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1.
Clin Cosmet Investig Dermatol ; 17: 147-158, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38283796

RESUMO

Purpose: Vitiligo is an autoimmune disease that results in the loss of epidermal melanocytes. The treatments for patients with vitiligo remain lacking. Erzhiwan (EZW), a traditional Chinese Medicine composed of Ligustri Lucidi Fructus and Ecliptae Herba, was used to ameliorate depigmentation since ancient China. This study aims to investigate the effect of EZW on vitiligo-related depigmentation. Methods: A vitiligo-related depigmentation mouse model was induced by monobenzone and restraint stress. The experimental depigmentation mice were treated with EZW. Histological observation of skin was conducted. Cutaneous oxidative damage and inflammation were determined. A network pharmacology analysis was carried out. Results: EZW reduced depigmentation score (p<0.01), cutaneous inflammatory infiltration (p<0.01), and CD8α-positive expression (p<0.01), and increased cutaneous melanin content in experimental depigmentation mice. EZW reduced stress reaction in experimental depigmentation mice (p<0.01). EZW inhibited 8-hydroxy-2-deoxyguanosine (8-OHdG)-related DNA oxidative damage in the skin (p<0.05, p<0.01). In addition, EZW reduced cutaneous macrophage migration inhibitory factor (MIF)-CD74-NF-κB signaling (p<0.01). The network pharmacology analysis demonstrated that EZW regulated necroptosis, apoptosis, and FoxO signaling pathways in vitiligo. An in vitro experiment showed that the main ingredient of EZW, specnuezhenide, protected against monobenzone and MIF-induced cell death in HaCaT cells (p<0.01). Conclusion: EZW ameliorates restraint stress- and monobenzone-induced depigmentation via the inhibition of MIF and 8-OHdG signaling. The findings provide a data basis of an utilization of EZW in vitiligo.

2.
Acta Biochim Biophys Sin (Shanghai) ; 55(12): 1961-1971, 2023 12 25.
Artigo em Inglês | MEDLINE | ID: mdl-37997375

RESUMO

Psychological stress promotes nonalcoholic steatohepatitis (NASH) development. However, the pathogenesis of psychological stress-induced NASH remains unclear. This study aims to explore the underlying mechanism of restraint stress-induced NASH, which mimics psychological stress, and to discover potential NASH candidates. Methionine choline deficient diet- and high fat diet-induced hepatosteatotic mice are subjected to restraint stress to induce NASH. The mice are administrated with Xiaoyaosan granules, NOD-like receptor family pyrin domain containing 3 (NLRP3) inhibitors, farnesoid X receptor (FXR) agonists, or macrophage scavengers. Pathological changes and NLRP3 signaling in the liver are determined. These results demonstrate that restraint stress promotes hepatic inflammation and fibrosis in hepatosteatotic mice. Restraint stress increases the expressions of NLRP3, Caspase-1, Gasdermin D, interleukin-1ß, cholesterol 7α-hydroxylase, and sterol 12α-hydroxylase and decreases the expression of FXR in NASH mice. Xiaoyaosan granules reverse hepatic inflammation and fibrosis and target FXR and NLRP3 signals. In addition, inhibition of NLRP3 reduces the NLRP3 inflammasome and liver damage in mice with restraint stress-induced NASH. Elimination of macrophages and activation of FXR also attenuate inflammation and fibrosis by inhibiting NLRP3 signaling. However, NLRP3 inhibitors or macrophage scavengers fail to affect the expression of FXR. In conclusion, restraint stress promotes NASH-related inflammation and fibrosis by regulating the FXR/NLRP3 signaling pathway. Xiaoyaosan granules, NLRP3 inhibitors, FXR agonists, and macrophage scavengers are potential candidates for the treatment of psychological stress-related NASH.


Assuntos
Hepatopatia Gordurosa não Alcoólica , Camundongos , Animais , Hepatopatia Gordurosa não Alcoólica/etiologia , Hepatopatia Gordurosa não Alcoólica/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Fígado/metabolismo , Inflamassomos/metabolismo , Transdução de Sinais , Inflamação/metabolismo , Fibrose , Camundongos Endogâmicos C57BL
3.
BMC Genomics ; 24(1): 618, 2023 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-37853336

RESUMO

BACKGROUND: Extravillous trophoblast cell (EVT) differentiation and its communication with maternal decidua especially the leading immune cell type natural killer (NK) cell are critical events for placentation. However, appropriate in vitro modelling system and regulatory programs of these two events are still lacking. Recent trophoblast organoid (TO) has advanced the molecular and mechanistic research in placentation. Here, we firstly generated the self-renewing TO from human placental villous and differentiated it into EVTs (EVT-TO) for investigating the differentiation events. We then co-cultured EVT-TO with freshly isolated decidual NKs for further study of cell communication. TO modelling of EVT differentiation as well as EVT interaction with dNK might cast new aspect for placentation research. RESULTS: Single-cell RNA sequencing (scRNA-seq) was applied for comprehensive characterization and molecular exploration of TOs modelling of EVT differentiation and interaction with dNKs. Multiple distinct trophoblast states and dNK subpopulations were identified, representing CTB, STB, EVT, dNK1/2/3 and dNKp. Lineage trajectory and Seurat mapping analysis identified the close resemblance of TO and EVT-TO with the human placenta characteristic. Transcription factors regulatory network analysis revealed the cell-type specific essential TFs for controlling EVT differentiation. CellphoneDB analysis predicted the ligand-receptor complexes in dNK-EVT-TO co-cultures, which relate to cytokines, immunomodulation and angiogenesis. EVT was known to affect the immune properties of dNK. Our study found out that on the other way around, dNKs could exert effects on EVT causing expression changes which are functionally important. CONCLUSION: Our study documented a single-cell atlas for TO and its applications on EVT differentiation and communications with dNKs, and thus provide methodology and novel research cues for future study of human placentation.


Assuntos
Placenta , Trofoblastos , Gravidez , Feminino , Humanos , Trofoblastos/metabolismo , Decídua/metabolismo , Diferenciação Celular , Organoides , Células Matadoras Naturais/metabolismo , Movimento Celular
4.
Front Immunol ; 14: 1130308, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37006248

RESUMO

The human placenta is a unique temporary organ with a mysterious immune tolerance. The formation of trophoblast organoids has advanced the study of placental development. HLA-G is uniquely expressed in the extravillous trophoblast (EVT) and has been linked to placental disorders. With older experimental methodologies, the role of HLA-G in trophoblast function beyond immunomodulation is still contested, as is its role during trophoblast differentiation. Organoid models incorporating CRISPR/Cas9 technology were used to examine the role of HLA-G in trophoblast function and differentiation. JEG-3 trophoblast organoids (JEG-3-ORGs) were established that highly expressed trophoblast representative markers and had the capacity to differentiate into EVT. CRISPR/Cas9 based on HLA-G knockout (KO) significantly altered the trophoblast immunomodulatory effect on the cytotoxicity of natural killer cells, as well as the trophoblast regulatory effect on HUVEC angiogenesis, but had no effect on the proliferation and invasion of JEG-3 cells and the formation of TB-ORGs. RNA-sequencing analysis further demonstrated that JEG-3 KO cells followed similar biological pathways as their wild-type counterparts during the formation of TB-ORGs. In addition, neither HLA-G KO nor the exogenous addition of HLA-G protein during EVT differentiation from JEG-3-ORGs altered the temporal expression of the known EVT marker genes. Based on the JEG-3 KO (disruption of exons 2 and 3) cell line and the TB-ORGs model, it was determined that HLA-G has a negligible effect on trophoblast invasion and differentiation. Despite this, JEG-3-ORG remains a valuable model for studying trophoblast differentiation.


Assuntos
Placenta , Trofoblastos , Gravidez , Feminino , Humanos , Trofoblastos/metabolismo , Placenta/metabolismo , Antígenos HLA-G/genética , Antígenos HLA-G/metabolismo , Linhagem Celular Tumoral , Organoides
5.
Mater Horiz ; 10(7): 2535-2541, 2023 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-37070656

RESUMO

The use of crystalline metal-organic complexes with definite structures as multilevel memories can enable explicit structure-property correlations, which is significant for designing the next generation of memories. Here, four Zn-polysulfide complexes with different degrees of conjugation have been fabricated as memory devices. ZnS6(L)2-based memories (L = pyridine and 3-methylpyridine) can exhibit only bipolar binary memory performances, but ZnS6(L)-based memories (L = 2,2'-bipyridine and 1,10-phenanthroline) illustrate non-volatile ternary memory performances with high ON2/ON1/OFF ratios (104.22/102.27/1 and 104.85/102.58/1) and ternary yields (74% and 78%). Their ON1 states stem from the packing adjustments of organic ligands upon the injection of carriers, and the ON2 states are a result of the ring-to-chain relaxation of S62- anions. The lower conjugated degrees in ZnS6(L)2 result in less compact packing; consequently, the adjacent S62- rings are too long to trigger the S62- relaxation. The deep structure-property correlation in this work provides a new strategy for implementing multilevel memory by triggering polysulfide relaxation based on the conjugated degree regulation of organic ligands.

6.
J Food Biochem ; 46(12): e14428, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36125796

RESUMO

Biochanin A (Bio-A), an isoflavone abundant in chickpeas, possesses hypoglycemic, hypolipidemic, and anti-inflammatory effects. However, whether Bio-A has antihepatosteatosis effect remains unclear. This study aimed to evaluate the antihepatosteatosis effect of Bio-A on oleate (OA)-treated hepatocytes, and explore the underlying mechanism. When incubated with OA for 24 h, HepG2 cells were treated with various concentrations of Bio-A for 24 h to obtain an optimal antihepatosteatosis dose. HepG2 cells were treated with the AMP-activated protein kinase (AMPK) inhibitor Compound C, or the sirtuin-3 (SIRT3) inhibitor 3-TYP, and incubated with 50 µM Bio-A. The results indicated that 12.6% of lipid content, particularly 11.0% of triglyceride content, and the expression of adipocyte differentiation-related protein were significantly decreased in Bio-A-treated hepatosteatosis cells, followed by an increase in the expression of Beclin 1, phosphorylation of Unc-51-like kinase 1 (ULK-1), the microtubule-associated protein 1 light chain 3 (LC3)-II/LC3-I ratio, and a decrease in expression of p62. The results indicated that Bio-A upregulated autophagosome formation and autophagy flux. In addition, Bio-A increased SIRT3 expression and AMPK phosphorylation in OA-treated HepG2 cells. Blockade of AMPK and SIRT3 blocked the antihepatosteatosis effect and ULK-1 activation by Bio-A. AMPK inhibition did not eliminate the activation of SIRT3 by Bio-A. AutoDock analysis demonstrated that interaction might exist between Bio-A and SIRT3. In conclusion, Bio-A reduced fat accumulation in OA-treated HepG2 cells by activating SIRT3/AMPK/ULK-1-mediated autophagy. The findings provide a theoretical basis for the effect of Bio-A on hepatic steatosis-related diseases. PRACTICAL APPLICATIONS: This study highlights the antihepatosteatosis effects of biochanin A (Bio-A) on oleate (OA)-treated hepatocytes. Bio-A, one of the isoflavones in Cicer arietinum Linn., possesses multiple bioactivities such as antiobesity, anti-inflammation, and hypoglycemic and hypolipidemic effects. This study provides a new application of Bio-A to treat hepatic steatosis, and revealed the underlying mechanism of Bio-A involved in the activation of the SIRT3/AMPK/ULK-1-mediated autophagy. The findings provide a theoretical basis for the application of Bio-A to hepatic steatosis-related diseases.


Assuntos
Fígado Gorduroso , Sirtuína 3 , Humanos , Proteínas Quinases Ativadas por AMP/genética , Proteínas Quinases Ativadas por AMP/farmacologia , Células Hep G2 , Ácido Oleico/farmacologia , Transdução de Sinais , Sirtuína 3/genética , Sirtuína 3/metabolismo , Sirtuína 3/farmacologia
7.
Asian J Androl ; 24(6): 607-614, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35381696

RESUMO

Idiopathic asthenozoospermia, a common factor in male infertility, is characterized by altered sperm motility function in fresh ejaculate. Although the ß-defensin 126 (DEFB126) protein is associated with asthenozoospermia, DEFB126 gene polymorphisms have not been extensively studied. Therefore, the association between DEFB126 gene polymorphisms and asthenozoospermia requires further investigation. Screening was performed by semen analysis, karyotype analysis, and Y microdeletion detection, and 102 fertile men and 106 men with asthenozoospermia in Chengdu, China, were selected for DEFB126 gene sequence analyses. Seven nucleotide mutations and two nucleotide deletions in the DEFB126 gene were detected. rs11467417 (317-318 del/del), rs11467497 (163-166 wt/del), c.152T>C, and c.227A>G were significantly different between the control and asthenozoospermia groups, likely representing high-risk genetic factors for asthenozoospermia among males. DEFB126 expression was not observed in sperm with rs11467497 homozygous deletion and was unstable in sperm with rs11467417 homozygous deletion. The rs11467497 four-nucleotide deletion leads to truncation of DEFB126 at the carboxy-terminus, and the rs11467417 binucleotide deletion produces a non-stop messenger RNA (mRNA). The above deletions may be responsible for male hypofertility and infertility by reducing DEFB126 affinity to sperm surfaces. Based on in silico analysis, the amino acids 51M and 76K are located in the highly conserved domain; c.152T>C (M51T) and c.227A>G (K76R) are predicted to be damaging and capable of changing alternative splice, structural and posttranslational modification sites of the RNA, as well as the secondary structure, structural stability, and hydrophobicity of the protein, suggesting that these mutations are associated with asthenozoospermia.


Assuntos
Astenozoospermia , beta-Defensinas , Masculino , Humanos , Astenozoospermia/genética , Astenozoospermia/metabolismo , Motilidade dos Espermatozoides/genética , Homozigoto , Polimorfismo de Nucleotídeo Único , Sêmen , Deleção de Sequência/genética , Espermatozoides/metabolismo , Nucleotídeos/metabolismo , beta-Defensinas/genética , beta-Defensinas/metabolismo
8.
Medicine (Baltimore) ; 101(9): e28960, 2022 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-35244058

RESUMO

BACKGROUND: Female breast cancer is the most common cancer nowadays, and its treatment has a significant impact on patients both physically and psychologically. Many randomized trials have proved that case management (CM) can effectively care for patients. However, there is a lack of systematic scientific evaluation, so this systematic evaluation aims to explore the impact of CM on breast cancer patients. METHODS: PubMed, Embase, Cochrane Library, Scopus, CINAHL were searched. Chinese repositories included China National Knowledge, Infrastructure Database (CNKI), Wan fang Database, China Biology Medicine Database. We will also search unpublished literature at ClinicalTrials.gov. Randomized controlled trials were collected from them. The literature will be screened according to inclusion and exclusion criteria, and 2 researchers will extract the literature independently. The primary outcome indicator for this study will be patient satisfaction. Statistics were performed using RevMan 5.4 software. The quality of each outcome will be evaluated using the Grading of Recommendations Assessment, Development, and Evaluation. RESULTS: This study will provide the most recent evidence for evaluating the impact of CM on breast cancer patients. CONCLUSION: To evaluate the impact of CM on patients with breast cancer. REGISTRATION NUMBER: DOI:10.17605/OSF.IO/ZJKHX.


Assuntos
Neoplasias da Mama/terapia , Administração de Caso , Feminino , Humanos , Metanálise como Assunto , Satisfação do Paciente , Projetos de Pesquisa , Revisões Sistemáticas como Assunto
9.
Plant Dis ; 2021 Feb 23.
Artigo em Inglês | MEDLINE | ID: mdl-33622060

RESUMO

Cornus hongkongensis (Hemsl.) is an excellent ornamental tree species in China and elsewhere. In 2019, C. hongkongensis anthracnose was firstly observed at the campus of Jiangxi Agricultural University (JXAU) (28°45'56″N, 115°50'21″E), then found in parks, Nanchang, China. In early August, the disease appeared and lasted until the leaves dropped (November). The disease incidence was above 60%, and the diseased leaf rate was above 70%. The lesions mostly appeared along the leaf edges. Some small round to irregular lesions also developed in other parts of the leaves. These diseased leaves had circular or irregularly shaped spots with gray-white color in the center and dark brown on the edge of the lesions. Later, the lesions became necrotic and shriveled. As the disease progressed, the spots coalesced so that affected leaves appeared blighted (Supplementary Figure 1 A-C). To identify the pathogen, leaves with typical symptoms from the campus of JXAU were collected and small pieces (5 × 5 mm) from the lesion borders were surfaced sterilized in 70% ethanol for 30 s, followed by 1 min in 3% NaOCl, and then rinsed with sterile distilled water three times. Leaf pieces were placed on potato dextrose agar (PDA) and incubated at 25 °C under a 12-h light/dark cycle (3000 lx). Pure cultures were obtained from individual conidia by single spore isolates. For studies of microscopic morphology, a representative isolate JX-S4 was subcultured on PDA. The colony of JX-S4 was white and turning gray and light gray on the reverse side, producing dark-green pigmentation near the center (Supplementary Figure 1 D). The conidia were one-celled, straight, hyaline, subcylindrical with rounded ends and 16.9 ± 1.6 × 6.0 ± 0.6 µm (n = 50) in size. Appressoria were one-celled, pale brown, thick-walled, ellipsoidal, and measured 8.7 ± 1.7 × 6.4 ± 0.8 µm (n = 50) (Supplementary Figure 1 E, F). The morphological characteristics of JX-S4 matched those of the Colletotrichum siamense species (Weir et al. 2012). For accurate identification, the internal transcribed spacer (ITS) and the genes encoding glyceraldehyde-3-phosphate dehydrogenase (GAPDH), chitin synthase (CHS-I), beta-tubulin 2 (TUB2), and calmodulin (CAL) were respectively amplified with primers ITS1/ITS4, GDF/GDR, CHS-79F/CHS-345R, ßt2a/ßt2b, and CL1/CL2. The sequences were deposited in GenBank (Accession nos. MT587807, MT628710, MT628709, MT628711, and MT628708). Phylogenetic analysis was calculated with concatenated sequences (ITS, GAPDH, CHS-I, CAL, and TUB2) using MEGA 7. In the maximum likelihood phylogenetic tree, Isolate JX-S4 was clustered with C. siamense with 93% bootstrap support (Supplementary Figure 2). Based on the morphological characteristics and phylogenetic analysis, JX-S4 was identified as C. siamense. Pathogenicity test of JX-S4 was verified on 45 attached healthy leaves from three C. hongkongensis plants (10-year-old) at the campus of JXAU inoculated with mycelial plugs (φ=5 mm) from the culture edge (6-day-old) on PDA. And an additional 45 healthy leaves were inoculated with PDA plugs as controls. The leaves were wounded with a red-hot needle (φ=0.5 mm). All treatment and control leaves were wrapped up with black plastic bags to keep them moist for 2 days. The pathogenicity tests were repeated twice. Within 7 days, all the inoculated leaves developed the lesions, which were similar to those observed in the field. Control leaves were asymptomatic (Supplementary Figure 1 G, H). The same fungus was re-isolated from the symptomatic tissues, fulfilling Koch's postulates. To our knowledge, this is the first report of C. siamense causing C. hongkongensis anthracnose. This finding provides crucial information for managing this disease. For example, when diagnosing Cornus anthracnose, C. siamense needs to be looked out for and appropriate control measures implemented.

10.
Mol Plant ; 13(6): 879-893, 2020 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-32298785

RESUMO

In response to far-red light (FR), FAR-RED ELONGATED HYPOCOTYL 1 (FHY1) transports the photoactivated phytochrome A (phyA), the primary FR photoreceptor, into the nucleus, where it initiates FR signaling in plants. Light promotes the 26S proteasome-mediated degradation of FHY1, which desensitizes FR signaling, but the underlying regulatory mechanism remains largely unknown. Here, we show that reversible SUMOylation of FHY1 tightly regulates this process. Lysine K32 (K32) and K103 are major SUMOylation sites of FHY1. We found that FR exposure promotes the SUMOylation of FHY1, which accelerates its degradation. Furthermore, we discovered that ARABIDOPSIS SUMO PROTEASE 1 (ASP1) interacts with FHY1 in the nucleus under FR and facilitates its deSUMOylation. FHY1 was strongly SUMOylated and its protein level was decreased in the asp1-1 loss-of-function mutant compared with that in the wild type under FR. Consistently, asp1-1 seedlings exhibited a decreased sensitivity to FR, suggesting that ASP1 plays an important role in the maintenance of proper FHY1 levels under FR. Genetic analysis further revealed that ASP1 regulates FR signaling through an FHY1- and phyA-dependent pathway. Interestingly, We found that continuous FR inhibits ASP1 accumulation, perhaps contributing to the desensitization of FR signaling. Taken together, these results indicate that FR-induced SUMOylation and ASP1-dependent deSUMOylation of FHY1 represent a key regulatory mechanism that fine-tunes FR signaling.


Assuntos
Proteínas de Arabidopsis/metabolismo , Fitocromo A/metabolismo , Fitocromo/metabolismo , Transdução de Sinais , Sumoilação , Luz , Modelos Biológicos , Ligação Proteica , Estabilidade Proteica/efeitos da radiação , Proteólise/efeitos da radiação , Proteínas Modificadoras Pequenas Relacionadas à Ubiquitina/metabolismo , Especificidade por Substrato
11.
Arch Insect Biochem Physiol ; 102(3): e21593, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31612553

RESUMO

The diamondback moth, Plutella xylostella, is one of the most destructive pests worldwide and its management relies exclusively on frequent application of chemical insecticides. Resistance to common insecticides is now widespread, and novel classes of insecticides are needed. Entomopathogenic bacteria and their related products play an important role in the management of this pest. In the present work, one bacterial strain was separated from infected pupae of P. xylostella collected from field and its pathogenicity was evaluated. On the basis of the 16S ribosomal RNA sequencing, BLASTN, and phylogenetic analysis, this bacterial isolate was identified as Pseudomonas cedrina. Oral administration of P. cedrina at levels above 10,000 CFU/ml gave significant mortality to P. xylostella larvae. The pathogenicity was also observed by reduced longevity and fecundity in adult females. However, when live bacterial cells were removed, the cultured broth lost any pathogenicity. In response to the bacterial infection, P. xylostella expressed antimicrobial and stress-associated genes. A mixture treatment of P. cedrina and Bacillus thuringiensis showed an additive effect on larval mortality of P. xylostella. These results indicated that P. cedrina is an opportunistic entomopathogen without secretion of toxins. Furthermore, the additive effect of P. cedrina and B. thuringiensis provide a new insight to develop new strategy for controlling P. xylostella.


Assuntos
Mariposas/microbiologia , Pseudomonas/isolamento & purificação , Animais , Feminino , Fertilidade , Perfilação da Expressão Gênica , Larva/microbiologia , Longevidade , Mariposas/genética , Mariposas/metabolismo , Controle Biológico de Vetores/métodos , Filogenia , Pseudomonas/classificação , Pseudomonas/patogenicidade , Pupa/microbiologia , RNA Ribossômico 16S
12.
Mol Plant ; 12(2): 215-228, 2019 02 04.
Artigo em Inglês | MEDLINE | ID: mdl-30543996

RESUMO

Plant immune responses are tightly regulated to ensure their appropriate deployment. Overexpression of TOPLESS-RELATED 1 (TPR1), a SUPPRESSOR OF npr1-1, CONSTITUTIVE 1 (SNC1)-interacting protein, results in autoimmunity that reduces plant growth and development. However, how TPR1 activity is regulated remains unknown. Loss of function of SIZ1, a (SUMO) E3 ligase, induces an autoimmune response, partially due to elevated SNC1 levels. Here we show that SNC1 expression is upregulated in Arabidopsis thaliana siz1-2 due to positive-feedback regulation by salicylic acid. SIZ1 physically interacts with TPR1 and facilitates its SUMO modification. The K282 and K721 residues in TPR1 serve as critical SUMO attachment sites. Simultaneous introduction of K282R and K721R substitutions in TPR1 blocked its SUMOylation, enhanced its transcriptional co-repressor activity, and increased its association with HISTONE DEACETYLASE 19 (HDA19), suggesting that SUMOylation of TPR1 represses its transcriptional co-repressor activity and inhibits its interaction with HDA19. In agreement with this finding, the simultaneous introduction of K282R and K721R substitutions enhanced TPR1-mediated immunity, and the tpr1 mutation partially suppressed autoimmunity in siz1-2. These results demonstrate that SIZ1-mediated SUMOylation of TPR1 represses plant immunity, which at least partly contributes to the suppression of autoimmunity under non-pathogenic conditions to ensure proper plant development.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/imunologia , Arabidopsis/metabolismo , Ligases/metabolismo , Imunidade Vegetal , Sumoilação , Substituição de Aminoácidos , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Proteínas de Neoplasias/genética , Transcrição Gênica
13.
PLoS Genet ; 12(4): e1006016, 2016 04.
Artigo em Inglês | MEDLINE | ID: mdl-27128446

RESUMO

COP1 (CONSTITUTIVE PHOTOMORPHOGENIC 1), a ubiquitin E3 ligase, is a central negative regulator of photomorphogenesis. However, how COP1 activity is regulated by post-translational modifications remains largely unknown. Here we show that SUMO (small ubiquitin-like modifier) modification enhances COP1 activity. Loss-of-function siz1 mutant seedlings exhibit a weak constitutive photomorphogenic phenotype. SIZ1 physically interacts with COP1 and mediates the sumoylation of COP1. A K193R substitution in COP1 blocks its SUMO modification and reduces COP1 activity in vitro and in planta. Consistently, COP1 activity is reduced in siz1 and the level of HY5, a COP1 target protein, is increased in siz1. Sumoylated COP1 may exhibits higher transubiquitination activity than does non-sumoylated COP1, but SIZ1-mediated SUMO modification does not affect COP1 dimerization, COP1-HY5 interaction, and nuclear accumulation of COP1. Interestingly, prolonged light exposure reduces the sumoylation level of COP1, and COP1 mediates the ubiquitination and degradation of SIZ1. These regulatory mechanisms may maintain the homeostasis of COP1 activity, ensuing proper photomorphogenic development in changing light environment. Our genetic and biochemical studies identify a function for SIZ1 in photomorphogenesis and reveal a novel SUMO-regulated ubiquitin ligase, COP1, in plants.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Ligases/genética , Desenvolvimento Vegetal/genética , Ubiquitina-Proteína Ligases/genética , Substituição de Aminoácidos/genética , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/metabolismo , Fatores de Transcrição de Zíper de Leucina Básica/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Regulação da Expressão Gênica de Plantas , Ligases/metabolismo , Luz , Mutação , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Proteólise , Plântula/genética , Plântula/crescimento & desenvolvimento , Proteínas Modificadoras Pequenas Relacionadas à Ubiquitina/genética , Proteínas Modificadoras Pequenas Relacionadas à Ubiquitina/metabolismo , Sumoilação/genética , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitinação/genética
14.
Insect Sci ; 22(3): 375-85, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26013400

RESUMO

Microbial abundance and diversity of different life stages (fourth instar larvae, pupae and adults) of the diamondback moth, Plutella xylostella L., collected from field and reared in laboratory, were investigated using bacteria culture-dependent method and PCR-DGGE analysis based on the sequence of bacteria 16S rRNA V3 region gene. A large quantity of bacteria was found in all life stages of P. xylostella. Field population had higher quantity of bacteria than laboratory population, and larval gut had higher quantity than pupae and adults. Culturable bacteria differed in different life stages of P. xylostella. Twenty-five different bacterial strains were identified in total, among them 20 strains were presented in larval gut, only 8 strains in pupae and 14 strains in adults were detected. Firmicutes bacteria, Bacillus sp., were the most dominant species in every life stage. 15 distinct bands were obtained from DGGE electrophoresis gel. The sequences blasted in GenBank database showed these bacteria belonged to six different genera. Phylogenetic analysis showed the sequences of the bacteria belonged to the Actinobacteri, Proteobacteria and Firmicutes. Serratia sp. in Proteobacteria was the most abundant species in larval gut. In pupae, unculturable bacteria were the most dominant species, and unculturable bacteria and Serratia sp. were the most dominant species in adults. Our study suggested that a combination of molecular and traditional culturing methods can be effectively used to analyze and to determine the diversity of gut microflora. These known bacteria may play important roles in development of P. xylostella.


Assuntos
Bactérias/isolamento & purificação , Mariposas/microbiologia , Animais , Bactérias/genética , Contagem de Colônia Microbiana , DNA Bacteriano/análise , Trato Gastrointestinal/microbiologia , Larva/microbiologia , Mariposas/crescimento & desenvolvimento , Filogenia , Pupa/microbiologia , RNA Ribossômico 16S/análise
15.
Insect Sci ; 22(5): 619-28, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25183343

RESUMO

Larvae of the diamondback moth, Plutella xylostella L. (Lepidoptera: Plutellidae), have rich microbial communities inhabiting the gut, and these bacteria contribute to the fitness of the pest. In this study we evaluated the effects of five antibiotics (rifampicin, ampicillin, tetracycline, streptomycin sulfate and chloramphenicol) on the gut bacterial diversity of P. xylostella larvae. We screened five different concentrations for each antibiotic in a leaf disc assay, and found that rifampicin and streptomycin sulfate at 3 mg/mL significantly reduced the diversity of the bacterial community, and some bacterial species could be rapidly eliminated. The number of gut bacteria in the rifampicin group and streptomycin sulfate group decreased more rapidly than the others. With the increase of antibiotic concentration, the removal efficiency was improved, whereas toxic effects became more apparent. All antibiotics reduced larval growth and development, and eventually caused high mortality, malformation of the prepupae, and hindered pupation and adult emergence. Among the five antibiotics, tetracycline was the most toxic and streptomycin sulfate was a relatively mild one. Some dominant bacteria were not affected by feeding antibiotics alone. Denaturing gradient gel electrophoresis graph showed that the most abundant and diverse bacteria in P. xylostella larval gut appeared in the cabbage feeding group, and diet change and antibiotics intake influenced gut flora abundance. Species diversity was significantly reduced in the artificial diet and antibiotics treatment groups. After feeding on the artificial diet with rifampicin, streptomycin sulfate and their mixture for 10 days, larval gut bacteria could not be completely removed as detected with the agarose gel electrophoresis method.


Assuntos
Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Mariposas/efeitos dos fármacos , Animais , Bactérias/isolamento & purificação , Biodiversidade , Brassica , Eletroforese em Gel de Gradiente Desnaturante , Dieta , Trato Gastrointestinal/microbiologia , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Larva/microbiologia , Mariposas/crescimento & desenvolvimento , Mariposas/microbiologia
16.
J Alzheimers Dis ; 40(1): 153-9, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24346218

RESUMO

BACKGROUND: Alzheimer-associated neuronal thread protein (AD7c-NTP) has been reported to have high diagnostic accuracy in patients with Alzheimer's disease (AD). OBJECTIVE: To determine the diagnostic accuracy of urinary AD7c-NTP for the diagnosis of AD in patients with suspected AD. METHODS: We searched MEDLINE (January 1950 to date) and other electronic databases (from inception to date) for diagnostic accuracy studies that compared urinary AD7c-NTP to the standard clinical diagnosis of AD. We conducted citation searches and screened the reference lists of included studies. Studies were assessed for methodological quality using QUADAS. Summary receiver operating characteristic curves were used to summarize overall test performance. RESULT: Nine studies met our inclusion criteria. The summary estimates of the urinary AD7c-NTP assay for probable or possible AD were as follows: SEN, 0.87 (95%CI: 0.80-0.91); SPE, 0.89 (95%CI: 0.87-0.91); PLR, 8.13 (95% CI: 6.60-10.02); and NLR, 0.15 (95% CI: 0.10-0.22). The four summary estimates of urinary AD7c-NTP assay for probable AD were 0.89 (95% CI: 0.86-0.92), 0.90 (95% CI: 0.88-0.92), 8.88 (95% CI: 7.09-11.12), and 0.12 (95% CI: 0.09-0.16), with no obvious heterogeneity. CONCLUSION: Urinary AD7c-NTP is a sensitive and specific test for the diagnosis of probable AD. However, whether urinary AD7c-NTP can be used as an early marker is still unknown.


Assuntos
Doença de Alzheimer/diagnóstico , Doença de Alzheimer/urina , Proteínas do Tecido Nervoso/urina , Humanos , MEDLINE/estatística & dados numéricos , Editoração/estatística & dados numéricos , Curva ROC
17.
Neurol Sci ; 34(11): 1887-92, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24036689

RESUMO

Serum glial fibrillary acidic protein (GFAP) has been reported to have high diagnosis accuracy for differentiating intracerebral hemorrhage (ICH) from ischemic stroke (IS) in patients within acute phase of stroke symptom onset. Our purpose was to perform a systematic review and diagnostic meta-analysis to evaluate the valuation of serum GFAP in the early identification of ICH and IS. We searched MEDLINE, EMBASE and other electronic databases for diagnostic accuracy studies that compared serum GFAP with standard clinical diagnosis of ICH and IS in patients with symptoms of acute stroke. All publication years were included through to April 2013. The sensitivity (SEN), specificity (SPE), and positive and negative likelihood ratios (PLR and NLR, respectively) of serum GFAP for differentiating ICH and IS were pooled using a bivariate meta-analysis. Summary receiver operating characteristic curves were used to summarize overall test performance. A total of five trials met our inclusion criteria. The summarized estimates of serum GFAP for the differentiation of ICH and IS within 24 h of symptom onset were as follows: SEN, 81.1% (95% CI, 72.6-87.5%); SPE, 95.2% (95% CI 82.1-98.9%); PLR, 16.945 (95% CI 4.173-68.803); NLR, 0.198 (95% CI 0.133-0.296), significant heterogeneity was present. The four summary estimates of serum GFAP for patients within 1-6 h of symptom onset were 81.1% (95% CI 72.5-88.0%), 97.0% (95% CI 94.3-98.4%), 26.786 (95% CI 13.979-51.324), 0.191 (95% CI 0.126-0.291), respectively, with no obvious heterogeneity. Serum GFAP is a sensitive and specific test for differentiating ICH and IS in patients within 1-6 h of acute stroke symptom onset.


Assuntos
Hemorragia Cerebral/diagnóstico , Proteína Glial Fibrilar Ácida/sangue , Acidente Vascular Cerebral/diagnóstico , Doença Aguda , Biomarcadores/sangue , Hemorragia Cerebral/sangue , Humanos , Acidente Vascular Cerebral/sangue
18.
Plant Physiol ; 162(2): 1030-41, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23656895

RESUMO

Arabidopsis (Arabidopsis thaliana) CYCLIN-DEPENDENT KINASE Ds (CDKDs) phosphorylate the C-terminal domain of the largest subunit of RNA polymerase II. Arabidopsis CYCLIN H;1 (CYCH;1) interacts with and activates CDKDs; however, the physiological function of CYCH;1 has not been determined. Here, we report that CYCH;1, which is localized to the nucleus, positively regulates blue light-induced stomatal opening. Reduced-function cych;1 RNA interference (cych;1 RNAi) plants exhibited a drought tolerance phenotype. CYCH;1 is predominantly expressed in guard cells, and its expression was substantially down-regulated by dehydration. Transpiration of intact leaves was reduced in cych;1 RNAi plants compared with the wild-type control in light but not in darkness. CYCH;1 down-regulation impaired blue light-induced stomatal opening but did not affect guard cell development or abscisic acid-mediated stomatal closure. Microarray and real-time polymerase chain reaction analyses indicated that CYCH;1 did not regulate the expression of abscisic acid-responsive genes or light-induced stomatal opening signaling determinants, such as MYB60, MYB61, Hypersensitive to red and blue1, and Protein phosphatase7. CYCH;1 down-regulation induced the expression of redox homeostasis genes, such as LIPOXYGENASE3 (LOX3), LOX4, ARABIDOPSIS GLUTATHIONE PEROXIDASE 7 (ATGPX7), EARLY LIGHT-INDUCIBLE PROTEIN1 (ELIP1), and ELIP2, and increased hydrogen peroxide production in guard cells. Furthermore, loss-of-function mutations in CDKD;2 or CDKD;3 did not affect responsiveness to drought stress, suggesting that CYCH;1 regulates the drought stress response in a CDKD-independent manner. We propose that CYCH;1 regulates blue light-mediated stomatal opening by controlling reactive oxygen species homeostasis.


Assuntos
Arabidopsis/fisiologia , Ciclina H/metabolismo , Estômatos de Plantas/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Estresse Fisiológico , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Ciclina H/genética , Quinases Ciclina-Dependentes/genética , Quinases Ciclina-Dependentes/metabolismo , Secas , Regulação da Expressão Gênica de Plantas , Luz , Mutação , Transpiração Vegetal , Plantas Geneticamente Modificadas , Interferência de RNA
19.
Zhonghua Fu Chan Ke Za Zhi ; 46(7): 496-500, 2011 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-22041440

RESUMO

OBJECTIVE: To investigate the relationship between vaginal and intestinal candida in patients with vulvovaginal candidiasis by using microbiological and molecular methods. METHODS: The samples of vaginal discharge and anal swabs were collected from 148 cases with vulvovaginal candidiasis, followed by fungal culture, identification, purification and genome DNA extraction. The genome sequences from respective locations were aligned and typed according to their homology analyzed by internal transcribed spacer (ITS) PCR and random amplified polymorphic DNA (RAPD) PCR. Patients with vulvovaginal infection or those with infections in intestine and vulvovagina were pooled respectively, while the recurrent incidences after local anti-fungal treatments were analyzed. RESULTS: Candida albicans is the dominant pathogen in 148 cases with vulvovaginal candidiasis (91.9%, 136/148); 33.1% (49/148) of patients with vulvovaginal candidiasis were infected in both intestine and vulvovagina. While 92% (22/24) of patients with intestinal and vaginal candida infection showed high homology. The recurrent rate of patients with vulvovaginal candidiasis complicated with concurrent intestinal candida infection (7/14) was significantly higher than that of solo vaginal infected patients [21% (6/29)] after vaginal treatment (P<0.05). CONCLUSIONS: The infection of vulvovaginal candidiasis is highly associated with the concurrent infection of intestinal candida. The recurrent rate is high in patients with vulvovaginal candidiasis with concurrent infection of intestinal candida after vaginal treatment. The general management to those patients infected by both vulvovaginal and intestinal candida is necessary in reducing the recurrence of the disease.


Assuntos
Candida/genética , Candidíase Vulvovaginal/microbiologia , Intestinos/microbiologia , Reação em Cadeia da Polimerase/métodos , Vagina/microbiologia , Adolescente , Adulto , Antifúngicos/administração & dosagem , Antifúngicos/farmacologia , Candida/isolamento & purificação , Candidíase Vulvovaginal/complicações , Candidíase Vulvovaginal/epidemiologia , Portador Sadio , DNA Espaçador Ribossômico/genética , Feminino , Humanos , Enteropatias/complicações , Enteropatias/microbiologia , Pessoa de Meia-Idade , Técnica de Amplificação ao Acaso de DNA Polimórfico , Recidiva , Análise de Sequência de DNA , Adulto Jovem
20.
Zhonghua Xin Xue Guan Bing Za Zhi ; 39(5): 414-9, 2011 May.
Artigo em Chinês | MEDLINE | ID: mdl-21781595

RESUMO

OBJECTIVES: To isolate the cardiogenic fraction, which can enhance cardiogenic differentiation of bone marrow-derived mesenchymal stem cells (MSC) from Geum japonicum. The therapeutic effect of the isolated cardiogenic fraction was further tested in a rat myocardial infarction (MI) model. METHOD: Bioassay guided fractionation method was used for the isolation of the cardiogenic fraction, named as heart repair fraction (HRF). MI was induced by a permanent ligation of left anterior descending coronary artery. The rats exhibiting similarly decreased values of left ventricle ejection fraction (LVEF) and fraction shortening (LVFS) were used. The rats in test group (n = 10) were subject to HRF treatment (20 mg×kg(-1)×d(-1)) through gastric gavage daily for 4 weeks. Water alone (2 ml/d) was given through gastric gavage to rats in the control group (n = 10). The cardiac function was assessed by echocardiography at different time points. Masson trichrome staining was used for evaluation of the infarct size. Morphological and immunohistochemical studies were performed to investigate the HRF mediated myocardial regeneration. RESULTS: LVEF (66.2% ± 6.9%) and LVFS (46.8% ± 5.8%) were significantly increased two weeks post HRF treatment compared with the values (LVEF: 55.7% ± 6.0% and LVFS: 36.4% ± 5.2%) in control rats (all P < 0.01). The improved heart function was further restored 4 weeks post HRF treatment (P < 0.01). Furthermore, the treatment of acute MI with this HRF significantly reduced the infarct size (19.0% ± 6.1%) compared with that (31.1% ± 8.6%) in control rats (P < 0.01). Substantial regeneration of cardiomyocytes in infarcted region of the HRF treated heart was also observed that replaced a considerable part of the infarcted heart tissues resulting in remarkable reduction of the infarct size. CONCLUSION: The properties of this HRF isolated from Geum japonicum in stimulating substantial regeneration of myocardium in infarct region with consequently improved cardiac function appear to be new and represent a new approach for the treatment of MI.


Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Geum/química , Infarto do Miocárdio/tratamento farmacológico , Miócitos Cardíacos/efeitos dos fármacos , Animais , Masculino , Ratos , Ratos Sprague-Dawley , Regeneração
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