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1.
Cell Death Dis ; 11(4): 293, 2020 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-32341357

RESUMO

Non-traumatic osteonecrosis of the femoral head (ONFH) is clinically a devastating and progressive disease without an effective treatment. Mesenchymal stem cells (MSCs) transplantation has been used to treat ONFH in early stage, but the failure rate of this therapy is high due to the reduced osteogenic differentiation and migration of the transplanted MSCs related with pathological bone tissues. However, the mechanism responsible for this decrease is still unclear. Therefore, we assume that the implanted MSCs might be influenced by signals delivered from pathological bone tissue, where the exosomes might play a critical role in this delivery. This study showed that exosomes from ONFH bone tissues (ONFH-exos) were able to induce GC-induced ONFH-like damage, in vivo and impair osteogenic differentiation and migration of MSCs, in vitro. Then, we analyzed the differentially expressed proteins (DEPs) in ONFH-exos using proteomic technology and identified 842 differentially expressed proteins (DEPs). On the basis of gene ontology (GO) enrichment analysis of DEPs, fold-changes and previous report, cell adhesion-related CD41 (integrin α2b) was selected for further investigation. Our study showed that the CD41 (integrin α2b) was distinctly decreased in ONFH-exos, compared to NOR-exos, and downregulation of CD41 could impair osteogenic differentiation and migration of the MSCs, where CD41-integrin ß3-FAK-Akt-Runx2 pathway was involved. Finally, our study further suggested that CD41-affluent NOR-exos could restore the glucocorticoid-induced decline of osteogenic differentiation and migration in MSCs, and prevent GC-induced ONFH-like damage in rat models. Taken together, our study results revealed that in the progress of ONFH, exosomes from the pathological bone brought about the failure of MSCs repairing the necrotic bone for lack of some critical proteins, like integrin CD41, and prompted the progression of experimentally induced ONFH-like status in the rat. CD41 could be considered as the target of early diagnosis and therapy in ONFH.

2.
Parasitology ; : 1-9, 2020 Feb 12.
Artigo em Inglês | MEDLINE | ID: mdl-32046796

RESUMO

The rhoptry kinase 18 of Toxoplasma gondii (TgROP18) has been identified as a key virulence factor that allows the parasite to escape from host immune defences and promotes its proliferation in host cells. Although much research is focused on the interaction between host cells and TgROP18, the development of monoclonal antibodies (mAbs) against TgROP18 has not been reported till date. To produce mAbs targeting TgROP18, two hybridomas secreting mAbs against TgROP18, designated as A1 and T2, were generated using cell fusion technology. The subtypes of the A1 and T2 mAbs were identified as IgG3 λ and IgM κ, and peptide scanning revealed that the core sequences of the antigenic epitopes were 180LRAQRRRSELVFE192 and 351NYFLLMMRAEADM363, respectively. The T2 mAb specifically reacted with both T. gondii type I and Chinese I, but not with T. gondii type II, Plasmodium falciparum or Schistosoma japonicum. Finally, the sequences of heavy chain and light chain complementarity-determining regions of T2 were amplified, cloned and characterized, making the modification of the mAb feasible in the future. The development of mAbs against TgROP18 would aid the investigation of the molecular mechanisms underlying the modulation of host cellular functions by TgROP18, and in the development of strategies to diagnose and treat Toxoplasmosis.

3.
J Org Chem ; 85(4): 2532-2542, 2020 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-31910622

RESUMO

The copper-catalyzed [4 + 2] annulation of α,ß-unsaturated ketoxime acetates with 1,3-dicarbonyl compounds for the synthesis of three classes of structurally diverse pyridines has been developed. This method employs 1,3-dicarbonyl compounds as C2 synthons and enables the synthesis of multifunctionalized pyridines with diverse electron-withdrawing groups in moderate to good yields. The mechanistic investigation suggests that the reactions proceed through an ionic pathway.

4.
Biosens Bioelectron ; 148: 111826, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31707327

RESUMO

In this study, a multiple-legged and highly integrated DNA rolling walker based electrochemical biosensor was developed for ultrasensitive ctDNA analysis through rolling circle amplification (RCA) with doxorubicin@tetrahedron-Au (DOX@TDN-Au) as electrochemical indicator. Upon target-driven RCA, the multiple-legged walker could move along with the predesigned track by strand displacement reactions, resulting in numbers of legs binding irreversibly to iStep probes. The binding of massive legs to iStep probes could effectively impede DOX@TDN-Au tags binding on the surface of sensor and then reached a "signal off" state. Benefiting from the highly amplified efficiency of rolling walker machine and DOX@TDN-Au tags, the established biosensor performed high sensitivity for target detection with a low limit of detection down to 0.29 fM. Moreover, the target ctDNA could hybridize with the ring and capture probe simultaneously, greatly enhancing the specificity of the developed biosensing method. Thus, this biosensing method is a promising tool for detection of ctDNA in the field of clinical diagnostic and tumor progression assessment.

5.
Cell Metab ; 30(1): 157-173.e7, 2019 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-31155494

RESUMO

We report that circACC1, a circular RNA derived from human ACC1, plays a critical role in cellular responses to metabolic stress. CircACC1 is preferentially produced over ACC1 in response to serum deprivation by the transcription factor c-Jun. It functions to stabilize and promote the enzymatic activity of the AMPK holoenzyme by forming a ternary complex with the regulatory ß and γ subunits. The cellular levels of circACC1 modulate both fatty acid ß-oxidation and glycolysis, resulting in profound changes in cellular lipid storage. In a tumor xenograft model, silencing or enforced expression of circACC1 resulted in growth inhibition and enhancement, respectively. Moreover, increased AMPK activation in colorectal cancer tissues was frequently associated with elevated circACC1 expression. We conclude that circACC1 serves as an economic means to elicit AMPK activation and moreover propose that cancer cells exploit circACC1 during metabolic reprogramming.

6.
Oncol Rep ; 41(6): 3189-3200, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31002363

RESUMO

Actin-related protein 2/3 complex (ARPC2) is an actin­binding component involved in the regulation of actin polymerization. It mediates the formation of branched actin networks and contacts the mother actin filament. Migration and invasion are key processes which enable tumor cells to infiltrate blood vessels or lymphatic vessels, and the actin pathway plays a very important role. Given that ARPC2 is critical to this progression, the present study focused on ARPC2 activity in breast cancer (BrCa) cell invasion and migration. Limited data are available on the expression and role of ARPC2 proteins in breast carcinomas. We screened the Oncomine database for messenger RNAs (mRNAs) that are upregulated in BrCa and found that ARPC2 was one of the most consistently involved mRNAs in BrCa. The analysis of immunohistochemical data revealed that ARPC2 expression was higher in breast cancerous tissues than in adjacent non­cancerous tissues. In addition, ARPC2 was highly associated with the tumor stage, nodal metastasis, and overall survival of patients with BrCa. We performed siRNA­ARPC2 transfection to investigate the effect of ARPC2 on the proliferation, migration, invasion and arrest of BrCa cells. It was revealed that ectopic ARPC2 expression significantly upregulated N­cadherin, vimentin, ZEB1, MMP­9 and MMP­3 expression and also activated the TGF­ß pathway to contribute to epithelial­mesenchymal transition (EMT). These results collectively indicated that ARPC2 promoted the tumorigenesis of breast carcinoma and the initiation of EMT. Therefore, ARPC2 was revealed to be a potential therapeutic target in patients with BrCa.


Assuntos
Complexo 2-3 de Proteínas Relacionadas à Actina/genética , Neoplasias da Mama/genética , Carcinogênese/genética , Proliferação de Células/genética , Adulto , Neoplasias da Mama/patologia , Neoplasias da Mama/terapia , Linhagem Celular Tumoral , Movimento Celular/genética , Transição Epitelial-Mesenquimal/genética , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Pessoa de Meia-Idade , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , Metástase Neoplásica , RNA Mensageiro/genética , RNA Interferente Pequeno/genética , Transfecção
7.
Onco Targets Ther ; 11: 6911-6924, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30410349

RESUMO

Background: Colorectal cancer stem cells (CRC-SCs) contribute to the initiation and progression of colorectal cancer (CRC). However, the underlying mechanisms for the propagation of CRC-SCs have remained elusive. Purpose: The objective of this study was to study the role of NFATC2 in maintenance of the stemness in CRC-SCs. Method: The expression levels of mRNA and protein were determined by qRT-PCR and western-blot, respectively. CRC-SCs were isolated by spheroid formation assay and flowcytometry. The sphere-forming and self-renewal abilities of CRC-SCs were determined by spheroid formation assay. The tumorigenicity of CRC-SCs was determined by cell-derived xenograft model. Gene manipulation was performed by lentivirus-mediated delivery system. Results: We first found that NFATC2 is upregulated in primary CRC-SCs. Overexpression of NFATC2 promotes self-renewal and the expression of stem cell markers of CRC-SCs. Conversely, knockdown of NFATC2 attenuates stem cell-like properties of CRC-SCs. Mechanistic analysis indicated that NFATC2 upregulates the expression of AJUBA, downregulates the phosphorylation level of YAP, and therefore activates the transcriptional activities of YAP and promotes the stemness of CRC-SCs. Conclusion: Our findings demonstrate NFATC2 as an oncogene that can promote the stemness of CRC-SCs. This work suggests a novel therapeutic strategy against CRC caused by aberrant expression of NFATC2.

8.
Artigo em Inglês | MEDLINE | ID: mdl-30333962

RESUMO

Biofilm formation is critical for blocking flea foregut and hence for transmission of Y. pestis by flea biting. In this study, we identified the regulatory role of the AraC-family transcriptional regulator BfvR (YPO1737 in strain CO92) in biofilm formation and virulence of Yersinia pestis biovar Microtus. Crystal violet staining, Caenorhabditis elegans biofilm assay, colony morphology assay, intracellular c-di-GMP concentration determination, and BALB/c mice challenge were employed to reveal that BfvR enhanced Y. pestis biofilm formation while repressed its virulence in mice. Further molecular biological assays demonstrated that BfvR directly stimulated the expression of hmsHFRS, waaAE-coaD, and hmsCDE, which, in turn, affected the production of exopolysaccharide, LPS, and c-di-GMP, respectively. In addition, BfvR directly and indirectly repressed psaABC and psaEF transcription, respectively. We concluded that the modulation of biofilm- and virulence-related genes by BfvR led to increased biofilm formation and reduced virulence of Y. pestis biovar Microtus.


Assuntos
Antígenos de Bactérias/metabolismo , Biofilmes/crescimento & desenvolvimento , Regulação Bacteriana da Expressão Gênica , Fatores de Transcrição/metabolismo , Yersinia pestis/crescimento & desenvolvimento , Yersinia pestis/metabolismo , Animais , Caenorhabditis elegans/microbiologia , GMP Cíclico/análogos & derivados , GMP Cíclico/análise , Modelos Animais de Doenças , Redes Reguladoras de Genes , Genes Reguladores , Camundongos Endogâmicos BALB C , Peste/microbiologia , Peste/patologia , Polissacarídeos Bacterianos/metabolismo , Análise de Sobrevida , Virulência , Yersinia pestis/genética
9.
Analyst ; 142(24): 4834-4842, 2017 Dec 04.
Artigo em Inglês | MEDLINE | ID: mdl-29160870

RESUMO

Recently, hairpin stacking circuits (HSC) based on toehold-mediated strand displacement have been engineered to detect nucleic acids and proteins. However, the three metastable hairpins in a HSC system can potentially react non-specifically in the absence of a catalyst, limiting its practical application. Here, we developed a unique hairpin design guideline to eliminate circuit leakage of HSC, and the high-performance HSC was successfully implemented on logic gate building and biosensing. We began by analyzing the sources of circuit leakage and optimizing the toehold lengths of hairpins in the HSC system based on the surface plasmon resonance (SPR) technique. Next, a novel strategy of substituting two nucleotides in a specific domain, termed 'loop-domain substitution', was introduced to eliminate leakages. We also systematically altered the positions and numbers of the introduced substitutions to probe their potential contribution to circuit leakage suppression. Through these efforts, the circuit leakage of HSC was significantly reduced. Finally, by designing different DNA input strands, the logic gates could be activated to achieve the output signal. Using miRNA as a model analyte, this strategy could detect miRNA down to pM levels with minimized circuit leakage. We believe these work indicate significant progress in the DNA circuitry.


Assuntos
Técnicas Biossensoriais , MicroRNAs/análise , Ressonância de Plasmônio de Superfície , DNA , Lógica
10.
Sci Rep ; 7(1): 14037, 2017 10 25.
Artigo em Inglês | MEDLINE | ID: mdl-29070911

RESUMO

In this work, a simple and enzyme-free surface plasmon resonance (SPR) biosensing strategy has been developed for highly sensitive detection of two major PML/RARα (promyelocytic leukemia, retinoic acid receptor alpha) subtypes based on the heterogeneous fusion gene-triggered nonlinear hybridization chain reaction (HCR). On the gold chip surface, the cascade self-assembly process is triggered after the introduction of PML/RARα. The different fragments of PML/RARα can specifically hybridize with capture probes (Cp) immobilized on the chip and the hybridization DNA1 (H1). Then, the nonlinear HCR is initiated by the complex of Cp-PML/RARα-H1 with the introduction of two hybridization DNA chains (H2 and H3). As a result, a dendritic nanostructure is achieved on the surface of chip, leading to a significant SPR amplification signal owing to its high molecular weight. The developed method shows good specificity and high sensitivity with detection limit of 0.72 pM for "L" subtype and 0.65 pM for "S" subtype. Moreover, this method has been successfully applied for efficient identification of clinical positive and negative PCR samples of the PML/RARα subtype. Thus, this developed biosensing strategy presents a potential platform for analysis of fusion gene and early diagnosis of clinical disease.

11.
Biosens Bioelectron ; 87: 345-351, 2017 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-27587359

RESUMO

A label-free and enzyme-free surface plasmon resonance (SPR) biosensing strategy has been developed for highly sensitive and specific detection of target DNA by employing the nonlinear hybridization chain reaction (HCR) amplification. Nonlinear HCR is a hairpin-free system in which double-stranded DNA monomers could dendritically assemble into highly branched nanostructure upon introducing a trigger sequence. The target DNA partly hybridizes with capture probe on the gold sensing chip and the unpaired fragment of target DNA works as a trigger to initiate the nonlinear HCR, forming a chain-branching growth of DNA dendrimer by self-assembly. Real-time amplified SPR response is observed upon the introduction of nonlinear HCR system. The method is capable of detecting target DNA at the concentration down to 0.85 pM in 60min with a dynamic range from 1 pM to 1000 pM, and could discriminate target DNA from mismatched sequences. This biosensing strategy exhibits good reproducibility and precision, and has been successfully applied for detection of target DNA in complex sample matrices. In addition, the nonlinear HCR based SPR biosensing methodology is extended to the detection of adenosine triphosphate (ATP) by aptamer recognition. Thus, the versatile method might become a potential alternative tool for biomolecule detection in medical research and early clinical diagnosis.


Assuntos
Trifosfato de Adenosina/análise , DNA/análise , Hibridização de Ácido Nucleico/métodos , Ressonância de Plasmônio de Superfície/métodos , Aptâmeros de Nucleotídeos/química , Sondas de DNA/química , Ouro/química , Nanoestruturas/química , Reprodutibilidade dos Testes
12.
Biochem Biophys Res Commun ; 473(1): 187-193, 2016 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-27012204

RESUMO

Toxoplasma gondii is a major cause of congenital brain disease. T. gondii infection in the developing fetus frequently results in major neural developmental damage; however, the effects of the parasite infection on the neural stem cells, the key players in fetal brain development, still remain elusive. This study is aiming to explore the role of T. gondii infection on differentiation of neural stem cells (NSCs) and elucidate the underlying molecular mechanisms that regulate the inhibited differentiation of NSCs induced by the infection. Using a differentiation medium, i.e. , DMEM: F12 (1:1 mixture) supplemented with 2% N2, C17.2 neural stem cells (NSCs) were able to differentiate to neurons and astrocytes, respectively evidenced by immunofluorescence staining of differentiation markers including ßIII-tubulin and glial fibrillary acidic protein (GFAP). After 5-day culture in the differentiation medium, the excreted-secreted antigens of T. gondii (Tg-ESAs) significantly down-regulated the protein levels of ßIII-tubulin and GFAP in C17.2 NSCs in a dose-dependent manner. The protein level of ß-catenin in the nucleus of C17.2 cells treated with both wnt3a (a key activator for Wnt/ß-catenin signaling pathway) and Tg-ESAs was significantly lower than that in the cells treated with only wnt3a, but significantly higher than that in the cells treated with only Tg-ESAs. In conclusion, the ESAs of T. gondii RH blocked the differentiation of C17.2 NCSs and downregulated the expression of ß-catenin, an essential component of Wnt/ß-catenin signaling pathway. The findings suggest a new mechanism underlying the neuropathogenesis induced by T. gondii infection, i.e. inhibition of the differentiation of NSCs via blockade of Wnt/ß-catenin signaling pathway, such as downregulation of ß-catenin expression by the parasite ESAs.


Assuntos
Células-Tronco Neurais/citologia , Células-Tronco Neurais/parasitologia , Toxoplasma , Proteínas Wnt/metabolismo , Via de Sinalização Wnt , beta Catenina/metabolismo , Animais , Antígenos/química , Diferenciação Celular , Núcleo Celular/metabolismo , Proliferação de Células/efeitos dos fármacos , Regulação para Baixo , Proteína Glial Fibrilar Ácida/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Camundongos , Microscopia de Fluorescência , Neurônios/metabolismo
13.
Biosens Bioelectron ; 80: 98-104, 2016 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-26807523

RESUMO

MicroRNAs (miRNAs) are potentially useful biomarkers for early diagnosis of human diseases. Here, a simple surface plasmon resonance (SPR) biosensor has been developed for highly sensitive detection of miRNA by designing a new enzyme-free and isothermal amplification strategy, named multi component nucleic acid enzyme-mediated mismatched catalyzed hairpin assembly (MNAzyme-CHA). The partial MNAzymes co-recognized the target to form a stable active MNAzyme, which continued to digest multiple hairpin H0 substrates, concomitantly generating a lot of fragments. The H0 fragments could initiate the mismatched CHA cycles, resulting in the generation of massive hairpin H1-H2 complexes. As a result, the H1-H2 complexes and streptavidin were attached to the sensor surface, leading to a significantly amplified SPR signal readout. The established biosensor showed high sensitivity and selectivity with a wide dynamic range from 1 pM to 100 nM. It was also successfully applied to the determination of target miRNA spiked into human total RNA samples. Thus, this developed biosensing strategy presents a simple and stable platform toward sensitive and convenient miRNA detection, and has great potential in assays of many other nucleic acids analytes for biomedical research and early clinical diagnosis.


Assuntos
Técnicas Biossensoriais/métodos , MicroRNAs/isolamento & purificação , Conformação de Ácido Nucleico , Ácidos Nucleicos/química , Catálise , Humanos , Limite de Detecção , MicroRNAs/química , Estreptavidina/química , Ressonância de Plasmônio de Superfície
14.
Anal Chim Acta ; 874: 59-65, 2015 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-25910447

RESUMO

MicroRNAs (miRNAs) play an important regulatory role in cells and dysregulation of miRNA has been associated with a variety of diseases, making them a promising biomarker. In this work, a novel biosensing strategy has been developed for label-free detection of miRNA using surface plasmon resonance (SPR) coupled with DNA super-sandwich assemblies and biotin-strepavidin based amplification. The target miRNA is selectively captured by surface-bound DNA probes. After hybridization, streptavidin is employed for signal amplification via binding with biotin on the long DNA super-sandwich assemblies, resulting in a large increase of the SPR signal. The method shows very high sensitivity, capable of detecting miRNA at the concentration down to 9 pM with a wide dynamic range of 6 orders of magnitude (from 1 × 10(-11) M to 1 × 10(-6) M) in 30 min, and excellent specificity with discriminating a single base mismatched miRNA sequence. This biosensor exhibits good reproducibility and precision, and has been successfully applied to the detection of miRNA in total RNA samples extracted from human breast adenocarcinoma MCF-7 cells. It, therefore, offers a highly effective alternative approach for miRNA detection in biomedical research and clinical diagnosis.


Assuntos
Ácidos Nucleicos Imobilizados/química , MicroRNAs/análise , Estreptavidina/química , Pareamento Incorreto de Bases , Sondas de DNA/química , Humanos , Limite de Detecção , Células MCF-7 , MicroRNAs/genética , Hibridização de Ácido Nucleico , Ressonância de Plasmônio de Superfície/métodos
15.
Parasit Vectors ; 8: 73, 2015 Feb 04.
Artigo em Inglês | MEDLINE | ID: mdl-25649541

RESUMO

BACKGROUND: Toxoplasma gondii, an obligate intracellular pathogen, has a strong affinity for the nervous system. TgCtwh3, a representative Chinese 1 Toxoplasma strain prevalent in China, has the polymorphic features of the effectors ROP16I/III with type I and GRA15II with type II Toxoplasma strains. The interaction of this atypical strain with host cells remains extremely elusive. METHODS: Using a transwell system, neural stem cells C17.2 were co-cultured with the tachyzoites of TgCtwh3 or standard type I RH strain. The apoptosis levels of C17.2 cells and the expression levels of related proteins in the endoplasmic reticulum stress (ERS)-mediated pathway were detected by flow cytometry and Western blotting. RESULTS: The apoptosis level of C17.2 cells co-cultured with TgCtwh3 had a significant increase compared to the negative control group; however, the apoptosis level in the TgCtwh3 group was significantly lower than that in the RH co-culture group. Western blotting analyses reveal that, after the C17.2 cells were co-cultured with TgCtwh3 and RH tachyzoites, the expression levels of caspase-12, CHOP and p-JNK in the cells increased significantly when compared to the control groups. After the pretreatment of Z-ATAD-FMK, an inhibitor of caspase-12, the apoptosis level of the C17.2 cells co-cultured with TgCtwh3 or RH tachyzoites had an apparent decline, and correspondingly, the expression levels of those related proteins were notably decreased. CONCLUSIONS: Our findings suggest that TgCtwh3 may induce the apoptosis of the C17.2 cells by up-regulation of caspase-12, CHOP, and p-JNK, which are associated with ERS signaling pathways. This work contributes to better understanding the possible mechanism of brain pathology induced by T. gondii Chinese 1 isolates prevalent in China, and also reveals the potential value of ERS inhibitors to treat such related diseases in the future.


Assuntos
Apoptose , Estresse do Retículo Endoplasmático , Células-Tronco Neurais/citologia , Toxoplasma/fisiologia , Toxoplasmose/metabolismo , Animais , Caspase 12/genética , Caspase 12/metabolismo , China , Humanos , Camundongos , Células-Tronco Neurais/metabolismo , Células-Tronco Neurais/parasitologia , Transdução de Sinais , Toxoplasmose/parasitologia , Toxoplasmose/fisiopatologia , Fator de Transcrição CHOP/genética , Fator de Transcrição CHOP/metabolismo
16.
Biosens Bioelectron ; 68: 343-349, 2015 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-25603399

RESUMO

MicroRNAs (miRNAs) play vital regulatory roles in cancer development and a variety of diseases, which make them become promising biomarkers. Here, a simple electrochemical biosensor was developed for highly sensitive and specific detection of target miRNA using mismatched catalytic hairpin assembly (CHA). The target miRNA triggered the toehold strand displacement assembly of two hairpin substrates, which led to the cyclic reuse of the target miRNA and the CHA products. Compared with the traditional CHA, mismatched CHA could decrease the nonspecific CHA products, which reduced the background signal significantly. Under the optimal experimental conditions and using differential pulse voltammetry, the established biosensor could detect target miRNA down to 0.6 pM (S/N=3) with a linear range from 1 pM to 25 nM, and discriminate target miRNA from mismatched miRNA with a high selectivity. It was also applied to the determination of miRNA spiked into human total RNA samples. Thus, this biosensing strategy might become a potential alternative tool for detection of miRNA in biomedical research and early clinical diagnosis.


Assuntos
Técnicas Eletroquímicas/instrumentação , MicroRNAs/análise , Pareamento Incorreto de Bases , Técnicas Biossensoriais/instrumentação , Humanos , Limite de Detecção , MicroRNAs/genética
17.
Zhongguo Zhen Jiu ; 34(4): 347-9, 2014 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-24946633

RESUMO

OBJECTIVE: To explore the better therapy in the treatment of ganglion. METHODS: Ninety cases of ganglion were randomized into a two-way quintuple puncture group, a common quintuple puncture group and a fire needling group, 30 cases in each one. In the two-way quintuple puncture group, the "9-in-1" multiple penetrating needling technique was used. In the common quintuple puncture group, the traditional "5-in-1" multiple penetrating needling technique was applied. In the fire needling group, the traditional multiple fire needling technique was adopted. The treatment was given once a day, 3 treatments made one session and the efficacy was analyzed statistically after 1 session treatment in the three groups. RESULTS: All of the three therapeutic methods achieved the efficacy on ganglion. The curative rate was 96. 7% (29/30) in the two-way quintuple puncture group, which was better obviously than 66.7% (20/30) in the common quintuple puncture group and 60. 0% (18/30) in the fire needling group (both P<0. 01). CONCLUSION: The two-way quintuple puncture technique achieves the remarkably superior efficacy on ganglion as compared with the common quintuple puncture technique and fire needling technique.


Assuntos
Terapia por Acupuntura , Cistos Glanglionares/terapia , Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Resultado do Tratamento , Adulto Jovem
18.
Parasitology ; 141(7): 988-95, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24612639

RESUMO

Toxoplasma gondii is a major cause of congenital brain disease; however, the underlying mechanism of neuropathogenesis in brain toxoplasmosis remains elusive. To explore the role of T. gondii in the development of neural stem cells (NSCs), NSCs were isolated from GD14 embryos of ICR mice and were co-cultured with tachyzoites of T. gondii RH strain. We found that apoptosis levels of the NSCs co-cultured with 1×106 RH tachyzoites for 24 and 48 h significantly increased in a dose-dependent manner, as compared with the control. Western blotting analysis displayed that the protein level of C/EBP homologous protein (CHOP) was up-regulated, and caspase-12 and c-Jun N-terminal kinase (JNK) were activated in the NSCs co-cultured with the parasites. Pretreatment with endoplasmic reticulum stress (ERS) inhibitor (TUDCA) and caspase-12 inhibitor (Z-ATAD-FMK) inhibited the expression or activation of the key molecules involved in the ERS-mediated apoptotic pathway, and subsequently decreased the apoptosis levels of the NSCs induced by the T. gondii. The findings here highlight that T. gondii induced apoptosis of the NSCs through the ERS signal pathway via activation of CHOP, caspase-12 and JNK, which may constitute a potential molecular mechanism responsible for the cognitive disturbance in neurological disorders of T. gondii.


Assuntos
Proteínas Reguladoras de Apoptose/fisiologia , Retículo Endoplasmático/fisiologia , Proteínas Mitocondriais/fisiologia , Células-Tronco Neurais/fisiologia , Toxoplasma/fisiologia , Animais , Técnicas de Cocultura , Embrião de Mamíferos , Feminino , Masculino , Camundongos , Camundongos Endogâmicos ICR , Estresse Fisiológico
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