12): e83753, 2013.
Article in English | MEDLINE | ID: mdl-24386270

ABSTRACT

Cardiomyocytes are quite resistant to gene transfer using standard techniques. We developed an expression vector carrying an attenuated but infectious and replicative coxsackievirus B3 (CVB3) genome, and unique ClaI-StuI cloning sites for an exogenous gene, whose product can be released from the nascent viral polyprotein by 2A(pro) cleavage. This vector was tested as an expression vehicle for green fluorescent protein (GFP). The vector transiently expressed GFP in cell cultures for at least ten passages and delivered functional GFP to the infected cardiomyocytes for at least 6 days. Moreover, the recombinant viruses showed virulence attenuation in vitro and in vivo. The findings suggest that the recombinant CVB3 vector could be a useful tool for viral tracking study and delivering exogenous proteins to cardiomyocytes.


Subject(s)
Enterovirus B, Human/genetics , Gene Transfer Techniques , Genetic Vectors/genetics , Animals , Cell Line , Enterovirus B, Human/pathogenicity , Enterovirus Infections/pathology , Enterovirus Infections/virology , Gene Expression , Gene Order , Genome, Viral , Humans , Male , Mice , Myocytes, Cardiac/metabolism , Transduction, Genetic , Transgenes , Virulence
16.
Chin Med J (Engl) ; 125(24): 4334-7, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23253697

ABSTRACT

BACKGROUND: Reliable early prediction response to therapy and time-to-progression (TTP) remain an important goal of high-grade gliomas (HGGs) research. Proton magnetic resonance spectroscopy ((1)H-MRS) has been applied with variable success in clinical application, and we hypothesize that (1)H-MRS in predictive value should perform well as a marker of TTP in patients treated with radiotherapy (RT) after surgery. METHODS: (1)H-MRS was performed before surgery on 25 patients who had undergone resection of HGGs; then the ratios of lipid/creatine (Lip/Cr) and myo-inositol/creatine (mI/Cr) were determined in the solid tumor. RT response was classified as follows: complete resolution (CR), partial response (PR), stable disease (SD), and progressive disease (PD) by comparison of pre-treatment and post-radiotherapy scans. TTP was defined at the time to radiographic progression by MacDonald criteria. Correlation was evaluated between the ratios of Lip/Cr, mI/Cr and treatment response, TTP. The chi-square test and Pearson correlation test were used for data analyses. RESULTS: Multivariate analysis revealed that the prognostic value of spectroscopic variables was independent of age, sex, WHO histologic grade, extent of surgery, and Karnofsky score (KPS). The correlation between the ratios of lipid/Cr and TTP was significant (r = 0.894, P = 0.000), and between the ratios of mI/Cr and TTP was also significant (r = 0.891, P = 0.000). As predicted, RT response correlated significantly with TTP (r = 0.59, P = 0.002): median TTP was 49.9 days for patients with PD compared with 202.7 days for SD, 208.0 days for PR, and 234.5 days for CR. CONCLUSION: The ratios of Lip/Cr and mI/Cr of the solid tumor region before surgery could provide important information in predicting RT response and TTP in patients with HGGs treated by radiation alone after surgery.


Subject(s)
Glioma/radiotherapy , Magnetic Resonance Spectroscopy/methods , Glioma/surgery , Humans , Multivariate Analysis
17.
PLoS One ; 7(8): e42706, 2012.
Article in English | MEDLINE | ID: mdl-22900043

ABSTRACT

In this research, we have established a drug screening method based on the autophagy signal pathway using the bimolecular fluorescence complementation-fluorescence resonance energy transfer (BiFC-FRET) technique to develop novel anti-influenza A virus (IAV) drugs. We selected Evodia rutaecarpa Benth out of 83 examples of traditional Chinese medicine and explored the mechanisms of evodiamine, the major active component of Evodia rutaecarpa Benth, on anti-IAV activity. Our results showed that evodiamine could significantly inhibit IAV replication, as determined by a plaque inhibition assay, an IAV vRNA promoter luciferase reporter assay and the Sulforhodamine B method using cytopathic effect (CPE) reduction. Additionally, evodiamine could significantly inhibit the accumulation of LC3-II and p62, and the dot-like aggregation of EGFP-LC3. This compound also inhibited the formation of the Atg5-Atg12/Atg16 heterotrimer, the expressions of Atg5, Atg7 and Atg12, and the cytokine release of TNF-α, IL-1ß, IL-6 and IL-8 after IAV infection. Evodiamine inhibited IAV-induced autophagy was also dependent on its action on the AMPK/TSC2/mTOR signal pathway. In conclusion, we have established a new drug screening method, and selected evodiamine as a promising anti-IAV compound.


Subject(s)
Antiviral Agents/pharmacology , Autophagy/drug effects , Drug Evaluation, Preclinical/methods , Influenza A virus/drug effects , Quinazolines/pharmacology , Signal Transduction/drug effects , Adenylate Kinase/metabolism , Animals , Autophagy/genetics , Carrier Proteins/genetics , Carrier Proteins/metabolism , Cell Line , Cytokines/biosynthesis , Gene Expression/drug effects , Humans , Microtubule-Associated Proteins/genetics , Microtubule-Associated Proteins/metabolism , Protein Binding/drug effects , Protein Multimerization/drug effects , Small Ubiquitin-Related Modifier Proteins/genetics , Small Ubiquitin-Related Modifier Proteins/metabolism , TOR Serine-Threonine Kinases/metabolism , Tuberous Sclerosis Complex 2 Protein , Tumor Suppressor Proteins/metabolism , Virus Replication/drug effects
18.
Chin Med J (Engl) ; 125(9): 1627-32, 2012 May.
Article in English | MEDLINE | ID: mdl-22800833

ABSTRACT

BACKGROUND: The mechanism of acupuncture analgesia in craniotomy has been widely studied. However, the theoretical basis for selection of acupoints has not been examined. In this study, we used the regional homogeneity method blood oxygen level-dependent (BOLD) signals to determine changes in brain activity in response to transcutaneous electrical stimulation on acupoints and non-acupoints in resting state functional magnetic resonance imaging (fMRI). METHODS: Twelve healthy volunteers were enrolled in this study. BOLD fMRI scanning of the brain was performed for 306 seconds before and 30 minutes after transcutaneous electrical stimulation on acupoints UB63 (Jinmen), LV3 (Tai chong), ST36 (Zusanli), and GB40 (Qiuxu). The procedure was repeated after one week with stimulation on non-acupoints (one was 9 above BL67, the second was 12 above BL67 (Kunlun), the third was 7 above KI3, and the fourth was 10 above KI3 (Taixi)). RESULTS: The regional homogeneity in the acupoint group was increased in the left thalamus, caudate, putamen, lentiform nucleus (BA19, 30, 39), postcentral gyrus, precentral gyrus (BA3, 4, 30, 32), calcarine fissure, middle temporal gyrus (BA30), right superior temporal gyrus, inferior temporal gyrus (BA38), cuneus, and precuneus (BA7, 19) when compared to the non-acupoint group. The regional homogeneity of the acupoint group was decreased in the left cerebellum posterior lobe, middle frontal gyrus (BA10), double-side precuneus (BA7), and the postcentral gyrus (BA40). CONCLUSIONS: The brain region activated following acupoint stimulation is the ipsilateral pain-related brain region, which may relate to the therapeutic effect of acupuncture on pain relief. Further acupoint stimulation causes different central nervous responses compared to non-acupoint stimulation.


Subject(s)
Acupuncture Points , Magnetic Resonance Imaging/methods , Transcutaneous Electric Nerve Stimulation , Acupuncture Therapy , Adult , Female , Humans , Male , Young Adult
19.
Brain Dev ; 34(1): 8-12, 2012 Jan.
Article in English | MEDLINE | ID: mdl-21300489

ABSTRACT

As a study method of resting state network (RSN), resting state functional MRI (rfMRI) can be applied to detect low frequency fluctuate (LFF) in various cerebral areas based on resting state blood oxygen level dependence (BOLD) signals; and it is easier and more consistent than task-related fMRI. The development, features and methods of rfMRI as well as the application in epilepsy were reviewed in this article.


Subject(s)
Brain Mapping/methods , Brain/physiopathology , Epilepsy/physiopathology , Magnetic Resonance Imaging/methods , Humans , Image Interpretation, Computer-Assisted , Rest
20.
Eur J Radiol ; 81(9): 2370-4, 2012 Sep.
Article in English | MEDLINE | ID: mdl-21652157

ABSTRACT

OBJECTIVE: To investigate the added value of volume on post-contrast three dimensional (3D) T1-weighted image (T1WI) over classical cross-sectional area on two dimensional (2D) T1WI in evaluating tumor response in glioblastoma multiforme (GBM). METHODS: Tumor cross-sectional area and volume measurements were performed on 104 MRI studies from 42 adult patients with GBM on post-contrast 5 mm 2D T1WI and isotropic high resolution 3D T1WI, respectively. 52 pairs of MRI scans were analyzed for relative change. Radiographic responses were determined based on change in either area or volume. RESULTS: A high correlation was revealed between tumor size measured by area on thick 2D and volume on high resolution 3D MRI in 104 scans (r=0.82, p<0.001). When four tumor response criteria were used according to the percentage changes (complete response/partial response/stable disease/progression), the kappa coefficient between the area on 2D and volume on 3D was 0.68 (p<0.05) with an overall agreement of 81%. CONCLUSIONS: Tumor cross-sectional area on post-contrast 2D T1WI appears comparable to volume on 3D T1WI and should still be a practical alternate of volume on 3D for evaluating tumor response.


Subject(s)
Brain Neoplasms/pathology , Brain Neoplasms/surgery , Glioblastoma/pathology , Glioblastoma/surgery , Image Interpretation, Computer-Assisted/methods , Imaging, Three-Dimensional/methods , Magnetic Resonance Imaging/methods , Adult , Aged , Aged, 80 and over , Algorithms , Female , Humans , Image Enhancement/methods , Male , Middle Aged , Reproducibility of Results , Sensitivity and Specificity , Treatment Outcome
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