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1.
Exp Ther Med ; 27(5): 221, 2024 May.
Article in English | MEDLINE | ID: mdl-38590563

ABSTRACT

Ischemic cardiomyopathy (ICM) is a serious cardiac disease with a very high mortality rate worldwide, which causes myocardial ischemia and hypoxia as the main damage. Further understanding of the underlying pathological processes of cardiomyocyte injury is key to the development of cardioprotective strategies. Ferroptosis is an iron-dependent form of regulated cell death characterized by the accumulation of lipid hydroperoxides to lethal levels, resulting in oxidative damage to the cell membrane. The current understanding of the role and regulation of ferroptosis in ICM is still limited, especially in the absence of evidence from large-scale transcriptomic data. Through comprehensive bioinformatics analysis of human ICM transcriptome data obtained from the Gene Expression Omnibus database, the present study identified differentially expressed ferroptosis-related genes (DEFRGs) in ICM. Subsequently, their potential biological mechanisms and cross-talk were analyzed, and hub genes were identified by constructing protein-protein interaction networks. Ferroptosis features such as reactive oxygen species generation, changes in ferroptosis marker proteins, iron ion aggregation and lipid oxidation, were identified in the H9c2 anoxic reoxygenation injury model. Finally, the diagnostic ability of Gap junction alpha-1 (GJA1), Solute carrier family 40 member 1 (SLC40A1), Alpha-synuclein (SNCA) were identified through receiver operating characteristic curves and the expression of DEFRGs was verified in an in vitro model. Furthermore, potential drugs (retinoic acid) that could regulate ICM ferroptosis were predicted based on key DEFRGs. The present article presents new insights into the role of ferroptosis in ICM, investigating the regulatory role of ferroptosis in the pathological process of ICM and advocating for ferroptosis as a potential novel therapeutic target for ICM based on evidence from the ICM transcriptome.

3.
J Healthc Eng ; 2021: 2087876, 2021.
Article in English | MEDLINE | ID: mdl-34603642

ABSTRACT

In order to explore the quality management efficiency of applying big data and artificial intelligence in nursing quality index, a method of building a nursing management platform integrating nursing indicators and nursing events is proposed. Based on the investigation of the application demand of nursing information system, the method achieves timely data sharing and transmission through WLAN technology and realizes nursing management monitoring, nursing quality index enquiry, and automatic statistical analysis under the vertical management mode of nursing. The results showed that 77 people (73%) thought the time decreased, 19 people (18%) thought the time was the same, and 9 people (7%) thought the time increased. In terms of intelligent application and big data of nursing information management system, there is a significant difference in nursing management efficiency before and after using nursing management information system (P < 0.001). The nursing management control platform is designed and applied, and the nursing quality control method and actual management process are improved, which is very good for strengthening nursing quality management. The overall optimization of the quality control process is realized, which helps to mobilize the initiative and enthusiasm of nursing staff and continuously improve the effectiveness of nursing management and nursing efficiency.


Subject(s)
Artificial Intelligence , Big Data , Humans , Quality Indicators, Health Care , Technology
4.
Ann Palliat Med ; 10(6): 6687-6693, 2021 Jun.
Article in English | MEDLINE | ID: mdl-34118858

ABSTRACT

BACKGROUND: Henoch-Schonlein purpura (HSP) is a common capillary allergic bleeding disease. To explore the variation of pyroptosis-related inflammatory factors level in the peripheral blood of patients with HSP. METHODS: A total of 87 HSP patients treated in our hospital from June 2020 to March 2021 were selected and divided into the renal impairment group (n=29) and the non-renal impairment group (n=58) according to the presence of hematuria and proteinuria. A total of 50 healthy individuals from the hospital were selected as the control group. The renal impairment and non-renal impairment groups were treated with a regular regimen of compound glycyrrhizin tablets and glucocorticoids, respectively. Serum interleukin (IL)-18, IL-1ß, and peripheral caspase-1-positive cells were compared pre- and post-treatment among the three groups. RESULTS: The pre-treatment serum IL-1ß levels in the renal impairment and non-renal impairment groups were significantly higher than that in the control group (P<0.01). After treatment, the IL-1ß level in the non-renal impairment group was not significantly different from that in the control group (P>0.05). However, the IL-1ß level in the renal impairment group post-treatment was significantly higher than that in the other two groups (P<0.01). The positive rate of caspase-1 expression in peripheral blood before treatment in the renal impairment group and non-renal impairment group was significantly higher than that in the control group (P<0.01). After treatment, the positive rate of caspase-1 expression in the non-renal impairment group was comparable to that in the control group (P>0.05), whereas the rate in the renal impairment group was significantly higher than that in the other two groups (P<0.01). After treatment, the serum IL-1ß levels and caspase-1 positive rate in HSP patients who were responsive to treatment (as assessed by hematuria or proteinuria levels after treatment) were lower than that in patients who were unresponsive to treatment P<0.001), but not significantly different to the control group (P>0.05). CONCLUSIONS: The levels of serum IL-1ß and caspase-1 changed in response to alterations in the disease condition and treatment response in HSP patients, which suggested that pyroptosis-related inflammatory factors may have potential application value in predicting disease progression and efficacy of hormone therapy.


Subject(s)
IgA Vasculitis , Glucocorticoids , Humans , IgA Vasculitis/drug therapy , Pyroptosis
5.
Exp Cell Res ; 362(2): 504-514, 2018 01 15.
Article in English | MEDLINE | ID: mdl-29253535

ABSTRACT

The HepG2 cell line is widely used in studying liver diseases because of its immortalization, but its clinical application is limited by its low expression of the urea synthesis key enzymes and cytochromes P450 (CYPs). On the basis of our previous work, we investigated the transcriptional regulation of arginase 1 (Arg1) and ornithine transcarbamylase (OTC) in HepG2 cells. We also screened for the optimal combination of liver enrichment transcription factors (LETFs) and xenobiotic nuclear receptors that can promote the expression of key urea synthases and five major CYPs in HepG2 cells. Thus, recombinant HepG2 cells were established. Results showed that C/EBPß, not C/EBPα, could upregulate expression of Arg1 and PGC1α and HNF4α cooperatively regulate the expression of OTC. The two optimal combinations C/EBPß+HNF4α+HNF6+PXR and C/EBPß+HNF4α+HNF6+CAR were selected. Compared with the control cells, the recombinant HepG2 cells modified by the two optimal combinations exhibited enhanced ammonia metabolism and CYP enzyme activity. Moreover, the HepG2/(C/EBPß+HNF4α+HNF6+PXR) cells more strongly reduced ammonia than any other combination tested in this study. The present work indicated that optimizing the combination of transcription factors will simultaneously promote hepatocyte ammonia metabolism and drug metabolism. The recombinant HepG2 liver cell line constructed by the optimal combination provided an improved alternative means for bioartificial liver applications and drug toxicity testing.


Subject(s)
Ammonia/pharmacology , Arginase/genetics , Liver Neoplasms/metabolism , Ornithine Carbamoyltransferase/genetics , Ammonia/metabolism , Arginase/metabolism , CCAAT-Enhancer-Binding Proteins/genetics , CCAAT-Enhancer-Binding Proteins/metabolism , Gene Expression Regulation, Neoplastic , Hep G2 Cells , Hepatocyte Nuclear Factor 4/genetics , Hepatocyte Nuclear Factor 4/metabolism , Hepatocytes/drug effects , Hepatocytes/metabolism , Humans , Inactivation, Metabolic/drug effects , Inactivation, Metabolic/genetics , Liver/drug effects , Liver/metabolism , Liver Neoplasms/drug therapy , Liver Neoplasms/pathology , Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha/genetics , Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha/metabolism , Promoter Regions, Genetic/genetics
6.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-696169

ABSTRACT

Objective To explore the composition and drug resistance characteristics of pathogenic isolates from cerebrospinal fluid (CSF) cultures to facilitate empirical therapy of pediatric patients with meningitis in Guangzhou district.Methods During 2011 Jan 1st to 2015 Dec 31st,cerebrospinal fluids were collected from pediatric patients with suspected meningitis for regular culture,identification and drug susceptibility test of pathogenic isolates according to the national clinical laborato ry standard operation procedure,followed by analysis of their composition and drug resistance characteristics.Results There were 132 pathogenic isolates from CSF cultures,including Gram-positive strains (39.40%,52/132),Gram-negative strains (757.58%,6/132),fungi (3.03 %,4/132),respectively.The main isolates were Escherichia coli (E.coli) (23.48%,31/132),Streptococcus pneumonia (S.pneumonia) (22.73%,30/132),Staphylococcus aureus (S.aureus) (12.12%,16/132)and Streptococcus agalactiae (S.agalactiae) (9.85%,13/132),respectively.E.coli had no resistance to piperacillin/tazobactam,furantoin,cefepime,amikacin,tobramycin,imipenem,ertapenem and meropenem.3.22 % resistance rate to cefotetan,9.38% to ceftazidime,12.90 % to aztreonam,approximately 30 % to ampicillin/sulbactam,ofloxacin,ciprofloxacin,ceftriaxone,gentamycin,cefazolin,over 60 % to both sulfamethoxazole and ampicillin,31.25 % strains were ESBL positive.S.pneumonia had no re sis tance to ertapenem,5.88 % resistance rate to telithromycin,14.71% to chloromycetin,17.76 % to ceftriaxone,23.53 % to amoxicillin,32.30 % to meropenem,35.29 % to cefotaxime,over 70 % to tetracycline,erythromycin,penicillin,and sulfamethoxazole.S.a ureus had no resistance to rifampicin,tigecycline,quinuprin/dupletin,linezolid and vancomycin,6.25% resistance to ciprofloxacin,6.25% to gentamicin,12.50% to sulfamethoxazole,18.75% resistance to clindamycin,31.25 % to tetracycline,62.50 % to erythromycin,over 90 % to cephalosporins and nearly 100 % to penicillin,the rate of MRSA strain was 56.25 %.S.agalactiae had no resistance to penicillin,23.08 % resistance rate to ofloxacin,7.69 % to ciprofloxacin,over 60% to tetracycline,erythromycin and clindamycin.Conclusion The main pathogenic isolates from CSF cultures were E.coli,S.pneumonia,S.aureus,S.agalactiae,different species of isolates have different drug resistance characteristics.This will provide instructions for the prevention,pathogenic diagnosis and treatment of meningitis in pediatric patients.

7.
Int J Nanomedicine ; 11: 4247-59, 2016.
Article in English | MEDLINE | ID: mdl-27621619

ABSTRACT

The polyamidoamine (PAMAM) dendrimer, a type of macromolecule material, has been used in spheroidal cell culture and drug delivery in recent years. However, PAMAM is not involved in the study of hepatic cell-spheroid culture or its biological activity, particularly in detoxification function. Here, we constructed a PAMAM-dendrimer conjugate decorated by an integrin ligand: arginine-glycine-aspartic acid (RGD) peptide. Our studies demonstrate that RGD-polyethylene glycol (PEG)-PAMAM conjugates can promote singly floating hepatic cells to aggregate together in a sphere-like growth with a weak reactive oxygen species. Moreover, RGD-PEG-PAMAM conjugates can activate the AKT-MAPK pathway in hepatic cells to promote cell proliferation and improve basic function and ammonia metabolism. Together, our data support the hepatocyte sphere treated by RGD-PEG-PAMAM conjugates as a potential source of hepatic cells for a biological artificial liver system.


Subject(s)
Dendrimers/chemistry , Hepatocytes/cytology , Oligopeptides/chemistry , Polyethylene Glycols/chemistry , Spheroids, Cellular/metabolism , Cell Aggregation/drug effects , Cell Culture Techniques , Dendrimers/metabolism , Hepatocytes/drug effects , Hepatocytes/metabolism , Humans , Oligopeptides/metabolism , Polyethylene Glycols/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Reactive Oxygen Species/metabolism
8.
Plant Cell ; 27(5): 1445-60, 2015 May.
Article in English | MEDLINE | ID: mdl-25966761

ABSTRACT

Drought is a major threat to plant growth and crop productivity. Calcium-dependent protein kinases (CDPKs, CPKs) are believed to play important roles in plant responses to drought stress. Here, we report that Arabidopsis thaliana CPK8 functions in abscisic acid (ABA)- and Ca(2+)-mediated plant responses to drought stress. The cpk8 mutant was more sensitive to drought stress than wild-type plants, while the transgenic plants overexpressing CPK8 showed enhanced tolerance to drought stress compared with wild-type plants. ABA-, H2O2-, and Ca(2+)-induced stomatal closing were impaired in cpk8 mutants. Arabidopsis CATALASE3 (CAT3) was identified as a CPK8-interacting protein, confirmed by yeast two-hybrid, coimmunoprecipitation, and bimolecular fluorescence complementation assays. CPK8 can phosphorylate CAT3 at Ser-261 and regulate its activity. Both cpk8 and cat3 plants showed lower catalase activity and higher accumulation of H2O2 compared with wild-type plants. The cat3 mutant displayed a similar drought stress-sensitive phenotype as cpk8 mutant. Moreover, ABA and Ca(2+) inhibition of inward K(+) currents were diminished in guard cells of cpk8 and cat3 mutants. Together, these results demonstrated that CPK8 functions in ABA-mediated stomatal regulation in responses to drought stress through regulation of CAT3 activity.


Subject(s)
Abscisic Acid/metabolism , Arabidopsis Proteins/metabolism , Arabidopsis/enzymology , Calcium/metabolism , Cyclin-Dependent Kinase 8/metabolism , Hydrogen Peroxide/metabolism , Plant Growth Regulators/metabolism , Signal Transduction , Arabidopsis/genetics , Arabidopsis/physiology , Arabidopsis Proteins/genetics , Catalase/genetics , Catalase/metabolism , Cyclin-Dependent Kinase 8/genetics , Droughts , Gene Expression Regulation, Plant , Homeostasis , Plant Stomata/enzymology , Plant Stomata/genetics , Plant Stomata/physiology , Plants, Genetically Modified , Protein Kinases/genetics , Protein Kinases/metabolism , Stress, Physiological
9.
Mol Plant ; 6(5): 1487-502, 2013 Sep.
Article in English | MEDLINE | ID: mdl-23404561

ABSTRACT

The Arabidopsis Di19 (Drought-induced) gene family encodes seven Cys2/His2-type zinc-finger proteins, most with unknown functions. Here, we report that Di19 functioned as a transcriptional regulator and was involved in Arabidopsis responses to drought stress through up-regulation of pathogenesis-related PR1, PR2, and PR5 gene expressions. The Di19 T-DNA insertion mutant di19 was much more sensitive to drought stress, whereas the Di19-overexpressing lines were much more tolerant to drought stress compared with wild-type plants. Di19 exhibited transactivation activity in our yeast assay, and its transactivation activity was further confirmed in vivo. DNA-binding analysis revealed that Di19 could bind to the TACA(A/G)T element and chromatin immunoprecipitation (ChIP) assays demonstrated that Di19 could bind to the TACA(A/G)T element within the PR1, PR2, and PR5 promoters. qRT-PCR results showed that Di19 promoted the expressions of PR1, PR2, and PR5, and these heightened expressions were enhanced by CPK11, which interacted with Di19 in the nucleus. Similarly to the Di19-overexpressing line, PR1-, PR2-, and PR5-overexpressing lines also showed the drought-tolerant phenotype. The pre-treatment with salicylic acid analogs INA can enhance plants' drought tolerance. Taken together, these data demonstrate that Di19, a new type of transcription factor, directly up-regulates the expressions of PR1, PR2, and PR5 in response to drought stress, and its transactivation activity is enhanced by CPK11.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/genetics , Arabidopsis/physiology , Carrier Proteins/metabolism , Droughts , Stress, Physiological/genetics , Adaptation, Physiological/genetics , Arabidopsis Proteins/genetics , Base Sequence , Gene Expression Regulation, Plant , Genes, Plant/genetics , Glucan Endo-1,3-beta-D-Glucosidase/genetics , Glucan Endo-1,3-beta-D-Glucosidase/metabolism , Models, Biological , Molecular Sequence Data , Mutation/genetics , Phenotype , Promoter Regions, Genetic/genetics , Protein Binding/genetics , Protein Kinases/metabolism , Subcellular Fractions/drug effects , Subcellular Fractions/metabolism , Transcription Factors/metabolism , Transcriptional Activation/genetics , Up-Regulation/genetics
10.
Dalton Trans ; 41(34): 10422-9, 2012 Sep 14.
Article in English | MEDLINE | ID: mdl-22814858

ABSTRACT

A microporous Pb(II) metal-organic framework (MOF) [PbL(2)]·2DMF·6H(2)O (1) has been assembled from a N-oxide and amide doubly functionalized ligand HL (= N-(4-carboxyphenyl)isonicotinamide 1-oxide). Complex 1 features a three-dimensional (3D) framework possessing one-dimensional (1D) rhombic channels with dimensions of 13 × 13 Å(2). The 3D framework is built up from 1D PbO(2) chains that link ligands in parallel fashion to construct single-wall channels. When recrystallizing 1 in a DMSO-DMF mixture (3 : 5 v/v), a new coordination polymer, [PbL(2)]·DMF·2H(2)O (2), was obtained. Complex 2 is also a 3D framework containing 1D rectangular channels, but the channel dimensions become reduced in size to 13 × 8 Å(2) due to reorganization of the Pb(ii) coordination environment. The PbO(2) chains in 2 are reformed to link ligands in a double-wall fashion, significantly reducing the channel size. Even though, the guest exchange study indicates that the DMF molecules in 2 could be replaced with benzene molecules when immersing in benzene solvent, showing single-crystal-to-single-crystal (SC-SC) guest exchange in the solid state and leading to a daughter crystal [PbL(2)]·0.5C(6)H(6)·2H(2)O (2'). Desolvated 1 and 2 display preferential adsorption behaviors of water vapour over CO(2) due to the hydrophilic nature of the channels and the strong host-guest interactions. Catalytic tests indicate that desolvated 1 and 2 have size-selective catalytic activity towards the Knoevenagel condensation reaction.

11.
Zhong Yao Cai ; 32(8): 1256-9, 2009 Aug.
Article in Chinese | MEDLINE | ID: mdl-19960951

ABSTRACT

OBJECTIVE: To study the inhibiting effect of Vaccaria segetalis extracts on neovascularization. METHODS: The effect of Vaccaria segetalis extracts on the proliferation, migration in vitro and tube formation on Matrigel of endothelial cell (HMEC) in vivo were examined by MTT assay and Matrigel plug assay. RESULTS: The proliferation and migration of HMEC were inhibited significantly by Vaccaria segetalis extracts in a dose-dependent manner (IC50 = 50 microg/mL). It also inhibited angiogenesis in Matrigel plug mouse model. CONCLUSION: Vaccaria segetalis extracts can inhibit angiogenensis obviously, and it could be developed as an effective antiangiogenic drug.


Subject(s)
Angiogenesis Inhibitors/pharmacology , Endothelial Cells/drug effects , Neovascularization, Pathologic , Plant Extracts/pharmacology , Vaccaria/chemistry , Angiogenesis Inhibitors/administration & dosage , Animals , Cell Movement/drug effects , Cell Proliferation/drug effects , Cells, Cultured , Collagen , Dose-Response Relationship, Drug , Drug Combinations , Female , Fibroblast Growth Factor 2/administration & dosage , Fibroblast Growth Factor 2/pharmacology , Flow Cytometry , Humans , Laminin , Mice , Mice, Nude , Plant Extracts/administration & dosage , Proteoglycans , Random Allocation
12.
J Biosci ; 34(2): 213-20, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19550037

ABSTRACT

ADAM15 plays an important role in tumour development by interacting with integrins. In this study, we investigated the target peptides of the ADAM15 disintegrin domain. First, we successfully produced the recombinant human ADAM15 disintegrin domain (RADD) that could inhibit melanoma cell adhesion by using Escherichia coli. Second, four specific binding peptides (peptides A, B, C, and D) were selected using a phage display 12-mer peptide library. The screening protocol involved 4 rounds of positive panning on RADD and 2 rounds of subtractive selection with streptavidin. By using the BLAST software and a relevant protein database, integrin alpha v beta 3 was found to be homologous to peptide A. Synthetic peptide A had a highly inhibitory effect on RADD-integrin alpha v beta 3 binding. The results demonstrate the potential application of short peptides for disrupting high-affinity ADAM-integrin interactions.


Subject(s)
ADAM Proteins/physiology , Membrane Proteins/physiology , ADAM Proteins/chemistry , Cell Adhesion , Cell Line, Tumor , Dose-Response Relationship, Drug , Enzyme-Linked Immunosorbent Assay , Escherichia coli/metabolism , Humans , Integrins/metabolism , Melanoma/metabolism , Membrane Proteins/chemistry , Models, Biological , Peptide Library , Peptides/chemistry , Protein Binding , Protein Structure, Tertiary , Proteomics/methods
13.
Phytomedicine ; 16(8): 703-11, 2009 Aug.
Article in English | MEDLINE | ID: mdl-19423313

ABSTRACT

PURPOSE: The purpose of this study was to investigate the anti-angiogenic properties of julibroside J(8), a triterpenoid saponin isolated from Albizia julibrissin. METHODS: In the presence of juliborside J(8,) the growth of human microvascular endothelial cells (HMEC-1), four human tumor cell lines, and a normal cell line (MRC-5) was evaluated by MTT assay. The in vivo anti-angiogenic effect of julibroside J(8) was evaluated on a chorioallantoic membrane (CAM) and in transplanted colon carcinoma cells in a nude mice neovascularisation model. RESULTS: Treatment with 0.5-4 microg/ml julibroside J(8) resulted in dose-dependent inhibition of growth, migration, and tube formation in HMEC-1 cells; julibroside J(8) also inhibited the formation of microvessels on CAM at a concentration of 10-50 microg/egg and reduced vessel density within tumor at a concentration of 0.5-3mg/kg. CONCLUSIONS: Julibroside J(8) may be a potent anti-angiogenetic and cytotoxic drug; further investigation is warranted.


Subject(s)
Albizzia/chemistry , Angiogenesis Inhibitors/pharmacology , Antineoplastic Agents, Phytogenic/pharmacology , Plant Extracts/pharmacology , Polysaccharides/pharmacology , Saponins/pharmacology , Angiogenesis Inhibitors/chemistry , Angiogenesis Inhibitors/therapeutic use , Animals , Antineoplastic Agents, Phytogenic/chemistry , Antineoplastic Agents, Phytogenic/therapeutic use , Cell Line , Cell Line, Tumor , Chick Embryo , Chorioallantoic Membrane , Colonic Neoplasms , Dose-Response Relationship, Drug , Endothelial Cells , Humans , Mice , Microvessels/drug effects , Phytotherapy , Plant Bark , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Plant Extracts/therapeutic use , Plant Stems , Plants, Medicinal/chemistry , Polysaccharides/chemistry , Polysaccharides/isolation & purification , Polysaccharides/therapeutic use , Saponins/therapeutic use
14.
Plant Signal Behav ; 4(12): 1193-5, 2009 Dec.
Article in English | MEDLINE | ID: mdl-20514245

ABSTRACT

Pollen germination (PG) and pollen tube growth (PTG) play crucial roles in sexual reproduction of flowering plants by sending sperm cells to the ovule. These two processes are regarded as ideal model system for the study of cell signaling and cell polarized growth. It has been considered for a long time that ion transports across the pollen tube membranes are essential for pollen tube navigation and growth. Previous transcriptome analyses for Arabidopsis have shown that the transcripts related to cellular transport are correspondingly overrepresented during the process of pollen tube growth. Here, we showed that 459 transporter genes expressed during PG and PTG in Arabidopsis. In addition, the gene expression profiles of ion (including Ca(2+), H(+), K(+), Cl(-)) channels and transporters were further analyzed. This analysis provides novel information for the potential candidate genes involving in ion fluxes across the pollen tube membranes and in regulation of pollen tube tip growth.

15.
Plant Physiol ; 148(3): 1201-11, 2008 Nov.
Article in English | MEDLINE | ID: mdl-18775970

ABSTRACT

Pollen germination, along with pollen tube growth, is an essential process for the reproduction of flowering plants. The germinating pollen with tip-growth characteristics provides an ideal model system for the study of cell growth and morphogenesis. As an essential step toward a detailed understanding of this important process, the objective of this study was to comprehensively analyze the transcriptome changes during pollen germination and pollen tube growth. Using Affymetrix Arabidopsis (Arabidopsis thaliana) ATH1 Genome Arrays, this study is, to our knowledge, the first to show the changes in the transcriptome from desiccated mature pollen grains to hydrated pollen grains and then to pollen tubes of Arabidopsis. The number of expressed genes, either for total expressed genes or for specifically expressed genes, increased significantly from desiccated mature pollen to hydrated pollen and again to growing pollen tubes, which is consistent with the finding that pollen germination and tube growth were significantly inhibited in vitro by a transcriptional inhibitor. The results of Gene Ontology analyses showed that expression of genes related to cell rescue, transcription, signal transduction, and cellular transport was significantly changed, especially for up-regulation, during pollen germination and tube growth. In particular, genes of the calmodulin/calmodulin-like protein, cation/hydrogen exchanger, and heat shock protein families showed the most significant changes during pollen germination and tube growth. These results demonstrate that the overall transcription of genes, both in the number of expressed genes and in the levels of transcription, was increased. Furthermore, the appearance of many novel transcripts during pollen germination as well as tube growth indicates that these newly expressed genes may function in this complex process.


Subject(s)
Arabidopsis/genetics , Gene Expression , Germination , Pollen/metabolism , RNA, Messenger/genetics , Arabidopsis/growth & development , Dactinomycin/pharmacology , Genes, Plant , Polymerase Chain Reaction , Transcription, Genetic/drug effects
16.
Protein Expr Purif ; 61(1): 45-9, 2008 Sep.
Article in English | MEDLINE | ID: mdl-18556214

ABSTRACT

Glucagon-like peptide-1 (GLP-1) is a 30-residue peptide hormone secreted by intestinal L-cells in response to nutrient ingestion. In the present study, overlapping PCR technology was employed to construct two GLP-1 mutants (GLP-1(A2G))2 and human albumin (HSA) genes in vitro without linker. The spliced gene, (GLP-1(A2G))2-HSA, was over expressed under the control of promoter AOX1 and Mat alpha signal peptide in Pichia pastoris. SDS-PAGE and Western blotting were applied to assay the recombinant fusion protein in the culture broth. The results demonstrated that the recombinant (GLP-1(A2G))2-HSA concentration in the broth could reach a level of 245.0 mg/L and the expressed fusion protein was capable of cross-reacting with anti-human GLP-1 and anti-human albumin antibody. The recombinant (GLP-1(A2G))2-HSA protein was purified by ultrafiltration, columns of Q-sepharose fast flow and Superdex 75 size-exclusion. The recombinant (GLP-1(A2G))2-HSA protein obtained could lower in vivo glucose concentration in blood and stimulate in vitro islet cell proliferation. In mouse model, the fusion protein was detectable in plasma even 308 h after a single subcutaneous dose of 1.25 mg/kg. The result showed that the terminal biological half-time of the protein was about 54.2 h which is 650-fold longer than that of GLP-1. The pharmacokinetic analysis of the protein suggests its promising application in clinical medicine.


Subject(s)
Glucagon-Like Peptide 1/genetics , Mutation , Pichia/genetics , Recombinant Fusion Proteins/genetics , Serum Albumin/genetics , Base Sequence , Blotting, Western , Chromatography, High Pressure Liquid , DNA Primers , Electrophoresis, Polyacrylamide Gel , Humans , Recombinant Fusion Proteins/chemistry , Recombinant Fusion Proteins/isolation & purification
17.
Appl Radiat Isot ; 66(12): 1881-5, 2008 Dec.
Article in English | MEDLINE | ID: mdl-18571931

ABSTRACT

2Beta-carbomethoxy-3beta-(4-chlorophenyl)-8-(2-[(18)F]fluoroethyl)nortropane ((18)F-FECNT) is a potential dopamine transporter imaging agent. In this article, a new mesylate precursor was prepared and a one-step automated synthesis of (18)F-FECNT was developed. The mesylate precursor (4) was synthesized from 2beta-carbomethoxy-3beta-(4-chlorophenyl)tropane (1) by N-demethylation, hydroxyethylation followed by mesylation at a total yield of 47%. [(18)F]fluorination was performed by heating 4mg mesylate precursor and K[(18)F] in 1 ml acetonitrile at 90 degrees C for 20 min. The crude (18)F-FECNT was obtained with a radiolabeling yield of 48%. After purification by preparative high performance liquid chromatography (HPLC), the final (18)F-FECNT product was obtained with a radiochemical purity of 98.4% and a decay corrected radiochemical yield of 33+/-9% (and the uncorrected radiochemical yield was 19+/-5%, n=5). The duration of the total procedure was 80-90 min.


Subject(s)
Isotope Labeling/instrumentation , Mesylates/chemistry , Nortropanes/chemistry , Nortropanes/isolation & purification , Radiopharmaceuticals/chemical synthesis , Radiopharmaceuticals/isolation & purification , Robotics/instrumentation
18.
Trans R Soc Trop Med Hyg ; 101(11): 1157-8, 2007 Nov.
Article in English | MEDLINE | ID: mdl-17606284

ABSTRACT

The level of Toxoplasma gondii infection among the general population of the Democratic Republic of Sao Tome and Principe is unclear. The T. gondii infection status of inhabitants who visited National Central Hospital on Sao Tome Island was assessed by a latex agglutination test. The overall seroprevalence was 74.5% (120/161). No significant gender difference in seroprevalence was found between males and females. The older age group (> or =45 years) had significantly higher seroprevalence (80.0%, 28/35) than the younger age group (<15 years) (20.0%, 3/15) (chi(2)=16.04, P<0.001).


Subject(s)
Antibodies, Protozoan/blood , Toxoplasmosis/diagnosis , Adolescent , Adult , Age Distribution , Animals , Atlantic Islands/epidemiology , Female , Humans , Latex Fixation Tests/methods , Male , Middle Aged , Pregnancy , Pregnancy Complications, Parasitic/diagnosis , Pregnancy Complications, Parasitic/epidemiology , Prevalence , Toxoplasmosis/epidemiology
19.
Trans R Soc Trop Med Hyg ; 101(2): 134-9, 2007 Feb.
Article in English | MEDLINE | ID: mdl-17113117

ABSTRACT

The seroprevalence of Toxoplasma gondii infection among pregnant women in the Democratic Republic of Sao Tome and Principe (DRSTP) from November 2003 to March 2004 was determined by detection of serum anti-T. gondii antibodies. A short questionnaire interview for pregnant women was performed to investigate risk factors associated with T. gondii infection, including consumption of raw meat or unwashed vegetables, drinking unboiled water and keeping pets (cats and dogs). The overall seroprevalence of T. gondii infection was high (75.2%; 375/499). The older age group of > or =35 years had a significantly higher seroprevalence (85.7%; 54/63) than that of the younger age group of 15-25 years (70.4%; 178/253) (odds ratio 2.5, 95% CI 1.2-5.4; P=0.01). No significant difference in the seroprevalence of T. gondii infection was found between the pregnant women with and without exposure to the risk factors studied. However, among pregnant women with high antibody titers of > or =1:1024, it seemed likely that continual contact with pets and consumption of oocyst-contaminated water or raw unwashed vegetables rather than tissue cysts in meat was the primary route of infection. The incidence of congenital toxoplasmosis in unborn babies should be intensively monitored in the DRSTP.


Subject(s)
Pregnancy Complications, Parasitic/epidemiology , Toxoplasmosis, Congenital/epidemiology , Toxoplasmosis/epidemiology , Adolescent , Adult , Animals , Animals, Domestic , Antibodies, Protozoan , Atlantic Islands/epidemiology , Child , Female , Humans , Latex Fixation Tests , Meat Products , Pregnancy , Prevalence , Regression Analysis , Risk Factors , Toxoplasma/isolation & purification , Water/parasitology
20.
Yao Xue Xue Bao ; 41(9): 814-8, 2006 Sep.
Article in Chinese | MEDLINE | ID: mdl-17111825

ABSTRACT

AIM: To study the effect of recombinant hirudin (rH) on tPA-induced fibrinolysis and the possible mechanism of its action. METHODS: The effect of rH on thrombin-fibrin complex (Th-Fn) was detected by 99mTc labeled rH. In the in vitro clot lysis, tPA as plasminogen activator, and recalcified plasma as plasminogen resource were used to study the influence of rH on fibrinolysis by detecting TAFIa, D-Dimer and FXIII. RESULTS: In a canine model of femoral artery thrombosis, a clear radioactivity strip was imaged in 30 - 60 min on a part image, and the femoral vein thrombosis developed at 30 min. rH efficiently inhibited clot regeneration. Addition of TM could inhibit clot lysis obviously, and CPI could shorten the delay of clot lysis which due to TAFIa. There was a dose-dependent relationship with TM concentration and TAFI activation. FXIII activation was inhibited by low concentration of rH ( < or = 0.2 u x mL(-1)), and the level of fibrinolysis product, D-Dimer, increased. CONCLUSION: rH could inhibit the thrombin binding to fibrin. rH inhibited the activation of TAFI and FXIII by combining with thrombin which resulted in enhancement of thrombolysis.


Subject(s)
Carboxypeptidase B2/metabolism , Fibrinolysis/drug effects , Hirudins/pharmacology , Thrombosis/metabolism , Animals , Blood Coagulation/drug effects , Carboxypeptidases/antagonists & inhibitors , Dogs , Factor XIII/metabolism , Femoral Artery , Femoral Vein , Fibrinolytic Agents/pharmacology , Hirudins/genetics , Male , Plant Proteins/pharmacology , Protease Inhibitors , Recombinant Proteins/pharmacology , Thrombomodulin/metabolism , Venous Thrombosis/metabolism
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