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1.
Int J Ophthalmol ; 17(5): 869-876, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38766331

RESUMO

AIM: To investigate the difference in risk factors between non-arteritic anterior ischaemic optic neuropathy (NAION) and central retinal artery occlusion (CRAO) and develop a predictive diagnostic nomogram. METHODS: The study included 37 patients with monocular NAION, 20 with monocular CRAO, and 24 with hypertension. Gender, age, and systemic diseases were recorded. Blood routine, lipids, hemorheology, carotid and brachial artery doppler ultrasound, and echocardiography were collected. The optic disc area, cup area, and cup-to-disc ratio (C/D) of the unaffected eye in the NAION and CRAO group and the right eye in the hypertension group were measured. RESULTS: The carotid artery intimal medial thickness (C-IMT) of the affected side of the CRAO group was thicker (P=0.039) and its flow-mediated dilation (FMD) was lower (P=0.049) than the NAION group. Compared with hypertension patients, NAION patients had higher whole blood reduced viscosity low-shear (WBRV-L) and erythrocyte aggregation index (EAI; P=0.045, 0.037), and CRAO patients had higher index of rigidity of erythrocyte (IR) and erythrocyte deformation index (EDI; P=0.004, 0.001). The optic cup and the C/D of the NAION group were smaller than the other two groups (P<0.0001). The diagnostic prediction model showed high diagnostic specificity (83.7%) and sensitivity (85.6%), which was highly related to hypertension, the C-IMT of the affected side, FMD, platelet (PLT), EAI, and C/D. CONCLUSION: CRAO patients show thicker C-IMT and worse endothelial function than NAION. NAION and CRAO may be related to abnormal hemorheology. A small cup and small C/D may be involved in NAION. The diagnostic nomogram can be used to preliminarily identify NAION and CRAO.

2.
Phytomedicine ; 114: 154774, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36996530

RESUMO

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is currently the third leading cause of death globally. Oxidative stress affects various molecular mechanisms and is the main driving factor of COPD. Ally isothiocyanate (AITC) is an effective component of Semen Sinapis Albae, which has favorable effects for the treatment of COPD, but its mechanism has not been fully elucidated. PURPOSE: This study aimed to elucidate the antioxidant effect of AITC on COPD and its molecular mechanism, and preliminarily determine the role of AhR in the progression of COPD. STUDY DESIGN: The COPD rat model was established by smoking combined with intratracheal instillation of lipopolysaccharide. Different doses of AITC, positive control drug acetylcysteine, AhR inhibitor alpha-naphthoflavone, and agonist beta-naphthoflavone were administered by gavage. Human bronchial epithelial cells induced by cigarette smoke extract (CSE) were used in an in vitro model to explore the molecular mechanisms of AITC. METHODS: The effects of AITC on lung function and oxidative stress in rats were evaluated in vivo using the respiratory function test, white blood cell count, enzyme-linked immunosorbent assay, and histological staining. The changes in protein expression in the lung tissue were detected by immunohistochemistry and Western blotting. RT-PCR, western blotting, and immunofluorescence were used to explore the molecular mechanisms of AITC. Enzyme-linked immunosorbent assay, reactive oxygen species probing, and flow cytometry were used to determine the antioxidant effect of AITC. RESULTS: AITC can improve the lung function of rats with COPD, restore lung tissue structure, improve oxidative stress, reduce inflammation, and inhibit lung cell apoptosis. AITC reversed the upregulation of AhR and CYP1A1 and the down-regulation of Nrf2 and NQO1 in the lung tissues of rats with COPD. CSE stimulation can increase the expressions of AhR and CYP1A1 and decrease the expressions of Nrf2 and NQO1 in 16HBE cells, leading to severe oxidative stress and inflammatory response and, ultimately, apoptosis. AITC inhibited AhR and CYP1A1 expressions, induced Nrf2 and NQO1 expressions, promoted Nrf2 nuclear translocation, and improved CSE-induced toxicological effects. CONCLUSION: AITC may improve lung oxidative stress by inhibiting the AhR / CYP1A1 and activating the Nrf2 / NQO1 pathways, thereby delaying the pathological progression of COPD.


Assuntos
Fator 2 Relacionado a NF-E2 , Doença Pulmonar Obstrutiva Crônica , Ratos , Humanos , Animais , Fator 2 Relacionado a NF-E2/metabolismo , Citocromo P-450 CYP1A1/metabolismo , Antioxidantes/farmacologia , Transdução de Sinais , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Isotiocianatos/farmacologia , Estresse Oxidativo , NAD(P)H Desidrogenase (Quinona)/metabolismo
3.
Artif Cells Nanomed Biotechnol ; 49(1): 699-708, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34882059

RESUMO

Spinal cord injury (SCI) is a common pathology often resulting in permanent loss of sensory, motor, and autonomic function. Numerous studies in which stem cells have been transplanted in biomaterial scaffolds into animals have demonstrated their considerable potential for recovery from SCI. In the present study, a three-dimensional porous silk fibroin (SF) scaffold with a mean pore size of approximately 383 µm and nanofibrous structure was fabricated, the silk scaffold enabling the enhanced attachment and proliferation of bone marrow stromal cells (BMSCs). Investigation of its therapeutic potential was conducted by implantation of the nanofibrous SF scaffold seeded with BMSCs into a transected spinal cord model. Recovery of the damaged spinal cord was significantly improved after 2 months, compared with a non-nanofibrous scaffold, in combination with decreased glial fibrillary acidic protein (GFAP) expression and improved axonal regeneration at the site of injury. Furthermore, elevated Basso-Beattie-Bresnahan (BBB) scores indicated greatly improved hindlimb movement. Together, these results demonstrate that transplantation of neural scaffolds consisting of nanofibrous SF and BMSCs is an attractive strategy for the promotion of functional recovery following SCI.


Assuntos
Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais , Nanofibras , Traumatismos da Medula Espinal , Regeneração da Medula Espinal , Animais , Células da Medula Óssea , Transplante de Células-Tronco Mesenquimais/métodos , Nanofibras/química , Ratos , Ratos Sprague-Dawley , Recuperação de Função Fisiológica , Seda/química , Medula Espinal , Traumatismos da Medula Espinal/terapia , Alicerces Teciduais/química
4.
Int Immunopharmacol ; 101(Pt B): 108373, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34802946

RESUMO

BACKGROUND AND AIMS: Allyl isothiocyanate(AITC) has been shown to play an important role in the improved symptoms of chronic obstructive pulmonary disease(COPD) and the inhibition of inflammation, but the role in COPD lipid metabolism disorder and the molecular mechanism remains unclear. We aimed to explore whether and how AITC affects COPD by regulating lipid metabolism and inflammatory response. METHODS: The COPD rat model was established by cigarette smoke exposure. Cigarette smoke extract stimulated 16HBE cells to induce a cell model. The effect of AITC treatment was detected by lung function test, H&E staining, Oil red O staining, immunohistochemistry, ELISA, CCK-8, HPLC, fluorescence efflux test, siRNA, RT-PCR, and Western blotting. Biological analysis was performed to analyze the results. Graphpad Prism 8.0 software was used for statistical analysis. RESULTS: AITC can improve lung function and pathological injury in COPD rats. The levels of IL-1 ß and TNF- α in the AITC treatment group were significantly lower than those in the model group(P < 0.05), and the lipid metabolism was also improved (P < 0.05). AITC reverses CSE-induced down-regulation of LXR α, ABCA1, and ABCG1 expression and function in a time-and concentration-dependent manner (P < 0.05). AITC regulates the cholesterol metabolism disorder induced by CSE in NR8383 cells and attenuates macrophage inflammation (P < 0.05). In addition, after silencing LXR α with siRNA, the effect of AITC was also inhibited. CONCLUSION: These results suggest that AITC improves COPD by promoting RCT process and reducing inflammatory response via activating LXR pathways.


Assuntos
Transportador 1 de Cassete de Ligação de ATP/metabolismo , Membro 1 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Isotiocianatos/farmacologia , Doença Pulmonar Obstrutiva Crônica/induzido quimicamente , Fumaça/efeitos adversos , Transportador 1 de Cassete de Ligação de ATP/genética , Membro 1 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/genética , Animais , Linhagem Celular , Conservantes de Alimentos/farmacologia , Receptores X do Fígado/agonistas , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Masculino , Doença Pulmonar Obstrutiva Crônica/metabolismo , Ratos , Ratos Sprague-Dawley , Regulação para Cima
5.
J Pediatr Endocrinol Metab ; 34(9): 1147-1156, 2021 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-34187135

RESUMO

OBJECTIVES: To report two novel mutations in the BCKDHB gene with Maple syrup urine disease (MSUD) and compare their data with 52 cases of MSUD reported in the available Chinese literature. METHODS: Clinical data of a case of a newborn with MSUD was retrospectively studied. Literatures on MSUD in the local medical journals from January 1990 till December 2019 in China were reviewed. RESULTS: Two novel BCKDHB mutations c.90_91insCTGGCGCGGGG (p.Phe35TrpfsTer41) and c.80_90del (p.Ala32PhefsTer48) were identified. We found a total of 52 cases of MSUD reports so far. A total of 49 cases had the symptom of poor feeding (94.2%), 50 cases showed poor responses to stimulation (96.2%), 21 cases had odor of maple syrup (40.3%), 29 cases had seizures (55.7%), and 13 cases had respiratory failure (25.0%). The average of the blood ammonia was 127.2 ± 75.0 µmol/L. A total of 18 cases reported the gene testing, among of them 9 cases of BCKDHA mutations, 6 cases of BCKDHB mutations, and 2 cases of DBT mutations. A total of 13 cases (25%) were treated with mechanical ventilation, 50 cases (96.2%) with protein-restricted diet and l-carnitine, 29 cases with thiamine, and only 2 cases were treated with blood purification. Finally, 19 patients (36.5%) were died, 21 cases (40.4%) were improved after treatments. CONCLUSIONS: The clinical phenotype of neonatal MSUD in China belongs to the classical type currently. Suspected patients should have blood or urine branched-chain amino acid levels tested and brain MRI as early as possible to enable early diagnosis, thus improvement in prognosis.


Assuntos
3-Metil-2-Oxobutanoato Desidrogenase (Lipoamida)/genética , Doença da Urina de Xarope de Bordo/patologia , Mutação , Fenótipo , China/epidemiologia , Feminino , Seguimentos , Testes Genéticos , Humanos , Recém-Nascido , Masculino , Doença da Urina de Xarope de Bordo/epidemiologia , Doença da Urina de Xarope de Bordo/genética , Doença da Urina de Xarope de Bordo/terapia , Prognóstico , Estudos Retrospectivos
6.
Cancer Biol Ther ; 20(6): 866-876, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30786815

RESUMO

Most of gastric carcinoma (GC) is attributed to infection by Helicobacter pylori (H. pylori) but there is increasing evidence that the positive H. pylori status correlates with better prognosis in GC. The H. pylori-induced cellular immune response may suppress cancer and in this work, recombinant pcDNA3 plasmids encoding various fragments of H. pylori virulence genes of cagA, vacA and babA are constructed and combined into groups to immunize BALB/c mice. The activated splenic CD3+ T cells are purified and the anticancer effects are investigated in vitro and in vivo. The H. pylori DNA vaccines induce a shift in the response from Th1 to Th2 that mimicks the immune status in patients of GC with chronic H. pylori infection. The stimulated CD3+ T cells inhibit the growth of human GC cells in vitro and adoptive transfusions of the CD3+ T cells suppress the growth of GC xenograft in vivo. The effects may be caused by the larger ratios of infiltrated CD8+/CD4+ T cells, reduced infiltration of regulatory FOXP3+ T cells, and enhanced apoptosis induced by upregulation of Caspase-9/Caspase-3 and downregulation of Survivin. Our results reveal the potential immunotherapeutic value of H. pylori vaccine-activated CD3+ T cells in those with advanced GC.


Assuntos
Vacinas Bacterianas/imunologia , Helicobacter pylori/imunologia , Ativação Linfocitária/imunologia , Subpopulações de Linfócitos T/imunologia , Vacinas de DNA/imunologia , Animais , Apoptose/genética , Apoptose/imunologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Vacinas Bacterianas/administração & dosagem , Infecções por Helicobacter/imunologia , Infecções por Helicobacter/prevenção & controle , Helicobacter pylori/genética , Interações Hospedeiro-Patógeno/imunologia , Humanos , Imunoterapia/métodos , Linfócitos do Interstício Tumoral/imunologia , Linfócitos do Interstício Tumoral/metabolismo , Camundongos , Neoplasias Gástricas/etiologia , Neoplasias Gástricas/terapia , Subpopulações de Linfócitos T/metabolismo , Vacinas de DNA/administração & dosagem
7.
Zhongguo Zhong Yao Za Zhi ; 44(24): 5465-5472, 2019 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-32237396

RESUMO

The aim of this paper was to screen the active targets of Schizonepetae Herba and Saposhnikoviae Radix in the treatment of ulcerative colitis by means of network pharmacology,and to investigate their mechanism of action. The effective components of Schizonepetae Herba and Saposhnikoviae Radix were screened out by traditional Chinese medicine systematic pharmacological( TCMSP)database,with oral bioavilability( OB) ≥30% and drug-like( DL) ≥18% selected as the thresholds. Target PPI network was built between the main components and their corresponding targets. One hundred and eighty-two human genes corresponding to the medicine target sites were obtained from Uniprot database; 3 874 genes corresponding to ulcerative colitis were obtained from Genecard database.A total of 115 intersection genes were screened from disease genes and medicine genes,and the PPI interaction analysis was conducted by using String tool. Disease-target PPI network was drawn by using Cytoscape software,and component-target-disease network was constructed. One hundred and eight nodes and 1 882 connections were found,and then Cytoscape software was used to merge the networks and filter the core network for gene GO function analysis and KEGG pathway enrichment analysis. The mechanism of Schizonepetae Herba and Saposhnikoviae Radix was then verified by animal experiment. Gene GO functional analysis suggested that biological process,molecular functions and cell components were involved,and it was found that ulcerative colitis might be related to transcription factor activity,and cytokine receptor binding,etc. Gene KEGG pathway enrichment analysis showed that the mechanism of ulcerative colitis might be associated with TNF and Toll-like receptors( TLRs) signaling pathway-mediated cytoinflammatory factors interleukin-1( IL-1) and interleukin-6( IL6). The possible mechanism of the effective components of Schizonepetae Herba and Saposhnikoviae Radix in treating ulcerative colitis might be related to intervening the cytokine receptor binding of TNF and TLRs signaling pathways,reducing the transcription of nuclear factor-kappaB( NF-κB),and inhibiting the secretion of intestinal inflammatory factors IL-1 and IL-6.


Assuntos
Colite Ulcerativa/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Mapeamento de Interação de Proteínas , Animais , Apiaceae/química , Bases de Dados Genéticas , Humanos , Interleucinas/metabolismo , Lamiaceae/química , Medicina Tradicional Chinesa , Fitoterapia , Raízes de Plantas/química , Transdução de Sinais , Software , Receptores Toll-Like/metabolismo
8.
Neuroscience ; 390: 151-159, 2018 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-30170158

RESUMO

Localization of apelin and its receptor APJ in limbic structures such as the hippocampus suggests potential involvement of apelin/APJ signaling in stress-related emotional responses. We have recently reported that apelin-13 exerts antidepressant-like actions in acute stressed rats, and that the hippocampus is a critical brain region mediating its actions. However, the neural mechanism underling antidepressant-like actions of apelin-13 is still largely unknown. The aim of the present study is to determine whether apelin-13 ameliorates chronic water-immersion restraint stress (CWIRS)-induced depression-like phenotypes and its neural mechanism in rats. Here, we report that CWIRS exposure leaded to upregulation of apelin/APJ signaling in the hippocampus. Apelin-13 ameliorated CWIRS-induced depression-like phenotypes including hedonic-like deficit and behavioral despairs. Moreover, apelin-13 ameliorated hypothalamic-pituitary-adrenal (HPA) axis hyperactivity, and hippocampal BDNF expression deficit and glucocorticoid receptor (GR) nucleus translocation hypoactivity in chronic stressed rats. Finally, apelin-13-mediated effects were blocked by the selective TrkB receptor antagonist ANA-12. These results suggest that apelin-13 upregulates BDNF against chronic stress-induced depression-like phenotypes by ameliorating HPA axis and hippocampal GR dysfunctions.


Assuntos
Apelina/metabolismo , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Depressão/metabolismo , Hipocampo/metabolismo , Hipotálamo/metabolismo , Sistema Hipófise-Suprarrenal/metabolismo , Receptores de Glucocorticoides/metabolismo , Estresse Psicológico/metabolismo , Animais , Receptores de Apelina/metabolismo , Depressão/etiologia , Masculino , Vias Neurais/metabolismo , Fenótipo , Ratos Sprague-Dawley , Restrição Física , Estresse Psicológico/complicações , Regulação para Cima
9.
Artigo em Inglês | MEDLINE | ID: mdl-30150475

RESUMO

Voriconazole is a broad-spectrum triazole antifungal and the first-line treatment for invasive aspergillosis (IA). The aim of this research was to study the dose adjustments of voriconazole as well as the affecting factors influencing voriconazole trough concentrations in Asian children to optimize its daily administration. Clinical data were analyzed of inpatients 2 to 14 years old who were subjected to voriconazole trough concentration monitoring from 1 June 2015 to 1 December 2017. A total of 138 voriconazole trough concentrations from 42 pediatric patients were included. Voriconazole trough concentrations at steady state ranged from 0.02 to 9.35 mg/liter, with high inter- and intraindividual variability. Only 50.0% of children achieved the target range (1.0 to 5.5 mg/liter) at initial dosing, while 35.7% of children were subtherapeutic, and 14.3% of children were supratherapeutic at initial dosing. There was no correlation between initial trough concentrations and initial dosing. A total of 28.6% of children (12/42) received an adjusted dose according to trough concentrations. Children <6, 6 to 12, and >12 years old required a median oral maintenance dose to achieve the target range of 11.1, 7.2, and 5.3 mg/kg twice daily, respectively (P = 0.043). The average doses required to achieved the target range were 7.7 mg/kg and 5.6 mg/kg, respectively, and were lower than the recommended dosage (P = 0.033 and 0.003, respectively). Affecting factors such as administration routes and coadministration with proton pump inhibitors (PPIs) explained 55.3% of the variability in voriconazole exposure. Therapeutic drug monitoring (TDM) of voriconazole could help to individualize antifungal therapy for children and provide guidelines for TDM and dosing optimization in Asian children.


Assuntos
Antifúngicos/farmacocinética , Aspergilose/tratamento farmacológico , Monitoramento de Medicamentos , Micoses/tratamento farmacológico , Voriconazol/farmacocinética , Adolescente , Antifúngicos/sangue , Antifúngicos/farmacologia , Aspergilose/sangue , Aspergilose/microbiologia , Aspergilose/patologia , Criança , Pré-Escolar , China , Esquema de Medicação , Cálculos da Dosagem de Medicamento , Feminino , Humanos , Lactente , Masculino , Micoses/sangue , Micoses/microbiologia , Micoses/patologia , Centros de Atenção Terciária , Voriconazol/sangue , Voriconazol/farmacologia
10.
ACS Omega ; 1(4): 696-705, 2016 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-31457157

RESUMO

Two types of out-of-substrate Ag-Ag2O nanoplates were grown on a ZnO substrate through a surfactantless photochemical method. First, the in situ photochemically synthesized Ag-Ag2O nanoparticles further crystallized into nanoplate-like superstructures with rough surfaces and ragged edges. The nanoparticle-mediated crystallization process was governed by a layer-by-layer crystallization mechanism. Our study should help fundamentally understand the formation mechanism of hierarchical nanoparticle superstructures. Under continuous UV illumination, the hundreds of nanometer-sized rough nanoplates (i.e., the nanoplate-like superstructures of nanoparticles) can be transformed into large smooth nanoplates with sizes of up to several micrometers. The out-of-substrate Ag-Ag2O nanoplates/ZnO heterostructures are potentially promising for photocatalytic applications.

11.
Oncotarget ; 6(38): 40667-79, 2015 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-26512779

RESUMO

The gelatinase inhibitor doxycycline is the prototypical antitumor antibiotic. We investigated the effects of doxycycline on the migration, invasion, and metastasis of human lung cancer cell lines and in a mouse model. We also measured the effect of doxycycline on the transcription of epithelial-mesenchymal transition (EMT) markers, and used immunohistochemistry to determine whether EMT reversal was associated with doxycycline inhibition. Doxycycline dose-dependently inhibited proliferation, migration, and invasion of NCI-H446 human small cell lung cancer cells. It also suppressed tumor growth from NCI-H446 and A549 lung cancer cell xenografts without altering body weight, inhibited Lewis lung carcinoma cell migration, and prolonged survival. The activities of the transcription factors Twist1/2, SNAI1/2, AP1, NF-κB, and Stat3 were suppressed by doxycycline, which reversed EMT and inhibited signal transduction, thereby suppressing tumor growth and metastasis. Our data demonstrate functional targeting of transcription factors by doxycycline to reverse EMT and suppress tumor proliferation and metastasis. Thus, doxycycline selectively targets malignant tumors and reduces its metastatic potential with less cytotoxicity in lung cancer patients.


Assuntos
Antibacterianos/farmacologia , Carcinoma Pulmonar de Lewis/prevenção & controle , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Doxiciclina/farmacologia , Neoplasias Pulmonares/prevenção & controle , Animais , Apoptose/efeitos dos fármacos , Western Blotting , Carcinoma Pulmonar de Lewis/metabolismo , Carcinoma Pulmonar de Lewis/patologia , Transição Epitelial-Mesenquimal , Imunofluorescência , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Técnicas Imunoenzimáticas , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Invasividade Neoplásica , Células Tumorais Cultivadas , Cicatrização/efeitos dos fármacos , Ensaios Antitumorais Modelo de Xenoenxerto
12.
FEMS Microbiol Lett ; 334(1): 27-34, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22697582

RESUMO

Phytophthora sojae is a devastating pathogen that causes soybean Phytophthora root rot. This study reports the development of a loop-mediated isothermal amplification (LAMP) assay targeting the A3aPro element for visual detection of P. sojae. The A3aPro-LAMP assay efficiently amplified the target element in < 80 min at 64 °C and was evaluated for specificity and sensitivity. The specificity was evaluated against P. sojae, Phytophthora spp., Pythium spp., and true fungi isolates. Magnesium pyrophosphate resulting from the LAMP of P. sojae could be detected by real-time measurement of turbidity. Phytophthora sojae DNA products were visualized as a ladder-like banding pattern on 2% gel electrophoresis. A positive colour (sky blue) was only observed in the presence of P. sojae with the addition of hydroxynaphthol blue prior to amplification, whereas none of other isolates showed a colour change. The detection limit of the A3aPro-specific LAMP assay for P. sojae was 10 pg µL(-1) of genomic DNA per reaction. The assay also detected P. sojae from diseased soybean tissues and residues. These results suggest that the A3aPro-LAMP assay reported here can be used for the visual detection of P. sojae in plants and production fields.


Assuntos
Glycine max/microbiologia , Técnicas de Amplificação de Ácido Nucleico/métodos , Phytophthora/isolamento & purificação , Doenças das Plantas/microbiologia , Primers do DNA/genética , Phytophthora/classificação , Phytophthora/genética
13.
J Biotechnol ; 150(1): 182-9, 2010 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-20691226

RESUMO

Restoration of lymphatic drainage using lymph vessels or tissue grafting is becoming an efficient method for alleviating obstructive lymphedema. However, the lack of ideal lymphatic grafts is the key problem that limits the application of lymphatic transplantation, but now that may be resolved with tissue-engineered lymph vessels. In this study, the feasibility of reconstructing lymph vessels was explored using lymphatic endothelial cells (LECs) combined with polyglycolic acid (PGA) scaffolds. The highly purified human dermal LECs can be isolated from human dermis by immunomagnetic bead sorting and multiplied in culture. The viability and growth potential of subcultured LECs make it possible to obtain large amount of cells in vitro. Light and scanning electron microscopy (SEM) showed that the prefabricated PGA scaffolds, with three-dimensional structure, can support cell adhesion, growth and spreading. The constructs formed with LECs combined with PGA scaffolds were cultured in vitro for 10 days and then implanted subcutaneously into nude mice. Six weeks after implantation, the portions of implanted tubules were harvested. Gross and histological observation demonstrated that the tubular structure still remained in the experimental groups but not in the control groups. Immunohistochemical staining and RT-PCR assay of the implanted vessels revealed positive staining in experimental groups for the lymphatic specific markers Podoplanin, VEGFR-3 and LYVE-1. The results indicate that LECs can serve as seed cells and be successfully combined with PGA scaffolds, and the tissue-engineered tubular structure using implanted LECs-PGA compounds showed preliminary characteristics of lymph vessels. A gap between the nearly normal or functional lymph vessel still exists as we have only the endothelial cell-lined duct, but this study demonstrates that it is feasible to construct tissue-engineered lymph vessels using LECs combined with a biodegradable material.


Assuntos
Prótese Vascular , Células Endoteliais/citologia , Vasos Linfáticos/citologia , Engenharia Tecidual/métodos , Alicerces Teciduais , Animais , Sobrevivência Celular , Prepúcio do Pênis/citologia , Histocitoquímica , Humanos , Masculino , Camundongos , Camundongos Nus , Microscopia Eletrônica de Varredura , Projetos Piloto , Ácido Poliglicólico/química , Reação em Cadeia da Polimerase Via Transcriptase Reversa
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