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1.
BMC Neurosci ; 21(1): 13, 2020 03 30.
Artigo em Inglês | MEDLINE | ID: mdl-32228575

RESUMO

BACKGROUND: Patients positive for anti-glutamic acid decarboxylase 65 (GAD65) antibodies have attracted increasing attention. Their clinical manifestations are highly heterogeneous and can be comorbid with tumors. Currently, there is no consensus on the therapeutic regimen for anti-GAD65-associated neurological diseases due to the clinical complexity, rarity and sporadic distribution. We reported six anti-GAD65 autoimmune encephalitis (AE) patients who received intravenous methylprednisolone (IVMP) or immunoglobulin (IVIG) or both. Then, we evaluated the therapeutic effect of both by summarizing results in previous anti-GAD65 AE patients from 70 published references. RESULTS: Our six patients all achieved clinical improvements in the short term. Unfortunately, there was no significant difference between IVMP and IVIG in terms of therapeutic response according to the previous references, and the effectiveness of IVMP and IVIG was 45.56% and 36.71%, respectively. We further divided the patients into different subgroups according to their prominent clinical manifestations. The response rates of IVMP and IVIG were 42.65% and 32.69%, respectively, in epilepsy patients; 60.00% and 77.78%, respectively, in patients with stiff-person syndrome; and 28.57% and 55.56%, respectively, in cerebellar ataxia patients. Among 29 anti-GAD65 AE patients with tumors, the response rates of IVMP and IVIG were 29.41% and 42.11%, respectively. There was no significant difference in effectiveness between the two regimens among the different subgroups. CONCLUSION: Except for stiff-person syndrome, we found that this kind of AE generally has a poor response to IVMP or IVIG. Larger prospective studies enrolling large numbers of patients are required to identify the optimal therapeutic strategy in the future.


Assuntos
Encefalite/tratamento farmacológico , Glucocorticoides/administração & dosagem , Glutamato Descarboxilase/imunologia , Doença de Hashimoto/tratamento farmacológico , Imunoglobulinas/administração & dosagem , Fatores Imunológicos/administração & dosagem , Metilprednisolona/administração & dosagem , Administração Intravenosa , Adulto , Autoanticorpos/sangue , Autoanticorpos/líquido cefalorraquidiano , Encéfalo/diagnóstico por imagem , Encéfalo/patologia , Encefalite/diagnóstico por imagem , Encefalite/imunologia , Feminino , Doença de Hashimoto/diagnóstico por imagem , Doença de Hashimoto/imunologia , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Resultado do Tratamento
2.
Blood ; 132(11): 1146-1158, 2018 09 13.
Artigo em Inglês | MEDLINE | ID: mdl-30054295

RESUMO

Mature T-cell lymphomas, including peripheral T-cell lymphoma (PTCL) and extranodal NK/T-cell lymphoma (NKTL), represent a heterogeneous group of non-Hodgkin lymphomas with dismal outcomes and limited treatment options. To determine the extent of involvement of the JAK/STAT pathway in this malignancy, we performed targeted capture sequencing of 188 genes in this pathway in 171 PTCL and NKTL cases. A total of 272 nonsynonymous somatic mutations in 101 genes were identified in 73% of the samples, including 258 single-nucleotide variants and 14 insertions or deletions. Recurrent mutations were most frequently located in STAT3 and TP53 (15%), followed by JAK3 and JAK1 (6%) and SOCS1 (4%). A high prevalence of STAT3 mutation (21%) was observed specifically in NKTL. Novel STAT3 mutations (p.D427H, E616G, p.E616K, and p.E696K) were shown to increase STAT3 phosphorylation and transcriptional activity of STAT3 in the absence of cytokine, in which p.E616K induced programmed cell death-ligand 1 (PD-L1) expression by robust binding of activated STAT3 to the PD-L1 gene promoter. Consistent with these findings, PD-L1 was overexpressed in NKTL cell lines harboring hotspot STAT3 mutations, and similar findings were observed by the overexpression of p.E616K and p.E616G in the STAT3 wild-type NKTL cell line. Conversely, STAT3 silencing and inhibition decreased PD-L1 expression in STAT3 mutant NKTL cell lines. In NKTL tumors, STAT3 activation correlated significantly with PD-L1 expression. We demonstrated that STAT3 activation confers high PD-L1 expression, which may promote tumor immune evasion. The combination of PD-1/PD-L1 antibodies and STAT3 inhibitors might be a promising therapeutic approach for NKTL, and possibly PTCL.


Assuntos
Antígeno B7-H1/biossíntese , Regulação Neoplásica da Expressão Gênica , Mutação de Sentido Incorreto , Proteínas de Neoplasias/biossíntese , Fator de Transcrição STAT3/biossíntese , Transdução de Sinais , Substituição de Aminoácidos , Antígeno B7-H1/antagonistas & inibidores , Antígeno B7-H1/genética , Linhagem Celular Tumoral , Humanos , Linfoma Extranodal de Células T-NK , Proteínas de Neoplasias/genética , Fator de Transcrição STAT3/genética
3.
Mikrochim Acta ; 186(8): 594, 2019 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-31372831

RESUMO

A method is described for the colorimetric determination of the activity of CpG methyltransferase (M.SssI). It is based on (a) the crosslinking effect between dsDNA-modified gold nanoparticles (AuNPs) and graphene oxide (GO), and (b) an amplification reaction with the aid of a nicking enzyme. To avoid the aggregation of AuNPs (which would produce false signals), a hairpin DNA was connected to the AuNPs. Thus, the red color of the solution (measured at 530 nm) increases linearly with the activity of M.SssI from 0.2 to 60 U·mL-1, and the limit of detection is 67 U·mL-1. This is superior to some reported strategies. The method was successfully applied to analyze spiked serum samples. Conceivably, it represents a powerful tool for use in drug development and diagnosis. Graphical abstracts A method based on the conjugated cross-linking effect between dsDNA modified Au NPs and GO coupled with an amplification reaction of nicking enzyme has been developed for colorimetric detection of the activity of CpG methyltransferase (M.SssI).

4.
Mikrochim Acta ; 186(11): 732, 2019 10 30.
Artigo em Inglês | MEDLINE | ID: mdl-31667627

RESUMO

The authors describe an electrochemical and an optical method for the determination of As(V) by using iron oxyhydroxide (FeOOH) nanorods that display peroxidase-mimicking activity. The nanorods catalyze the oxidation of substrate ABTS by H2O2 to form a green product with an absorption maximum at 418 nm. If, however, As(V) is electrostatically adsorbed on the nanorods, the oxidation is gradually inhibited. A colorimetric assay was worked out based on these findings. Response is linear in the 0 to 8 ppb and 8 to 200 ppb As(V) concentration range, and the detection limit is 0.1 ppb. Even higher sensitivity is achieved in an electrochemical method which is based on the excellent electrical conductivity of FeOOH nanorods. Electrochemical analysis of As(V) was achieved by first adsorbing As(V) on the nanorods. This inhibits the ABTS reduction current signal, best measured at a potential of 150 mV (vs. Ag/AgCl). The linear range extends from 0.04 to 200 ppb, and the detection limit is as low as 12 ppt. Graphical abstract Schematic representation of FeOOH nanorod-based colorimetric and electrochemical assays for arsenate (As(V)). As(V) adsorbed on FeOOH nanorods inhibits the peroxidase-mimicking activity of nanorods, and a colorimetric and electrochemical dual-signal assay was constructed to achieve sensitive determination of As(V).


Assuntos
Arseniatos/análise , Colorimetria/métodos , Técnicas Eletroquímicas/métodos , Compostos Férricos/química , Nanotubos/química , Poluentes Químicos da Água/análise , Benzotiazóis/química , Materiais Biomiméticos/química , Catálise , Água Potável/análise , Peróxido de Hidrogênio/química , Lagos/análise , Limite de Detecção , Peroxidase/química , Rios/química , Ácidos Sulfônicos/química
5.
Mikrochim Acta ; 186(4): 241, 2019 03 13.
Artigo em Inglês | MEDLINE | ID: mdl-30868262

RESUMO

The activity of terminal deoxynucleotidyl transferase (TdTase) is a biomarker for routine diagnosis of acute leukemia. A method has been developed for the determination of TdTase activity. It is based on the use of silver nanoclusters (AgNCs) whose yellow fluorescence is enhanced by an in-situ grown DNA tail of TdTase-polymerized and guanine-rich DNA at the 3' end of a hairpin DNA. The fluorescence, best measured at excitation/emission peaks of 530/585 nm, increases linearly in the 1 to 35 mU mL-1 TdTase activity range. The detection limit is 0.8 mU mL-1. The method is cost-efficient, selective and convenient. It integrates enhancement of the fluorescence of AgNCs and target recognition into a single process. Graphical abstract Schematic presentation of a method for determination of TdTase activity. It is based on AgNCs fluorescence enhanced by in-situ grown TdTase-polymerized G-rich DNA tail. The method integrates AgNCs fluorescence enhancement and the target recognition into a single process.


Assuntos
DNA Nucleotidilexotransferase/sangue , DNA/química , Ensaios Enzimáticos/métodos , Nanopartículas Metálicas/química , Sequência de Bases , Biomarcadores/sangue , Técnicas Biossensoriais/métodos , DNA/genética , Fluorescência , Humanos , Sequências Repetidas Invertidas , Leucemia/diagnóstico , Limite de Detecção , Prata/química , Espectrometria de Fluorescência/métodos
6.
Cell Tissue Res ; 374(3): 653-666, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30073544

RESUMO

Testicular dysfunction is one of the serious secondary complications in diabetes. Lycium barbarum polysaccharide (LBP) has long been considered to possess a wide range of beneficial properties including antiaging, anticancer and reproductive-enhancing. Abnormal autophagy was reported to play a significant role in accelerating diabetic reproductive injury. However, the autophagy regulation mechanism of LBP on diabetic testicular dysfunction is incompletely understood. We investigate the protective effects of LBP on diabetic testicular dysfunction and its underlying mechanism with different approaches. Protective effects of LBP (40 mg/kg) on testicular functions were assessed through the use of sperm parameters, testosterone levels and hematoxylin and eosin staining. Antioxidant capacity and serum malondialdehyde levels were determined using assay kits. Immune intensity of Beclin-1 and LC3I in testes was detected by immunofluorescence staining. Western blot analysis was used to detect expressions of p-PI3K, Akt, p-Akt, Beclin-1, LC3I and LC3II proteins. Q-PCR was used to evaluate Beclin-1 and LC3I mRNA expressions in testis. Administration of LBP (40 mg/kg) considerably recovered testicular function, obviously improved testicular histopathologic structure and significantly increased antioxidant enzyme activities. Immunofluorescence staining showed that immune intensity of Beclin-1 and LC3I significantly decreased in the LBP 40 mg/kg group. The results of Q-PCR and western blot analysis showed that LBP 40 mg/kg significantly downregulated Beclin-1 and LC3I protein expressions upregulated p-PI3K and p-Akt protein expressions and decreased Beclin-1 and LC3I mRNA expressions compared with diabetic mice. In conclusion, inhibition of PI3K/Akt pathway-mediated testicular excessive autophagy may be a target for protective effects of LBP on diabetic testicular dysfunction.


Assuntos
Autofagia , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/fisiopatologia , Medicamentos de Ervas Chinesas/uso terapêutico , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Testículo/patologia , Testículo/fisiopatologia , Animais , Autofagossomos/efeitos dos fármacos , Autofagossomos/metabolismo , Autofagossomos/ultraestrutura , Autofagia/efeitos dos fármacos , Autofagia/genética , Proteínas Relacionadas à Autofagia/genética , Proteínas Relacionadas à Autofagia/metabolismo , Proteína Beclina-1/metabolismo , Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Experimental/genética , Medicamentos de Ervas Chinesas/farmacologia , Masculino , Camundongos Endogâmicos ICR , Proteínas Associadas aos Microtúbulos/metabolismo , Estresse Oxidativo/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Espermatozoides/metabolismo , Testículo/efeitos dos fármacos , Testosterona/sangue
7.
Mikrochim Acta ; 185(5): 280, 2018 05 03.
Artigo em Inglês | MEDLINE | ID: mdl-29725866

RESUMO

A method is reported for the fluorometric quantitation of microRNA. It is making use of a luminescent probe deribed from terbium(III) ion whose fluorescence is sensitized with a guanine-rich (G-rich) nucleotide. The probe has a large Stokes' shift and strong and sharp emission bands. The assay relies on the wide substrate specificity of terminal deoxynucleotidyl transferase (TdTase), which catalyzes the formation of long G-rich nucleotides when using microRNA primer as a trigger to start the polymerization. The addition of Tb(III) induces the formation of a G-quadruplex from the G-rich nucleotide, and this strongly enhances the green fluorescence of Tb(III) (peaking at 545 nm upon photoexcitation at 290 nm). Specifically, microRNA-21 was chosen as the analyte. The fluorescence intensity of Tb(III) increases linearly in the 1 pM to 1 nM microRNA concentration range, and the detection limit is as low as 0.11 pM. The method can distinguish between family members of microRNA and performs excellently even when applied to extracts of cancer cells. Graphical abstract A fluorometric technique is reported for the determination of microRNA. It is based on signal enhancement based on the sensitization of terbium(III) via a guanine-rich nucleotide sequence. Klenow Fragment exo- (KFexo-) generates DNA sequence at the 3'-OH of microRNA, and terminal deoxynucleotidyl transferase (TdTase) catalyzes the formation of long G-rich nucleotides.


Assuntos
Técnicas Biossensoriais/métodos , DNA Nucleotidilexotransferase/metabolismo , Nucleotídeos de Guanina/química , Nucleotídeos de Guanina/metabolismo , Medições Luminescentes/métodos , MicroRNAs/análise , Térbio/química , Células A549 , Humanos , Células MCF-7
8.
Heliyon ; 9(12): e22605, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38107270

RESUMO

Hepatocellular carcinoma (HCC) is a common malignant tumor of the digestive system with a low early diagnosis rate. Owing to the side effects, tolerance, and patient contraindications of existing therapies, effective drug treatments for HCC remain a major clinical challenge. However, using approved or investigational drugs not initially intended for cancer therapy is a promising strategy for resolving this problem because their safety have been tested in clinic. Therefore, this study evaluated differentially expressed genes between liver cancer and normal tissues in a cohort of patients with HCC from The Cancer Genome Atlas and applied them to query a connectivity map to identify candidate anti-HCC drugs. As a result, fluphenazine was identified as a candidate for anti-HCC therapy in vitro and in vivo. Fluphenazine suppressed HCC cell proliferation and migration and induced cell cycle arrest and apoptosis, possibly owing to disrupted lysosomal function, blocking autophagy flux. Additionally, in vivo studies demonstrated that fluphenazine suppresses HCC subcutaneous xenografts growth without causing severe side effects. Strikingly, fluphenazine could be used as an analgesic to alleviate oxaliplatin-induced pain as well as pain related anxiety-like behavior. Therefore, fluphenazine could be a novel liver cancer treatment candidate.

9.
Otolaryngol Head Neck Surg ; 169(5): 1247-1258, 2023 11.
Artigo em Inglês | MEDLINE | ID: mdl-37264983

RESUMO

OBJECTIVE: To investigate the role of H+ /K+ ATPase in the proliferation of pepsin-induced vocal cord leukoplakia (VCL) cells. STUDY DESIGN: Translation research. SETTING: Affiliated Hospital of University. METHODS: Immunohistochemistry was used to detect pepsin, H+ /K+ ATPase (ATP4A and ATP4B subunits) in VCL cells with varying degrees of dysplasia. After primary cultures of VCL cells had been established, the effects of acidified pepsin on the proliferation, autophagy, and H+ /K+ -ATPase distribution of VCL cells were investigated. RESULTS: The levels of pepsin, ATP4A, and ATP4B were significantly higher in VCL tissue with moderate-to-severe dysplasia than in normal tissue (p < .05); these levels gradually increased according to dysplasia severity. The expression levels of ATP4A and ATP4B were significantly correlated with the amount of pepsin in VCL cells (p < .01). Acidified pepsin enhanced the levels of proliferation and autophagy in human VCL epithelial cells. The cloning- and autophagy-promoting effects of acidified pepsin on VCL cells were partially reversed by pantoprazole; these effects were completely blocked by the autophagy inhibitor chloroquine. Finally, acidified pepsin promoted the colocalization of H+ /K+ -ATPase and lysosomes in VCL cells; it also mediated lysosome acidification. CONCLUSION: Pepsin and H+ /K+ -ATPase may contribute to the progression of VCL. Specifically, acidified pepsin may regulate lysosome acidification by promoting lysosomal localization of H+ /K+ -ATPase.


Assuntos
Doenças da Laringe , Pepsina A , Humanos , Prega Vocal/metabolismo , Autofagia , Células Epiteliais/metabolismo , Adenosina Trifosfatases , Proliferação de Células , Leucoplasia/metabolismo
10.
Pharmacol Ther ; 240: 108229, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-35700828

RESUMO

Cyclin-dependent kinase (CDK) 7 is best characterized for the ability to regulate biological processes, including the cell cycle and gene transcription. Abnormal CDK7 activity is observed in various tumours and represents a driving force for tumourigenesis. Therefore, CDK7 may be an appealing target for cancer treatment. Whereas, the enthusiasm for CDK7-targeted therapeutic strategy is mitigated due to the widely possessed belief that this protein is essential for normal cells. Indeed, the fact confronts the consensus. This is the first review to introduce the role of CDK7 in pan-cancers via a combined analysis of comprehensive gene information and (pre)clinical research results. We also discuss the recent advances in protein structure and summarize the understanding of mechanisms underlying CDK7 function. These endeavours highlight the pivotal roles of CDK7 in tumours and may contribute to the development of effective CDK7 inhibitors within the strategy of structure-based drug discovery for cancer therapy.


Assuntos
Neoplasias , Humanos , Ciclo Celular , Neoplasias/metabolismo , Descoberta de Drogas
11.
Ann Clin Transl Neurol ; 9(8): 1116-1124, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35699312

RESUMO

OBJECTIVE: Fear aura has traditionally been considered relevant to epileptic discharges from mesial temporal areas, and few studies have investigated its effect on surgical outcome in drug-resistant epilepsy. We aim to assess the localizing and lateralizing value as well as prognostic significance of fear aura in patients with focal epilepsy. METHODS: The occurrence of fear aura in relation to epileptogenic origin and its association with postoperative outcome were analyzed in 146 consecutive patients undergoing resective surgery for intractable epilepsy. RESULTS: Ninety-four (64.4%) patients reported auras, and 31 (21.2%) reported fear aura in their seizures. One hundred ten (75.3%) patients had an Engel class I outcome until last follow-up, of whom 24 experienced fear aura preoperatively. Fear aura appeared more frequently during temporal and frontal lobe seizures, but did not lateralize the seizure onset zone. There were no significant baseline differences between patients with and without fear aura. No correlation was found between postoperative outcome and the presence of auras. Occurrence of fear aura failed to show predictive value in surgical outcome whether in pooled or subgroup analysis. INTERPRETATION: This study advances our understanding of the origin of fear aura, and is helpful for presurgical evaluation and outcome prediction. Without lateralizing value, fear aura is more commonly seen with temporal or frontal origin. When taken as a whole, auras do not have a significant impact on seizure outcome in focal epilepsy. Patients with fear aura are no more likely to become seizure-free than those without fear aura.


Assuntos
Epilepsia Resistente a Medicamentos , Epilepsias Parciais , Epilepsia , Epilepsia Resistente a Medicamentos/cirurgia , Epilepsias Parciais/cirurgia , Medo , Humanos , Prognóstico , Convulsões , Lobo Temporal
12.
Spectrochim Acta A Mol Biomol Spectrosc ; 238: 118375, 2020 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-32438293

RESUMO

DNA methylation has been identified as one of the important causes of tumorigenesis, so it is important to develop some advanced methods for detecting and quantifying DNA methylation. In this study, a label-free and enzyme-free one-step rapid colorimetric detection of DNA methylation based on unmodified Au nanoparticles(Au NPs)has been proposed. This method can quickly, efficiently, economically and easily colorimetric detect methylated DNA only by the color change of unmodified Au NPs solution without the covalent modification of Au NPs in advance or complicated instruments for implementation with practical limitations or expensive biological enzymes or traditional organic dyes during the reaction. The strategy employed the difference in electrostatic attraction of single-stranded DNA and double-stranded DNA against salt-induced aggregation of Au NPs. The method has a DNA methylated detection limit of 8.47 nM and it is distinctly visible to detect methylated DNA with the naked eye as low as 20 nM. Furthermore, the strategy has an ability to detect methylated DNA in the presence of abundant unmethylated DNA with the detection limit of 0.13% and as low as 1% methylated DNA can be distinguished in heterogeneous samples with the naked eye. Also, the stratagem provides a convenient and rapid platform for methylated DNA detection of human serum samples in one step, which displays a huge potential for clinical diagnosis and treatment of oncological diseases.


Assuntos
Metilação de DNA , Ouro/química , Nanopartículas Metálicas/química , Colorimetria/economia , Colorimetria/métodos , DNA/sangue , DNA/química , Humanos , Fatores de Tempo
13.
Math Biosci Eng ; 17(3): 2496-2515, 2020 02 26.
Artigo em Inglês | MEDLINE | ID: mdl-32233551

RESUMO

Epilepsy is a brain syndrome caused by synchronous abnormal discharge of brain neurons. As an effective treatment for epilepsy, successful surgical resection requires accurate localization of epileptic foci to avoid iatrogenic disability. Previous studies have demonstrated the potential of restingstate functional magnetic resonance imaging (rs-fMRI) technique to localize epileptic foci though clinical applications of rs-fMRI are still at an early stage of development. fMRI data analysis approaches seek pre-defined regressors modeling contributions to the voxel time series, including the BOLD response following neuronal activation. In present study, localization strategies of epileptic foci in rs-fMRI technology were classified and summarized. To begin with, data-driven approaches attempting to determine the intrinsic structure of the data were discussed in detail. Then, as novel fMRI data analysis methods, deconvolution algorithms such as total activation (TA) and blind deconvolution were discussed, which were applied to explore the underlying activity-inducing signal of the BOLD signal. Lastly, effective connectivity approaches such as autocorrelation function method and Pearson correlation coefficient have also been proposed to identify the brain regions driving the generation of seizures within the epileptic network. In the future, fMRI technology can be used as a supplement of intraoperative subdural electrode method or combined with traditional epileptic focus localization technologies, which is one of the most attractive aspect in clinic. It may also play an important role in providing diagnostic information for epilepsy patients.


Assuntos
Mapeamento Encefálico , Epilepsia , Algoritmos , Encéfalo/diagnóstico por imagem , Eletroencefalografia , Epilepsia/diagnóstico por imagem , Humanos , Imageamento por Ressonância Magnética
14.
Curr Med Sci ; 39(6): 863-873, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31845216

RESUMO

Voltage-gated sodium (Nav) channels are critical players in the generation and propagation of action potentials by triggering membrane depolarization. Mutations in Nav channels are associated with a variety of channelopathies, which makes them relevant targets for pharmaceutical intervention. So far, the cryoelectron microscopic structure of the human Nav1.2, Nav1.4, and Nav1.7 has been reported, which sheds light on the molecular basis of functional mechanism of Nav channels and provides a path toward structure-based drug discovery. In this review, we focus on the recent advances in the structure, molecular mechanism and modulation of Nav channels, and state updated sodium channel blockers for the treatment of pathophysiology disorders and briefly discuss where the blockers may be developed in the future.


Assuntos
Canalopatias/genética , Mutação , Bloqueadores dos Canais de Sódio/química , Canais de Sódio Disparados por Voltagem/metabolismo , Potenciais de Ação , Canalopatias/tratamento farmacológico , Microscopia Crioeletrônica , Desenho de Fármacos , Humanos , Modelos Moleculares , Conformação Proteica , Bloqueadores dos Canais de Sódio/farmacologia , Bloqueadores dos Canais de Sódio/uso terapêutico , Relação Estrutura-Atividade , Canais de Sódio Disparados por Voltagem/química , Canais de Sódio Disparados por Voltagem/genética
15.
Chem Biol Interact ; 297: 119-129, 2019 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-30365938

RESUMO

Diabetes-associated sexual dysfunction and fertility impairments are major secondary complications in diabetic patients and animal models. Natural herbs are important sources of therapeutic agents for diabetic complications. This study investigated the effect of vitexin on male sexual dysfunction and fertility impairments in streptozotocin (STZ)-induced diabetic mice. Diabetes was induced by intraperitoneal injection of 45 mg/kg STZ for 5 consecutive days in mice. Vitexin (10, 20 or 40 mg/kg) and Sildenafil citrate (SC, 5 mg/kg) were administered daily for 62 days after the induction of diabetes. The parameters of sexual behavior and fertility were analyzed. The reproductive organ weight, sperm motility, and viability of the treated mice were examined. Testicular histopathological alterations were detected by hematoxylin and eosin (H&E) staining. Enzyme-linked immunosorbent assay (ELISA) was used to evaluate serum hormonal levels. Results showed that 40 mg/kg vitexin significantly improved the sexual behavior and fertility levels compared with the diabetic group. Moreover, vitexin (20 or 40 mg/kg) significantly increased reproductive organ weight and improved testicular pathological structure damage. Meanwhile, sperm analysis demonstrated that vitexin significantly restored sperm quality in a dose-dependent manner. Furthermore, ELISA data showed that vitexin significantly increased the serum testosterone (T), follicular-stimulating hormone (FSH), and luteinizing hormone (LH) levels but decreased the gonadotropin-releasing hormone (GnRH) level to different degrees. These findings suggest that vitexin ameliorates sexual dysfunction and fertility impairments in male diabetic mice possibly by modulating the hypothalamus-pituitary-gonadal axis.


Assuntos
Apigenina/farmacologia , Diabetes Mellitus Experimental/tratamento farmacológico , Fertilidade/efeitos dos fármacos , Gônadas/efeitos dos fármacos , Hipotálamo/efeitos dos fármacos , Hipófise/efeitos dos fármacos , Disfunções Sexuais Fisiológicas/tratamento farmacológico , Animais , Apigenina/administração & dosagem , Apigenina/química , Glicemia/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Diabetes Mellitus Experimental/induzido quimicamente , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Feminino , Sistema Hipotálamo-Hipofisário/efeitos dos fármacos , Injeções Intraperitoneais , Masculino , Camundongos , Estrutura Molecular , Disfunções Sexuais Fisiológicas/induzido quimicamente , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Estreptozocina/administração & dosagem , Relação Estrutura-Atividade
16.
Anal Chim Acta ; 1016: 12-18, 2018 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-29534800

RESUMO

In this work, a newly developed surface plasma resonance (SPR) system for the sensitive detection of M.SssI activity has been designed based on double signal amplification with DNA chain cyclic reactions and AuNPs. In the absence of M.SssI, hairpin DNA 1 (HP1) can be cleaved into s1 fragments catalyzed by HpaII. The s1 fragments can then trigger a recycling process of hairpin DNA 2 (HP2) hybridization and subsequently release massive s2 and s3 in the solution of Nt.AlwI and HPII. AuNPs-DNA can be captured on gold film by the released s2 and s3 to produce a strong SPR signal. Whereas in the presence of M.SssI, methylated HP1 cannot be cleaved by HpaII, thus produce a weak SPR signal. The SPR signals are dependent on the M.SssI concentration in the range from 0.5 to 50 U/mL. The successful detection of M.SssI activity in clinical serum samples and inhibition of M.SssI using 5-Aza and 5-Aza-dC indicate a great potential of this strategy for building new monitoring platform in bioanalysis and clinical biomedicine.


Assuntos
Metiltransferases/sangue , Ressonância de Plasmônio de Superfície , Técnicas Eletroquímicas , Ouro/química , Humanos , Metiltransferases/metabolismo , Propriedades de Superfície
17.
Anal Sci ; 34(8): 959-964, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30101892

RESUMO

DNA methyltransferase (MTase) is related to transcriptional repressor activity in biological functions. It is an essential for cancer diagnosis and therapeutics to detect DNA MTase activity sensitively. Here, a fluorescent system based on polymerase amplification has been developed to detect DNA adenine MTase (Dam) activity sensitively. The amplification is triggered by the probe DNA regions a, which are the primes of a polymerase-induced replicated reaction. They come from methylation and a digestion reaction of DNA S1-S1, including a 5'-GATC-3' sequence recognized by Dam MTase and methylation sensitive restriction endonuclease Dpn I. The intensities of fluorescence are dependent on the Dam MTase activity. The method shows fine sensitivity with a detection limit of 3.2 × 10-4 U mL-1 and specificity for Dam MTase. In human serum samples, the method has been successfully applied, and it has also been used to screen the inhibitors, which means that the developed method can be a powerful and potential tool for drug development and clinical diagnosis in the future.


Assuntos
Técnicas Biossensoriais , Ensaios Enzimáticos/métodos , Fluorescência , Técnicas de Amplificação de Ácido Nucleico/métodos , DNA Metiltransferases Sítio Específica (Adenina-Específica)/análise , DNA Metiltransferases Sítio Específica (Adenina-Específica)/metabolismo , Sondas de DNA/química , Sondas de DNA/metabolismo , Humanos , Espectrometria de Fluorescência
18.
Biosens Bioelectron ; 102: 211-216, 2018 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-29145074

RESUMO

We propose a ratiometric electrochemical assay for detecting microRNA (miRNA) on the basis of dual-amplification mechanism by using distinguishable electrochemical signals from thionine (Thi) and ferrocene (Fc). The thiol-modified and ferrocene-labeled hairpin capture probes (CP) are first immobilized on an Au electrode via Au-S reaction. The target miRNA hybridizes with CP and unfolding the hairpin structure of CP to form miRNA-DNA duplexes. Then, kamchatka crab duplex specific nuclease (DSN) specifically cleaves the DNA in miRNA-DNA duplexes, leading to the release of miRNA and another cleaves cycle, meanwhile, numerous Fc leaves away from the electrode surface and leads to the signal-off of Fc. The residual fragment on electrode surface acts as a HCR primer to form dsDNA polymers through in situ HCR with the presence of the primer and two probes (HDNA and HDNA'), resulting in the capture of numerous DNA/Au NPs/Thi and the signal-on of Thi. The dual-amplification mechanism significantly amplifies the decrease of Fc signal and the increase of Thi signal for ratiometric readout (IThi/IFc), thus providing a sensitive method for the selective detection of miR-141 with a detection limit down to 11aM. The dual-signal ratiometric outputs have an intrinsic self-calibration to the effects from system, which is promising to be applied in biosensing and clinical diagnosis.


Assuntos
Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Nanopartículas Metálicas/química , MicroRNAs/isolamento & purificação , Compostos Ferrosos/química , Ouro/química , Humanos , Limite de Detecção , Metalocenos/química , MicroRNAs/química , Hibridização de Ácido Nucleico , Fenotiazinas/química
19.
Biomed Pharmacother ; 90: 562-574, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28407577

RESUMO

Diabetes mellitus (DM) is a major health problem that affects patients' quality of life quality throughout the world due to its many complications. Reproductive dysfunction is one of the major secondary complications in both diabetic animals and human beings. Furthermore, DM has recently broken the age barrier and has been heavily diagnosed in children and young persons of reproductive age. In the past few years, many studies on DM in male reproductive functions in both diabetic men and experimental diabetic animals have been published. It is recognized that sustained hyperglycemia, which impairs reproductive function in diabetic men, is at risk of developing. DM harmfully affects male reproductive functions in multiple areas; these may include spermatogenesis, sperm maturation, fertility capability, penile erection, and ejaculation. Traditional medicine and folklore worldwide have used numerous medicinal plants to manage the diabetic reproductive dysfunction because bioactive phyto-constituents are affluent in many places. Unfortunately, the exact reasons for diabetic male reproductive dysfunction are not completely understood and currently there are no treatments in reproductive medicine specifically for such lesions. The aim of this review is to summarize current research findings of DM on reproductive functions, to elaborate the underlying mechanisms related to these diseases via in vivo and in vitro studies, and to describe the ameliorative effects of medicinal plants or their products. The review findings provide a systematic understanding of DM on the reproductive functions and lay the theoretical foundation for developing the direction of reproductive medicine.


Assuntos
Complicações do Diabetes/etiologia , Complicações do Diabetes/fisiopatologia , Diabetes Mellitus/fisiopatologia , Genitália Masculina/fisiopatologia , Animais , Humanos , Masculino
20.
Zhonghua Yi Xue Za Zhi ; 86(47): 3324-7, 2006 Dec 19.
Artigo em Zh | MEDLINE | ID: mdl-17313826

RESUMO

OBJECTIVE: To study the sub-classification of temporal lobe epilepsy (TLE) and clinical and etiological characteristics thereof. METHODS: The clinical data of 190 patients with TLE, aged 1.5 approximately 67, were analyzed. The sub-classification was based on clinical manifestations and electroencephalographic recording. The etiology of TLE was mainly concluded from both clinical history and neuroimaging. RESULTS: 172 patients were subclassified as with mesial temporal lobe epilepsy (MTLE), of which 62.2% had auras such as epigastric abnormal sensation and 67.4% had automatism, especially oroalimentary automatism. The electroencephalography recording of the MTLE patients showed interictal epileptic paroxysm discharges over the anterior or mid-anterior temporal areas. The main etiological factors related to MTLE were hippocampal sclerosis, intracranial infection, tumor, cerebrovascular disease, and trauma. Ten patients were subclassified as with lateral temporal lobe epilepsy (LTLE), all presenting auras such as acoust. The electroencephalography recording showed interictal epileptic paroxysm discharges over the posterior or mid-posterior temporal areas. The main etiological factors were trauma, tumor, and atrophy in the LTLE group. Eight patients were unable to be sub-classified. CONCLUSION: Electroencephalography is the main base for classification of TLE. Subclassification of TLE is important in the selection of treatment strategies.


Assuntos
Epilepsia do Lobo Temporal/classificação , Epilepsia do Lobo Temporal/fisiopatologia , Adolescente , Adulto , Distribuição por Idade , Idoso , Criança , Pré-Escolar , Eletroencefalografia , Epilepsia do Lobo Temporal/etiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
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