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1.
Acta Pharmacol Sin ; 44(10): 2103-2112, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37193754

RESUMO

Checkpoint inhibitors such as PD-1/PD-L1 antibody therapeutics are a promising option for the treatment of multiple cancers. Due to the inherent limitations of antibodies, great efforts have been devoted to developing small-molecule PD-1/PD-L1 signaling pathway inhibitors. In this study we established a high-throughput AlphaLISA assay to discover small molecules with new skeletons that could block PD-1/PD-L1 interaction. We screened a small-molecule library of 4169 compounds including natural products, FDA approved drugs and other synthetic compounds. Among the 8 potential hits, we found that cisplatin, a first-line chemotherapeutic drug, reduced AlphaLISA signal with an EC50 of 8.3 ± 2.2 µM. Furthermore, we showed that cisplatin-DMSO adduct, but not semplice cisplatin, inhibited PD-1/PD-L1 interaction. Thus, we assessed several commercial platinum (II) compounds, and found that bis(benzonitrile) dichloroplatinum (II) disturbed PD-1/PD-L1 interaction (EC50 = 13.2 ± 3.5 µM). Its inhibitory activity on PD-1/PD-L1 interaction was confirmed in co-immunoprecipitation and PD-1/PD-L1 signaling pathway blockade bioassays. Surface plasmon resonance assay revealed that bis(benzonitrile) dichloroplatinum (II) bound to PD-1 (KD = 2.08 µM) but not PD-L1. In immune-competent wild-type mice but not in immunodeficient nude mice, bis(benzonitrile) dichloroplatinum (II) (7.5 mg/kg, i.p., every 3 days) significantly suppressed the growth of MC38 colorectal cancer xenografts with increasing tumor-infiltrating T cells. These data highlight that platinum compounds are potential immune checkpoint inhibitors for the treatment of cancers.


Assuntos
Cisplatino , Inibidores de Checkpoint Imunológico , Neoplasias , Animais , Humanos , Camundongos , Anticorpos , Antígeno B7-H1/metabolismo , Cisplatino/farmacologia , Cisplatino/uso terapêutico , Camundongos Nus , Neoplasias/tratamento farmacológico , Receptor de Morte Celular Programada 1 , Inibidores de Checkpoint Imunológico/farmacologia
2.
J Enzyme Inhib Med Chem ; 38(1): 2244696, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37553905

RESUMO

A series of novel triazole derivatives containing aryl-propanamide side chains was designed and synthesised. In vitro antifungal activity studies demonstrated that most of the compounds inhibited the growth of six human pathogenic fungi. In particular, parts of phenyl-propionamide-containing compounds had excellent, broad-spectrum antifungal activity against Candida albicans SC5314, Cryptococcus neoformans 22-21, Candida glabrata 537 and Candida parapsilosis 22-20 with MIC values in the range of ≤0.125 µg/mL-4.0 µg/mL. In addition, compounds A1, A2, A6, A12 and A15 showed inhibitory activities against fluconazole-resistant Candida albicans and Candida auris. Preliminary structure-activity relationships (SARs) are also summarised. Moreover, GC-MS analysis demonstrated that A1, A3, and A9 interfered with the C. albicans ergosterol biosynthesis pathway by inhibiting Cyp51. Molecular docking studies elucidated the binding modes of A3 and A9 with Cyp51. These compounds with low haemolytic activity and favourable ADME/T properties are promising for the development of novel antifungal agents.


Assuntos
Antifúngicos , Triazóis , Humanos , Antifúngicos/química , Triazóis/química , Simulação de Acoplamento Molecular , Fluconazol/farmacologia , Candida albicans , Relação Estrutura-Atividade , Testes de Sensibilidade Microbiana
3.
Analyst ; 147(13): 3096-3100, 2022 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-35695068

RESUMO

Oxidized multi-walled carbon nanotube/nano-gold (AuNP-ox-MWCNT) composites with strong electrochemiluminescence (ECL) activity were applied to construct a new ECL immunosensor for the detection of carcinoembryonic antigen (CEA). The immunosensor showed a linear response range of 10-100 ng mL-1 and detection limit of 0.76 ng mL-1 (at a signal-to-noise ratio of 3). The as-developed immunosensor exhibited several advantages, including being simple to fabricate and being label free. The results indicated that ox-MWCNTs as a luminescent material have great application potential in analysis.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Nanotubos de Carbono , Técnicas Biossensoriais/métodos , Antígeno Carcinoembrionário/análise , Técnicas Eletroquímicas/métodos , Ouro , Imunoensaio , Limite de Detecção , Medições Luminescentes/métodos
4.
Bioorg Chem ; 129: 106216, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36283177

RESUMO

In order to develop new triazole derivatives, we optimized the lead compound a6 by structural modifications to obtain a series of (2R,3R)-3-((1-substituted-1H-1,2,3-triazol-4-yl) methoxy)-2-(2,4-difluorophenyl)-1-(1H-1,2,4-triazol-1-yl) butan-2-ol, compounds 5-36. Most of the target compounds exhibited excellent in vitro antifungal activity against Candida albicans 10231 and Candida glabrata 537 with MIC ≤ 0.125 µg/mL. Of particular note, compounds 6, 22, 28, 30 and 36 were highly active against Candida neoformans 32609 with MIC ≤ 0.125 µg/mL and showed broad-spectrum antifungal activity including against fluconazole-resistant Candida auris 891. In addition, compounds 6 and 22 demonstrated inhibitory effects on filamentation in the azole-resistant C. albicans isolate. Moreover, compounds 6 and 22 were minimally toxic to HUVECs and possessed weak inhibitory effects on the human CYP3A4 and CYP2D6. SARs and docking study further indicated that ortho-substituted groups in the terminal phenyl ring can promote the compounds to improve their antifungal activity.


Assuntos
Antifúngicos , Triazóis , Humanos , Antifúngicos/química , Triazóis/química , Testes de Sensibilidade Microbiana , Fluconazol/farmacologia , Candida albicans , Relação Estrutura-Atividade
5.
Int J Mol Sci ; 23(20)2022 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-36293157

RESUMO

Candida albicans is a typical opportunistic pathogen in humans that causes serious health risks in clinical fungal infections. The construction of mutant libraries has made remarkable developments in the study of C. albicans molecular and cellular biology with the ongoing advancements of gene editing, which include the application of CRISPR-Cas9 and novel high-efficient transposon. Large-scale genetic screens and genome-wide functional analysis accelerated the investigation of new genetic regulatory mechanisms associated with the pathogenicity and resistance to environmental stress in C. albicans. More importantly, sensitivity screening based on C. albicans mutant libraries is critical for the target identification of novel antifungal compounds, which leads to the discovery of Sec7p, Tfp1p, Gwt1p, Gln4p, and Erg11p. This review summarizes the main types of C. albicans mutant libraries and interprets their applications in morphogenesis, biofilm formation, fungus-host interactions, antifungal drug resistance, and target identification.


Assuntos
Antifúngicos , Candida albicans , Humanos , Antifúngicos/farmacologia , Farmacorresistência Fúngica/genética , Virulência , Genômica
6.
Acta Pharmacol Sin ; 42(5): 801-813, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-32796956

RESUMO

Grincamycins (GCNs) are a class of angucycline glycosides isolated from actinomycete Streptomyces strains that have potent antitumor activities, but their antitumor mechanisms remain unknown. In this study, we tried to identify the cellular target of grincamycin B (GCN B), one of most dominant and active secondary metabolites, using a combined strategy. We showed that GCN B-selective-induced apoptosis of human acute promyelocytic leukemia (APL) cell line NB4 through increase of ER stress and intracellular reactive oxygen species (ROS) accumulation. Using a strategy of combining phenotype, transcriptomics and protein microarray approaches, we identified that isocitrate dehydrogenase 1(IDH1) was the putative target of GCN B, and confirmed that GCNs were a subset of selective inhibitors targeting both wild-type and mutant IDH1 in vitro. It is well-known that IDH1 converts isocitrate to 2-oxoglutarate (2-OG), maintaining intracellular 2-OG homeostasis. IDH1 and its mutant as the target of GCN B were validated in NB4 cells and zebrafish model. Knockdown of IDH1 in NB4 cells caused the similar phenotype as GCN B treatment, and supplementation of N-acetylcysteine partially rescued the apoptosis caused by IDH1 interference in NB4 cells. In zebrafish model, GCN B effectively restored myeloid abnormality caused by overexpression of mutant IDH1(R132C). Taken together, we demonstrate that IDH1 is one of the antitumor targets of GCNs, suggesting wild-type IDH1 may be a potential target for hematological malignancies intervention in the future.


Assuntos
Antraquinonas/farmacologia , Antineoplásicos/farmacologia , Inibidores Enzimáticos/farmacologia , Glicosídeos/farmacologia , Isocitrato Desidrogenase/antagonistas & inibidores , Animais , Antraquinonas/metabolismo , Antineoplásicos/metabolismo , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Inibidores Enzimáticos/metabolismo , Glicosídeos/metabolismo , Humanos , Isocitrato Desidrogenase/genética , Isocitrato Desidrogenase/metabolismo , Ácidos Cetoglutáricos/metabolismo , Simulação de Acoplamento Molecular , Mutação , Ligação Proteica , Espécies Reativas de Oxigênio/metabolismo , Peixe-Zebra
7.
Bioorg Chem ; 101: 103982, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32534348

RESUMO

In order to develop novel antifungal agents, based on our previous work, a series of (2R,3R)-3-((3-substitutied-isoxazol-5-yl)methoxy)-2-(2,4-difluorophenyl)-1-(1H-1,2,4-triazol-1-yl) butan-2-ol (a1-a26) were designed and synthesized. All of the compounds exhibited good in vitro antifungal activities against eight human pathogenic fungi. Among them, compound a6 showed excellent inhibitory activity against Candida albicans and Candida parasilosis with MIC80 values of 0.0313 µg/mL. In addition, compounds a6, a9, a12, a13 and a14 exhibited moderate inhibitory activities against fluconazole-resistant isolates with MIC80 values ranging from 8 µg/mL to 16 µg/mL. Furthermore, compounds a6, a12 and a23 exhibited low inhibition profiles for CYP3A4. Clear SARs were analyzed, and the molecular docking experiment was carried out to further investigate the relationship between a6 and the target enzyme CYP51.


Assuntos
Antifúngicos/uso terapêutico , Candida albicans/efeitos dos fármacos , Isoxazóis/química , Simulação de Acoplamento Molecular/métodos , Triazóis/síntese química , Triazóis/uso terapêutico , Antifúngicos/farmacologia , Humanos , Estrutura Molecular , Relação Estrutura-Atividade , Triazóis/química
8.
Small ; 15(48): e1901550, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31115974

RESUMO

Carbon nanotubes (CNTs) as well-known nanomaterials are extensively studied and widely applied in various fields. Nitric acid (HNO3 ) is often used to treat CNTs for purification purposes and preparing oxidized CNTs for various applications. However, too little attention is paid to investigating the effect of HNO3 treatment on the optical properties of CNTs. In this work, it is observed for the first time that HNO3 -oxidized multiwalled carbon nanotubes (ox-MWCNTs) have strong electrochemiluminescence (ECL) activity, which enables ox-MWCNTs to become new and good ECL carbon nanomaterials after carbon quantum dots (CQDs) and graphene quantum dots (GQDs). Various characterization technologies, such as transmission electron microscope (TEM), X-ray photoelectron spectroscopy (XPS), and Raman spectroscopy, are used to reveal the relationship between ECL activity and surface states. The ECL behaviors of ox-MWCNTs are investigated in detail and a possible ECL mechanism is proposed. Finally, the new ECL nanomaterials of ox-MWCNTs are envisioned to have promising applications in sensitive ECL sensing and in the study of CNT-based catalysts.

9.
Protein Expr Purif ; 155: 21-26, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30414969

RESUMO

The interaction between TNF-α with TNFR1 triggers important signaling pathways inducing diverse cellular phenomena including inflammation, apoptosis, etc., and is involved in the pathogenesis and progression of numerous autoimmune diseases. The extracellular domain (ECD) of TNFR has been successfully used to clinically treat such TNF-associated diseases. However, large-scale production of these biological material via eukaryotic cell expression systems is usually costly owing to the culture medium and complicated growth conditions. This study aimed to extract pure soluble human TNFR1-ECD and investigate its biological activity, using a prokaryotic expression system. Recombinant vector pMCSG7-TNFR1-ECD was constructed via ligation-independent cloning. The His-tag fusion protein was expressed in E. coli and localized in inclusion bodies. Recombinant TNFR1-ECD was refolded and purified via nickel-affinity chromatography, tag cleavage, followed by cation-exchange chromatography or size-exclusion chromatography. A purity of over 95% and a yield of 9.3 mg protein per liter of bacterial culture media was obtained. The purified protein showed significant affinity of 2.15 nM towards human TNF-α and inhibited TNF-α-mediated cytotoxicity in L929 cells, with an ED50 of 0.10 µg/ml. It formed a self-associated oligomer with a KD of 1.15 µM, detected via microscale thermophoresis. We thus established a highly efficient approach to construct, express, and purify the recombinant protein of human TNFR1-ECD from a prokaryotic system. The antagonistic bioactivities in vitro indicate this protein as a prospective molecules for drug research against autoimmune diseases characterized by TNF-α overexpression.


Assuntos
Receptores Tipo I de Fatores de Necrose Tumoral/genética , Receptores Tipo I de Fatores de Necrose Tumoral/metabolismo , Linhagem Celular , Clonagem Molecular/métodos , Escherichia coli/genética , Fibroblastos/metabolismo , Humanos , Domínios Proteicos , Redobramento de Proteína , Receptores Tipo I de Fatores de Necrose Tumoral/química , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
10.
Molecules ; 21(5)2016 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-27164077

RESUMO

Total synthesis of naturally occurring spirobisnaphthalene palmarumycin CP17 and its methoxy analogues was first achieved through Friedel-Crafts acylation, Wolff-Kishner reduction, intramolecular cyclization, ketalization, benzylic oxidation, and demethylation using the inexpensive and readily available methoxybenzene, 1,2-dimethoxybenzene and 1,4-dimethoxybenzene and 1,8-dihydroxynaphthalene as raw materials. Demethylation with (CH3)3SiI at ambient temperature resulted in ring A aromatization and acetal cleavage to give rise to binaphthyl ethers. The antifungal activities of these spirobisnaphthalene derivatives were evaluated, and the results revealed that 5 and 9b exhibit EC50 values of 9.34 µg/mL and 12.35 µg/mL, respectively, against P. piricola.


Assuntos
Antifúngicos/síntese química , Naftalenos/síntese química , Compostos de Espiro/síntese química , Acilação , Anisóis/química , Antifúngicos/química , Antifúngicos/farmacologia , Ciclização , Fungos/efeitos dos fármacos , Estrutura Molecular , Naftalenos/química , Naftalenos/farmacologia , Compostos de Espiro/química , Compostos de Espiro/farmacologia
12.
Int J Womens Health ; 16: 1103-1111, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38895039

RESUMO

Objective: Additive human menopausal gonadotropin (HMG)/recombinant luteinizing hormone (r-LH) to follicle-stimulating hormone (FSH) can improve pregnancy outcomes in patients with poor ovarian response during assisted reproductive procedures. However, their effects on patients with normal ovarian response during such procedures are unclear, which formed the aim of this study. Methods: This retrospective study enrolled 456 infertile women who underwent in vitro fertilization or intracytoplasmic sperm injection treatment. Group 1 received FSH; Group 2 received FSH+HMG/r-LH; Group 3 received FSH+HMG+r-LH. Results: The age and Body Mass Index were significantly greater in Group III. The endometrial thickness was greater in Groups II and III, suggesting better endometrial receptivity. Better pregnancy and birth outcomes were seen in Group 3. In sub-cohorts of women older than 32 years old or with overweight/obesity, pregnancy and birth outcomes were also much better in Group 3, albeit without statistical significance. Conclusion: The addition of both HMG and r-LH to FSH may improve the chance of infertile women with normal ovarian responses to have more success in having live birth babies, specifically in those over 32 years of age or with overweight/obese patients who typically face challenges in conceiving and sustaining a pregnancy.

13.
J Med Chem ; 67(8): 6238-6252, 2024 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-38598688

RESUMO

Thirty-one novel albaconazole derivatives were designed and synthesized based on our previous work. All compounds exhibited potent in vitro antifungal activities against seven pathogenic fungi. Among them, tetrazole compound D2 was the most potent antifungal with MIC values of <0.008, <0.008, and 2 µg/mL against Candida albicans, Cryptococcus neoformans, and Aspergillus fumigatus, respectively, the three most common and critical priority pathogenic fungi. In addition, compound D2 also exhibited potent activity against fluconazole-resistant C. auris isolates. Notably, compound D2 showed a lower inhibitory activity in vitro against human CYP450 enzymes as well as a lower inhibitory effect on the hERG K+ channel, indicating a low risk of drug-drug interactions and QT prolongation. Moreover, with improved pharmacokinetic profiles, compound D2 showed better in vivo efficacy than albaconazole at reducing fungal burden and extending the survival of C. albicans-infected mice. Taken together, compound D2 will be further investigated as a promising candidate.


Assuntos
Antifúngicos , Candida albicans , Cryptococcus neoformans , Testes de Sensibilidade Microbiana , Tetrazóis , Antifúngicos/farmacologia , Antifúngicos/síntese química , Antifúngicos/química , Antifúngicos/uso terapêutico , Tetrazóis/farmacologia , Tetrazóis/química , Tetrazóis/síntese química , Tetrazóis/farmacocinética , Tetrazóis/uso terapêutico , Animais , Humanos , Candida albicans/efeitos dos fármacos , Camundongos , Cryptococcus neoformans/efeitos dos fármacos , Relação Estrutura-Atividade , Aspergillus fumigatus/efeitos dos fármacos , Descoberta de Drogas , Farmacorresistência Fúngica/efeitos dos fármacos , Candidíase/tratamento farmacológico , Inibidores das Enzimas do Citocromo P-450/farmacologia , Inibidores das Enzimas do Citocromo P-450/síntese química , Inibidores das Enzimas do Citocromo P-450/química , Sistema Enzimático do Citocromo P-450/metabolismo
14.
ACS Omega ; 9(11): 12478-12499, 2024 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-38524433

RESUMO

In the respiratory chain of the majority of aerobic organisms, the enzyme alternative oxidase (AOX) functions as the terminal oxidase and has important roles in maintaining metabolic and signaling homeostasis in mitochondria. AOX endows the respiratory system with flexibility in the coupling among the carbon metabolism pathway, electron transport chain (ETC) activity, and ATP turnover. AOX allows electrons to bypass the main cytochrome pathway to restrict the generation of reactive oxygen species (ROS). The inhibition of AOX leads to oxidative damage and contributes to the loss of adaptability and viability in some pathogenic organisms. Although AOXs have recently been identified in several organisms, crystal structures and major functions still need to be explored. Recent work on the trypanosome alternative oxidase has provided a crystal structure of an AOX protein, which contributes to the structure-activity relationship of the inhibitors of AOX. Here, we review the current knowledge on the development, structure, and properties of AOXs, as well as their roles and mechanisms in plants, animals, algae, protists, fungi, and bacteria, with a special emphasis on the development of AOX inhibitors, which will improve the understanding of respiratory regulation in many organisms and provide references for subsequent studies of AOX-targeted inhibitors.

15.
Eur J Med Chem ; 275: 116637, 2024 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-38959728

RESUMO

Life-threatening invasive fungal infections pose a serious threat to human health. A series of novel triazole derivatives bearing a pyrazole-methoxyl moiety were designed and synthesized in an effort to obtain antifungals with potent, broad-spectrum activity that are less susceptible to resistance. Most of these compounds exhibited moderate to excellent in vitro antifungal activities against Candida albicans SC5314 and 10,231, Cryptococcus neoformans 32,609, Candida glabrata 537 and Candida parapsilosis 22,019 with minimum inhibitory concentration (MIC) values of ≤0.125 µg/mL to 0.5 µg/mL. Use of recombinant Saccharomyces cerevisiae strains showed compounds 7 and 10 overcame the overexpression and resistant-related mutations in ERG11 of S. cerevisae and several pathogenic Candida spp. Despite being substrates of the C. albicans and Candida auris Cdr1 drug efflux pumps, compounds 7 and 10 showed moderate potency against five fluconazole (FCZ)-resistant fungi with MIC values from 2.0 µg/mL to 16.0 µg/mL. Growth kinetics confirmed compounds 7 and 10 had much stronger fungistatic activity than FCZ. For C. albicans, compounds 7 and 10 inhibited the yeast-to-hyphae transition, biofilm formation and destroyed mature biofilm more effectively than FCZ. Preliminary mechanism of action studies showed compounds 7 and 10 blocked the ergosterol biosynthesis pathway at Erg11, ultimately leading to cell membrane disruption. Further investigation of these novel triazole derivatives is also warranted by their predicted ADMET properties and low cytotoxicity.


Assuntos
Antifúngicos , Candida , Testes de Sensibilidade Microbiana , Pirazóis , Triazóis , Antifúngicos/farmacologia , Antifúngicos/síntese química , Antifúngicos/química , Triazóis/química , Triazóis/farmacologia , Triazóis/síntese química , Pirazóis/química , Pirazóis/farmacologia , Pirazóis/síntese química , Relação Estrutura-Atividade , Candida/efeitos dos fármacos , Estrutura Molecular , Relação Dose-Resposta a Droga , Cryptococcus neoformans/efeitos dos fármacos , Humanos , Farmacorresistência Fúngica/efeitos dos fármacos , Saccharomyces cerevisiae/efeitos dos fármacos , Candida albicans/efeitos dos fármacos
16.
J Med Chem ; 67(5): 4007-4025, 2024 Mar 14.
Artigo em Inglês | MEDLINE | ID: mdl-38381075

RESUMO

Invasive fungal infections pose a serious threat to public health and are associated with high mortality and incidence rates. The development of novel antifungal agents is urgently needed. Based on hit-to-lead optimization, a series of 2,4,6-trisubstituted triazine hydrazone compounds were designed, synthesized, and biological evaluation was performed, leading to the identification of compound 28 with excellent in vitro synergy (FICI range: 0.094-0.38) and improved monotherapy potency against fluconazole-resistant Candida albicans and Candida auris (MIC range: 1.0-16.0 µg/mL). Moreover, 28 exhibited broad-spectrum antifungal activity against multiple pathogenic strains. Furthermore, 28 could inhibit hyphal and biofilm formation, which may be related to its ability to disrupt the fungal cell wall. Additionally, 28 significantly reduced the CFU in a mouse model of disseminated infection with candidiasis at a dose of 10 mg/kg. Overall, the triazine-based hydrazone compound 28 with low cytotoxicity, hemolysis, and favorable ADME/T characteristics represents a promising lead to further investigation.


Assuntos
Antifúngicos , Candidíase , Animais , Camundongos , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Testes de Sensibilidade Microbiana , Fluconazol/farmacologia , Candida albicans , Candidíase/tratamento farmacológico , Candidíase/microbiologia
17.
Nat Commun ; 15(1): 4131, 2024 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-38755250

RESUMO

The transition between yeast and hyphae is crucial for regulating the commensalism and pathogenicity in Candida albicans. The mechanisms that affect the invasion of hyphae in solid media, whose deficiency is more related to the pathogenicity of C. albicans, have not been elucidated. Here, we found that the disruption of VAM6 or VPS41 which are components of the homotypic vacuolar fusion and protein sorting (HOPS) complex, or the Rab GTPase YPT72, all responsible for vacuole fusion, led to defects in hyphal growth in both liquid and solid media, but more pronounced on solid agar. The phenotypes of vac8Δ/Δ and GTR1OE-vam6Δ/Δ mutants indicated that these deficiencies are mainly caused by the reduced mechanical forces that drive agar and organs penetration, and confirmed that large vacuoles are required for hyphal mechanical penetration. In summary, our study revealed that large vacuoles generated by vacuolar fusion support hyphal penetration and provided a perspective to refocus attention on the role of solid agar in evaluating C. albicans invasion.


Assuntos
Candida albicans , Proteínas Fúngicas , Hifas , Vacúolos , Candida albicans/metabolismo , Candida albicans/genética , Hifas/metabolismo , Hifas/crescimento & desenvolvimento , Hifas/genética , Vacúolos/metabolismo , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/genética , Animais , Camundongos , Proteínas rab de Ligação ao GTP/metabolismo , Proteínas rab de Ligação ao GTP/genética , Candidíase/microbiologia , Proteínas de Transporte Vesicular/metabolismo , Proteínas de Transporte Vesicular/genética , Feminino , Fusão de Membrana
18.
Zhonghua Yan Ke Za Zhi ; 49(2): 163-9, 2013 Feb.
Artigo em Zh | MEDLINE | ID: mdl-23714035

RESUMO

OBJECTIVE: To investigate the influence of TGF-ß2 antisense oligonucleotide (ASON) on preventing corneal scar hyperplasia in rabbits and to provide experimental evidence for its clinical application. METHODS: It was an experimental study. One hundred and ninety two New Zealand white rabbits were randomly divided into 4 groups (groups A, B, C and D). Corneal injury models were established in all groups. There were 48 experimental animals in each group. TGF-ß2 ASON was dropped into right eyes in group A, dexamethasone was dropped into right eyes in group B, deionized water was dropped into right eyes in group C and nothing was dropped into right eyes in group D after the operation. The corneas were surgically removed and assessed by hematoxylin eosin (HE) staining, immunohistochemical study and real time PCR at four different time points (4 d, 7 d, 14 d and 28 d) after surgery. RESULTS: HE staining: at the same time point, fibroblasts in the groups A and B were significantly fewer than that in the groups C and D, the difference was statistically significant (P < 0.05), but there was no significant difference between groups A and B or groups C and D. Immunohistochemical observation found that the expression of α-smooth muscle actin (α-SMA) positive fibroblasts could be observed by the 4th day (9.44 ± 0.47/HP), reached a climax by the 7th day (12.50 ± 0.81/HP), and returned to the baseline levels by the 14th day (0.85 ± 0.43/HP) in the group A, which was similar to that in the group B (9.49 ± 0.95, 12.42 ± 0.70, 0.86 ± 0.79/HP) at the same time point (P > 0.05), but it was significantly fewer than that in the group C(20.14 ± 0.78, 18.19 ± 1.28, 4.87 ± 0.58/HP) and group D(20.21 ± 0.92, 18.25 ± 1.39, 5.00 ± 2.217/HP), which was statistical significant (P < 0.05). The staining intensity of fibronectin (FN) in groups A and B was significantly weaker than that in groups C and D. Real time PCR analysis showed that at each time point, the expression of TGF-ß2 mRNAs in groups A and B was significantly lower than that in groups C and D (P < 0.05). CONCLUSIONS: TGF-ß2 ASON can effectively prevent the proliferation of corneal tissue by inhibiting the activity of TGF-ß2 after injury. The early stage of corneal repair is 7 days after injury, so it is important to use TGF-ß2 ASON at this stage to inhibit the scar hyperplasia. In addition, it is safe to apply TGF-ß2 ASON topically to protect the cornea from obvious side effects.


Assuntos
Cicatriz/patologia , Córnea/efeitos dos fármacos , Córnea/patologia , Doenças da Córnea/patologia , Fator de Crescimento Transformador beta2/farmacologia , Animais , Feminino , Hiperplasia/prevenção & controle , Masculino , Oligonucleotídeos Antissenso , Coelhos , Fator de Crescimento Transformador beta2/genética , Cicatrização
19.
Zhonghua Yan Ke Za Zhi ; 49(10): 927-33, 2013 Oct.
Artigo em Zh | MEDLINE | ID: mdl-24433696

RESUMO

OBJECTIVE: To investigate the preventive and therapeutic effects of agent calcium dobesilate(CDO) with different doses on the galactose cataract of rats. METHODS: We chose fifty Wistar rats at 20- day old. Then, they were divided into 3 groups at random. Choose 10 rats as the control group and gave normal diet; 10 rats as the model group and fed with Gal solution ( drink 12.5% Gal solution from 1 to 7 days and 10%Gal solution from 8 to 21 days except for normal diet ) ; 30 rats as the treatment group and fed with the same Gal solution as the model group, besides they were divided into high dosage group, medium dosage group and low dosage group equally and gave 300 mg×kg(-1)×d(-1), 150 mg×kg(-1)×d(-1), 75 mg×kg(-1)×d(-1) dose of calcium dobesilate respectively from the first day to the end of experiment. The experiment lasts 21 days. Lens opacity were observed and recorded by slit-lamp examination regularly. Activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) and content of malondialdehyde (MDA) were determined to estimate the effect of CDO . Lens fibers changes and Histological changes were observed by scanning electron microscope (SEM) and light microscope (LM) separately. The apoptosis rate of lens epithelium were determined by TUNEL assay. RESULTS: The appearance of Lens opacity in model group was more quickly than that in treatment group in model group, 3 eyes in degree IV, 7 eyes in degree V, while in treatment group, 5 eyes in degree III, 3 eyes in degree IV, 2 eyes in degree V (H = 7.12, P < 0.05). The activity of SOD and GSH-px in treatment group is higher than mode group, but lower than control group on 8th day, there was difference noticed in the activity of SOD (50.01 ± 1.19), (39.39 ± 1.70) , treatment group (46.57 ± 1.09, 46.42 ± 0.87, 45.70 ± 1.46) U/mgProt (F = 88.70, P < 0.05) and the activity of GSH-px (42.92 ± 0.97) , (12.70 ± 1.17) , treatment group (29.16 ± 1.05, 29.08 ± 0.98, 28.25 ± 0.98) nmol/mgprot (F = 1071.89, P < 0.05) ]in 3 groups. The content of MDA in model group (3.43 ± 0.15)nmol/mgprot is higher than treatment group (2.89 ± 0.11, 2.99 ± 0.12, 2.99 ± 0.09)nmol/mgprot (F = 64.62; P < 0.05). There were no statistic significant differences among high dosage group, medium dosage group and low dosage group . The texture of lens fibres detected by SEM in the rats of model was more disorder than treatment group. After HE staining, Lens epithelial cell detected by LM in control group have a clear structure, however, Lens epithelial cell both in model group and treatment group have changed from the initial single layer to multi-storey. Junction between lenses fibers became decreased even disappeared . The apoptosis rate of lens epithelium in treatment group[(2.37 ± 0.17)%, (2.46 ± 0.26)%, (2.79 ± 0.41)%] is higher than control group (0.23 ± 0.07) %, but is much fewer than model group (4.99 ± 0.51) % (χ(2) = 40.41;P < 0.05). CONCLUSION: CDO with different doses could protect lens of rats against galactose damage and there were no significant differences among the different doses of groups.


Assuntos
Dobesilato de Cálcio/farmacologia , Catarata/prevenção & controle , Animais , Dobesilato de Cálcio/uso terapêutico , Catarata/induzido quimicamente , Catarata/tratamento farmacológico , Galactose , Ratos , Ratos Wistar
20.
Zhonghua Yan Ke Za Zhi ; 49(12): 1118-23, 2013 Dec.
Artigo em Zh | MEDLINE | ID: mdl-24499699

RESUMO

OBJECTIVE: To observe the visual quality of aspherical intraocular lens (IOL) implantation in traumatic cataract patients. METHODS: Prospective clinical study.96 traumatic cataract patients (96 eyes) suffered from penetrating corneal trauma chosen from the first affiliated hospital of Zhengzhou university during June 2009 to June 2012. They were divided into two groups based on the different type of intraocular lens. The experimental group (48 eyes) was implanted with aspherical IOL and the control group (48 eyes) was implanted with traditional sphere IOL.Uncorrected visual acuity (UCVA), best corrected visual acuity (BCVA), contrast sensitivity (CS) and stereoscopic vision were observed at 1 month, 3 months, and 6 months after surgery. At the same time, questionnaire survey about the satisfaction of patients was also performed. The t test was used to compare the preoperative general condition, postoperative visual acuity, contrast sensitivity and stereoscopic vision of the two groups, and the rank sum test was used to compare the astigmatism and the satisfaction of patients. RESULTS: There was no significant difference in UCVA (t = 1.37, 1.28,0.71, P > 0.05) between the experimental group (0.56 ± 0.22, 0.68 ± 0.13,0.84 ± 0.15) and the control group (0.51 ± 0.17, 0.61 ± 0.20,0.81 ± 0.17) at three time points. There was no significant difference in BCVA (t = 0.87, 1.38, 1.39, P > 0.05) between the experimental group (0.62 ± 0.13, 0.74 ± 0.21, 0.87 ± 0.10) and the control group (0.57 ± 0.25,0.69 ± 0.22,0.84 ± 0.15) . The same result happened in stereoscopic vision at 6 months after surgery (far stereopsis:123.5 ± 7.8 vs 126. 9 ± 5.9, t = 0.64, P > 0.05;near stereopsis:90.5 ± 7.8 vs 95.2 ± 3.5; t = 1.36, P > 0.05) between experimental group and control group. The contrast sensitivity of the experimental group in every stage (3 c/d:1.52 ± 0.18, 6 c/d:1.68 ± 0.19, 12 c/d:1.29 ± 0.14, 18 c/d:1.04 ± 0.20) was superior to the control group (3 c/d:1.49 ± 0.27, 6 c/d:1.57 ± 0.21, 12 c/d:1.14 ± 0.20, 18 c/d:0.85 ± 0.14) , especially on the glare sensitivity (the experimental group:3 c/d:1.40 ± 0.15, 6 c/d:1.52 ± 0.22, 12 c/d:1.21 ± 0.18, 18 c/d:0.91 ± 0.14, the control group:3 c/d:1.13 ± 0.13, 6 c/d:1.13 ± 0.28, 12 c/d:0.92 ± 0.13, 18 c/d:0.54 ± 0.16) Compared two groups of difference have statistical significance (free from glare:3 c/d:t = 2.829, 6 c/d:t = 4.092, 12 c/d:t = 3.055, 18 c/d:t = 2.093;glare:3 c/d:t = 2.650, 6 c/d:t = 3.105, 12 c/d:t = 3.395, 18 c/d:t = 2.215;P < 0.05) .Questionnaire survey showed the experimental group (72.9%) was statistically significantly higher (t = 3.016, P < 0.05) than that in the control group (54.1%) on the satisfaction of patients. CONCLUSIONS: The visual quality with implantation of aspherical IOL in traumatic cataract patients is superior to traditional sphere IOL. Aspherical IOL is more appropriate for the patients with small and peripheral corneal scar.It can reduce the visual function damage to minimum caused by trauma.


Assuntos
Catarata/terapia , Implante de Lente Intraocular , Acuidade Visual , Adolescente , Adulto , Catarata/etiologia , Criança , Traumatismos Oculares/complicações , Humanos , Pessoa de Meia-Idade , Estudos Prospectivos , Resultado do Tratamento , Adulto Jovem
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