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1.
Medicine (Baltimore) ; 100(27): e26417, 2021 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-34232173

RESUMO

BACKGROUND: Currently, there are a number of sodium glucose co-transport-2 (SGLT2) inhibitors that are under development or in clinical trials. Prior meta-analyses had established the safety and efficacy of SGLT2 inhibitors in type 1 diabetes mellitus (T1DM), but with low level of evidences and inconsistent conclusions. However, recently many new randomized clinical trials (RCTs) have been published, we hence try to design a study protocol to assess the effect of SGLT2 inhibitors on cardiovascular events via a comprehensive meta-analysis of data from much more RCTs, including sensitivity and subgroup analyses. METHODS: We will follow the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) reporting guidelines to conduct this meta-analysis. Two investigators will perform a systematic search of scientific literature in the databases (from conception through June 12, 2021), including PubMed, Embase, and Cochrane Central Register of Controlled Trials. This meta-analysis will be conducted using RevMan statistical software. The risk of bias for each included study will be assessed using the Cochrane Risk of Bias Assessment Tool. RESULTS: Our protocol is conceived to test the hypothesis that SGLT2 inhibitors could lead to better outcomes in patients presenting with T1DM. REGISTRATION NUMBER: 10.17605/OSF.IO/ZD8WX.


Assuntos
Diabetes Mellitus Tipo 1/tratamento farmacológico , Hipoglicemiantes/uso terapêutico , Inibidores do Transportador 2 de Sódio-Glicose/uso terapêutico , Diabetes Mellitus Tipo 1/sangue , Controle Glicêmico , Humanos
2.
eNeuro ; 8(2)2021.
Artigo em Inglês | MEDLINE | ID: mdl-33785521

RESUMO

Circuit compensation is often observed in patients with acute ischemic stroke, suggesting the importance of the interaction between brain regions. Also, contextual fear memory is an association between multisensory contexts and fearful stimuli, for which the interaction between the hippocampus and the amygdala is believed to be critical. To understand how focal ischemia in one region could influence the other region, we used a modified photo-thrombosis to induce focal ischemia in the hippocampus or the amygdala or both in freely-moving rats. We found that the learning curve and short-term memory (STM) were not affected in the rats although focal ischemia was induced 5 h before learning in either the hippocampus or the amygdala; these were impaired by the induction of ischemia in both the regions. Furthermore, the learning curve and STM were impaired when ischemia was induced 24 h before learning in either the hippocampus or the amygdala when the synaptic transmission was altered in one region because of ischemia in the other region. These results suggest that the circuit compensation between the hippocampus and the amygdala is critical for fear memory acquisition.


Assuntos
Isquemia Encefálica , Acidente Vascular Cerebral , Tonsila do Cerebelo , Animais , Medo , Hipocampo , Humanos , Isquemia , Ratos
3.
Neurotherapeutics ; 18(2): 1064-1080, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33786807

RESUMO

Brain capillaries are crucial for cognitive functions by supplying oxygen and other nutrients to and removing metabolic wastes from the brain. Recent studies have demonstrated that constriction of brain capillaries is triggered by beta-amyloid (Aß) oligomers via endothelin-1 (ET1)-mediated action on the ET1 receptor A (ETRA), potentially exacerbating Aß plaque deposition, the primary pathophysiology of Alzheimer's disease (AD). However, direct evidence is still lacking whether changes in brain capillaries are causally involved in the pathophysiology of AD. Using APP/PS1 mouse model of AD (AD mice) relative to age-matched negative littermates, we identified that reductions of density and diameter of hippocampal capillaries occurred from 4 to 7 months old while Aß plaque deposition and spatial memory deficit developed at 7 months old. Notably, the injection of ET1 into the hippocampus induced early Aß plaque deposition at 5 months old in AD mice. Conversely, treatment of ferulic acid against the ETRA to counteract the ET1-mediated vasoconstriction for 30 days prevented reductions of density and diameter of hippocampal capillaries as well as ameliorated Aß plaque deposition and spatial memory deficit at 7 months old in AD mice. Thus, these data suggest that reductions of density and diameter of hippocampal capillaries are crucial for initiating Aß plaque deposition and spatial memory deficit at the early stages, implicating the development of new therapies for halting or curing memory decline in AD.

4.
Biochem Biophys Res Commun ; 533(4): 1309-1314, 2020 12 17.
Artigo em Inglês | MEDLINE | ID: mdl-33051059

RESUMO

Spatial learning and memory are typically assessed to evaluate hippocampus-dependent cognitive and memory functions in vivo. Protein phosphorylation and dephosphorylation by kinases and phosphatases play critical roles in spatial learning and memory. Here we report that the Wip1 phosphatase is essential for spatial learning, with knockout mice lacking Wip1 phosphatase exhibiting dysfunctional spatial cognition. Aberrant phosphorylation of the Wip1 substrates p38, ATM, and p53 were observed in the hippocampi of Wip1-/- mice, but only p38 inhibition reversed impairments in long-term potentiation in Wip1-knockout mice. p38 inhibition consistently ameliorated the spatial learning dysfunction caused by Wip1 deficiency. Our results demonstrate that deletion of Wip1 phosphatase impairs hippocampus-dependent spatial learning and memory, with aberrant downstream p38 phosphorylation involved in this process and providing a potential therapeutic target.


Assuntos
Memória , Proteína Fosfatase 2C/fisiologia , Aprendizagem Espacial , Animais , Hipocampo/enzimologia , Hipocampo/fisiologia , Potenciação de Longa Duração , Masculino , Camundongos Knockout , Teste do Labirinto Aquático de Morris , Fosforilação , Proteína Fosfatase 2C/genética , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
5.
Front Pharmacol ; 11: 776, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32528295

RESUMO

Oxymatrine (OMT), a natural quinoxaline alkaloid extracted from the root of Sophora flavescens, presents amounts of pharmacological properties including immunomodulation, anti-inflammation, anti-oxidation, and anti-virus. Recent studies tend to focus on its effects on neuroinflammation and neuroprotection in Parkinson's disease (PD) due to its profound anti-inflammatory effect. In this study, the neuroprotective and anti-neuroinflammatory effects of OMT were investigated in 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP)-stimulated mice and 1-methyl-4-phenylpyridinium (MPP+)-induced mice primary microglia. Additionally, mice primary neuron-microglia co-cultures and primary microglia infected with Cathepsin D (CathD)-overexpressed lentivirus were used to clarify whether the neuroprotective effect of OMT was through a CathD-dependent pathway. Results showed that OMT dose-dependently alleviated MPTP-induced motor deficits and conferred significant dopamine (DA) neuroprotection against MPTP/MPP+-induced neurotoxicity. In addition, OMT inhibited MPTP/MPP+-induced microglia activation and the pro-inflammatory cytokines release. Further, OMT down-regulated the expression of CathD, and inhibited the activation of the HMGB1/TLR4 signaling pathway as well as the nuclear translocation of NF-κB both in vivo and in vitro. It is worth noting that overexpression of CathD reversed OMT-targeted inhibition of HMGB1/TLR4/NF-κB signaling and OMT-produced neuroprotection in reconstituted neuron-microglia co-cultures. Our findings indicated that OMT conferred DA neuroprotection and attenuated microglial-mediated neuroinflammation through CathD-dependent inhibition of HMGB1/TLR4/NF-κB signaling pathway. Our study supports a potential role for OMT in ameliorating PD, and proposes that OMT may be useful in the treatment of PD.

6.
Neurochem Res ; 45(5): 1107-1119, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32067150

RESUMO

miR-126 which is considered one of the most important miRNAs for maintaining vascular integrity, plays an important role in neuroprotection after cerebral ischemia-reperfusion (I-R). Moreover, vascular endothelial growth factor A (VEGFA), sprouty-related EVH1 domain-containing protein 1 (SPRED1), and Raf-1 are also involved in physiological processes of vascular endothelial cells (ECs). This study investigated how miR-126 changes with reperfusion time in different brain tissues after global cerebral ischemia and focal cerebral ischemia and examined the underlying mechanism miR-126 involving VEGFA, SPRED1, and Raf-1 after I-R. The results indicated decreases in the levels of miR-126-3p and miR-126-5p expression in mice and gerbils after I-R, consistent with the results after oxygen and glucose deprivation and reperfusion (OGD/R) in PC12 cells. Glial cells were activated as neuronal damage gradually increased after I-R. Inhibition of miR-126-3p exacerbated the OGD/R-induced cell death and reduced cell viability. After miR-126-3p inhibition, the levels of SPRED1 and VEGFA expression were increased, and p-Raf-1 expression was decreased after OGD/R. Moreover, based on the intervention of miR-126-3p inhibition, we found that the expression of p-Raf-1 was significantly increased after the intervention of siSPRED1, while it was not statistically significant after intervention of siVEGFA. The reduction of miR-126 expression after global and focal cerebral ischemia exacerbated neuronal death, which was closely related to increasing the SPRED1 activation and inhibiting the Raf-1 expression.


Assuntos
Isquemia Encefálica/metabolismo , Hipocampo/metabolismo , MicroRNAs/biossíntese , Traumatismo por Reperfusão/metabolismo , Animais , Isquemia Encefálica/genética , Isquemia Encefálica/patologia , Expressão Gênica , Gerbillinae , Hipocampo/patologia , Ataque Isquêmico Transitório/genética , Ataque Isquêmico Transitório/metabolismo , Ataque Isquêmico Transitório/patologia , Masculino , Camundongos , Camundongos Endogâmicos ICR , MicroRNAs/genética , Células PC12 , Ratos , Traumatismo por Reperfusão/genética , Traumatismo por Reperfusão/patologia
7.
Int J Mol Sci ; 20(20)2019 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-31614592

RESUMO

As organelles for photosynthesis in green plants, chloroplasts play a vital role in solar energy capture and carbon fixation. The maintenance of normal chloroplast physiological functions is essential for plant growth and development. Low temperature is an adverse environmental stress that affects crop productivity. Low temperature severely affects the growth and development of plants, especially photosynthesis. To date, many studies have reported that chloroplasts are not only just organelles of photosynthesis. Chloroplasts can also perceive chilling stress signals via membranes and photoreceptors, and they maintain their homeostasis and promote photosynthesis by regulating the state of lipid membranes, the abundance of photosynthesis-related proteins, the activity of enzymes, the redox state, and the balance of hormones and by releasing retrograde signals, thus improving plant resistance to low temperatures. This review focused on the potential functions of chloroplasts in fine tuning photosynthesis processes under low-temperature stress by perceiving stress signals, modulating the expression of photosynthesis-related genes, and scavenging excess reactive oxygen species (ROS) in chloroplasts to survive the adverse environment.


Assuntos
Cloroplastos/metabolismo , Estresse Fisiológico , Viridiplantae/crescimento & desenvolvimento , Ciclo do Carbono , Temperatura Baixa , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Fotossíntese , Proteínas de Plantas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Viridiplantae/metabolismo
8.
PLoS One ; 14(9): e0223228, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31557269

RESUMO

Chloroplast plays an important role in the plant life cycle. However, the details of its development remain elusive in rice. In this study, we report the fine-mapping of a novel rice gene wpb1 (white panicle branch 1), which affects chloroplast biogenesis, from a tropical japonica variety that results in an albino panicle branches at and after the heading stage. The wpb1 variety was crossed with Nipponbare to generate the F2 and BC1F2 populations. Green and white panicle branch phenotypes with a 3:1 segregation ratio was observed in the F2 population. Bulked segregant analysis (BSA) based on whole genome resequencing was conducted to determine the wpb1 locus. A candidate interval spanning from 11.35 to 23.79M (physical position) on chromosome 1 was identified. The results of BSA analysis were verified by a 40K rice SNP-array using the BC1F2 population. A large-scale F2 population was used to pinpoint wpb1, and the locus was further narrowed down to a 95-kb interval. Furthermore, our results showed that the expression levels of the majority of the genes involved in Chl biosynthesis, photosynthesis and chloroplast development were remarkably affected in wpb1 variety and in F2 plants with a white panicle branch phenotype. In line with the results mentioned above, anatomical structural examination and chlorophyll (Chl) content measurement suggested that wpb1 might play an important role in the regulation of chloroplast development. Further cloning and functional characterization of the wpb1 gene will shed light on the molecular mechanism underlying chloroplast development in rice.


Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Oryza/genética , Fenótipo , Locos de Características Quantitativas , Clorofila/biossíntese , Cloroplastos/metabolismo , Mapeamento Cromossômico , Cromossomos de Plantas/genética , Genes de Plantas/genética , Sequenciamento Completo do Genoma
9.
IEEE Trans Biomed Circuits Syst ; 13(1): 26-37, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30596583

RESUMO

This paper presents a monolithic low power and fast tracking light-to-frequency converter for blood SpO 2 sensing. Normally, the tracking speed and the power consumption are two contradictory characteristics. However, different gain-bandwidth specifications for various ambient light intensities allow the dynamic optimization of the power consumption according to the light intensity. In this paper, the amplifier power consumption is adaptively scaled by the generated light-intensity-positively-correlated control voltage. Thus, the chip total power consumption at low light intensity is significantly decreased. Moreover, the proposed adaptive power scaling is achieved with a continuous analog domain, which does not introduce extra switching noise. The proposed light-to-frequency sensor chip is fabricated by using 0.35  µm CMOS technology with a die area of 1 × 0.9 mm 2. The measurement results show that the pulse light response for any light intensity is no longer than two new output square-wave cycles. The maximum total current consumption is 1.9 mA from a 3.3 V supply voltage, which can be adaptively scaled down to only 0.7 mA if the output frequency is about 25 KHz or lower. The minimum operational supply voltage of the proposed sensor chip is 2.5 V in the temperature range of -25 to 80  °C with 4 KV ESD level (human-body model).


Assuntos
Fontes de Energia Elétrica , Luz , Oxigênio/sangue , Amplificadores Eletrônicos , Simulação por Computador , Humanos , Oximetria , Pulso Arterial , Semicondutores , Temperatura
10.
Clin Exp Pharmacol Physiol ; 46(4): 337-349, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30485484

RESUMO

Parkinson's disease (PD) is a progressive neurodegenerative disorder pathologically characterized by the loss of dopaminergic (DA) neurons in the substantia nigra pars compacta (SNpc). Chronic neuroinflammation is one of the hallmarks of PD pathophysiology. Cathepsin D (CathD), a soluble aspartic protease, has been reported to play an important role in neurodegenerative diseases such as PD. This research focuses on the role of CathD and the molecular mechanisms involved in the process of neuroinflammation and neurotoxicity. We use 1-methyl-4phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP)-challenged mice and lipopolysaccharide (LPS)-induced murine microglia BV2 cells as the in vivo and in vitro models, respectively. The effect of CathD on the neuroinflammation, cytotoxicity and the underlying mechanisms associated with NF-κB signalling pathway are investigated. Data showed that MPTP induces motor deficit, inflammation and depletion of dopaminergic neurons in PD model mice. Notably, cathD was overexpressed in the SNpc of MPTP-induced PD mice and was highly expressing in LPS-stimulated primary microglial cells and BV-2 cells. Furthermore, knockdown of CathD with lentiviral transduction inhibited LPS-induced neuroinflammation through inhibition of NF-κB signalling pathway primarily by regulating the NF-κB p65 nuclear translocation both in BV-2 and primary microglial cells. Additionally, knockdown of CathD protected the activated-microglia induced dopaminergic neurons MN9D cells from neurotoxicity as well as apoptosis. Our findings bring a new insight into understanding the complex mechanisms underlying the pathogenesis of PD and provide a novel target to attenuate the excessive neuroinflammatory responses in the treatment of PD.

11.
Nanoscale Res Lett ; 13(1): 407, 2018 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-30564991

RESUMO

FeOOH nanosheets on porous diatomite have been successfully prepared by a facile two-step hydrothermal approach for supercapacitors, and then α-Fe2O3 and γ-Fe2O3 nanostructures are obtained via calcination under different atmospheres and temperatures. The morphologies and structures of all the samples are investigated in detail to make the hierarchical architecture clear. Besides, systemic tests are carried out in 1 M Na2SO4 electrolyte to characterize the electrochemical properties of these materials. Among the iron-related composite electrodes, diatomite@FeOOH owns the highest specific capacitance (157.9 F g-1 at a current density of 0.5 A g-1) and best cycling performance (98.95% retention after 1000 cycles), which is considered to be a potential material for high-performance supercapacitors. Furthermore, the synthesizing strategy can be extended to the preparation of other metallic oxide-derived functional materials towards energy storage and conversion.

12.
Case Rep Gastrointest Med ; 2018: 3954260, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30425863

RESUMO

Gastric cancer is a malignant tumor with a high degree of malignancy. Multiple liver metastases from gastric cancer (LMGCs) are common. However, the treatment of LMGCs is very difficult. It is rare to achieve complete remission (CR) and long-term survival after treatment. We present the case of a patient with gastric adenocarcinoma and multiple liver metastases who showed CR for more than 33 months after perioperative EOX (epirubicin, oxaliplatin, and capecitabine) combination chemotherapy with radical distal gastrectomy and resection of liver metastases. The patient is still in follow-up without tumor recurrence. These findings suggest that LMGC does not necessarily mean a poor prognosis; preoperative chemotherapy combined with surgery may be a good treatment option for LMGC in selected patients. Further studies are needed to support this treatment approach.

13.
Cell Physiol Biochem ; 44(1): 133-151, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29130967

RESUMO

BACKGROUND/AIMS: Lung cancer (LC) continues to be one of the most prevalent cancers around the world. During this study we aimed to investigate the involvement of endoplasmic reticulum stress (ERS) in autophagy, apoptosis, and chemotherapy resistance of mutant p53 LC cells. METHODS: Immunohistochemistry was employed to help determine the p53 mutation status of cancer cells from 92 primary LC patients, who were subsequently assigned to either the mutant p53 (n = 39) or wild-type p53 group (n = 53). RESULTS: Mutant p53 cells exhibited increased expression of the C/EBP homologous protein (CHOP), glucose-regulated protein 78 (GRP78), and inositol-requiring enzyme-1α (IRE1α). The Mutant p53 cells were also found to be sensitive to chemotherapy and displayed decreased expression of PI3K, Akt, and mTOR. The mutant p53 cell lines were treated with tunicamycin to induce ERS and rapamycin in order to inhibit mTOR. Both agents increased the expression of CHOP, GRP78, IRE1α, LC3-II/LC3-I, Atg5, Atg7, caspase-3, caspase-12, cleaved caspase-3, cleaved caspase-12, as well as decreases in cell proliferation as well as the expression levels of PI3K, Akt, and mTOR. Enhanced levels of cell apoptosis and reduced chemotherapy resistance were also detected. CONCLUSION: The findings of our study suggest that ERS promotes autophagy and apoptosis, while acting to reduce chemotherapy resistance in mutant p53 LC cells by downregulating the PI3K/Akt/mTOR signaling pathway.


Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Estresse do Retículo Endoplasmático , Neoplasias Pulmonares/patologia , Proteína Supressora de Tumor p53/genética , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Feminino , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/metabolismo , Masculino , Pessoa de Meia-Idade , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/efeitos dos fármacos , Serina-Treonina Quinases TOR/genética , Serina-Treonina Quinases TOR/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Tunicamicina/farmacologia , Tunicamicina/uso terapêutico
14.
Oncotarget ; 8(32): 52960-52974, 2017 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-28881786

RESUMO

We investigated the effects of tumor suppressor candidate 3 (TUSC3) on autophagy in human non-small cell lung cancer (NSCLC) cells. A total of 118 NSCLC patients (88 males and 30 females) who underwent surgery at our institute were enrolled in the study. Immunohistochemical analysis revealed that TUSC3 protein expression was lower in NSCLC specimens than adjacent normal tissue. Correspondingly, there was greater methylation of TUSC3 in NSCLC than adjacent normal tissue. After transient transfection of A549 NSCLC cells with constructs designed to up-regulate or down-regulate TUSC3 expression, we analyzed the effects of inhibiting the Wnt pathway (XAV939) and autophagy (chloroquine, CQ) on the behavior of NSCLC cells. We also performed TOP/FOP-Flash reporter assays, MTT assays, Annexin V-FITC/propidium iodide staining, and acridine orange staining to evaluate Wnt/ß-catenin signaling, cell proliferation, apoptosis, and autophagy, respectively. Expression of Wnt/ß-catenin pathway components and autophagy-related proteins was analyzed using qRT-PCR and Western blotting. We found that TUSC3 inhibited cell proliferation and promoted both apoptosis and autophagy in A549 cells. In addition, TUSC3 increased expression of autophagy-related proteins. It also increased expression of Wnt/ß-catenin signaling pathway components and promoted nuclear transfer of ß-catenin, resulting in activation of Wnt/ß-catenin signaling. TUSC3 thus induces autophagy in human NSCLC cells through activation of the Wnt/ß-catenin signaling pathway.

15.
Mol Med Rep ; 16(3): 2785-2790, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28713909

RESUMO

The present study aimed to construct a lentiviral RNA interference (RNAi) vector targeting the transforming growth factor ß1 (TGFß1) gene of rats, in order to examine its effect on silencing of the TGFß1 gene and on the expression of collagen type 1 α1 (Col1a1) in HSC­T6 rat hepatic stellate cells. Three RNAi sites of the TGFß1 gene were selected according to its CDs sequence. Three pairs of small interfering RNA (siRNA) of these RNAi sites were synthesized and then transfected into HSC­T6 cells, respectively, to confirm the optimal siRNA sequence via reverse transcription­polymerase chain reaction analysis. Subsequently, shRNA targeting the sequence of the optimal siRNA was designed, synthesized and annealed to form a double­stranded structure. The annealed oligonucleotide fragment was cloned into pGreenPuro plasmids to establish the pGreenPuro/TGFß1 shRNA lentiviral vector, which was then transfected into 293T cells, following identification by restriction enzyme digestion and sequencing for the production of lentiviral particles exhibiting high reactivity. These particles were used to infect HSC­T6 cells, following which the expression of GFP in the transfected cells was observed under an inverted microscope. The effects on TGFß1 gene silencing and the expression levels of Colla1 were detected at the mRNA and protein levels. The results provided confirmation of the optimal siRNA sequence. Enzyme digestion and sequencing verified successful construction of the pGreenPuro/TGFß1 shRNA lentiviral vector. This lentiviral vector effectively silenced the TGFß1 gene in the HSC­T6 cells, and inhibited the expression of Col1a1 at the mRNA and protein levels. Taken together, the lentiviral RNAi vector targeting the TGFß1 gene of rats was successfully constructed, which effectively silenced the TGFß1 gene of the HSC­T6 cells and inhibited the expression of Col1a1.


Assuntos
Colágeno Tipo I/genética , Células Estreladas do Fígado/metabolismo , Interferência de RNA , Fator de Crescimento Transformador beta1/genética , Animais , Linhagem Celular , Regulação da Expressão Gênica , Lentivirus/genética , RNA Interferente Pequeno/genética , Ratos
16.
Anal Biochem ; 533: 26-33, 2017 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-28610874

RESUMO

Positively-charged nylon membrane (NM) is a general solid-phase support for nucleic acid detection due to its convenient immobilization of nucleic acid materials by direct electrostatic adherence and simple UV crosslinking. Rolling circle amplification (RCA) is a widely used isothermal DNA amplification technique for nucleic acid detection. Near-infrared fluorescence (NIRF) is a new fluorescence technique with high sensitivity due to low background. This study developed a simple method for detecting nucleic acid molecules by combining the advantages of NM, RCA and NIRF, named NIRF-based solid phase RCA on nylon membrane (NM-NIRF-sRCA). The detection system of this method only need two kinds of nucleic acid molecules: target-specific probes with a RCA primer (P) at their 3' end and a rolling circle (RC). The detection procedure consists of four steps: (1) immobilizing detected nucleic acids on NM by UV crosslinking; (2) hybridizing NM with specific probes and RC; (3) amplifying by a RCA reaction containing biotin-dUTP; (4) incubating NM with NIRF-labeled streptavidin and imaging with a NIRF imager. The method was fully testified by detecting oligonucleotides, L1 fragments of various HPV subtypes cloned in plasmid, and E.coli genomic DNA. This study thus provides a new facile method for detecting nucleic acid molecules.


Assuntos
Técnicas Biossensoriais , Técnicas de Amplificação de Ácido Nucleico , Ácidos Nucleicos/isolamento & purificação , Biotina/química , Primers do DNA/genética , Humanos , Membranas/química , Hibridização de Ácido Nucleico , Ácidos Nucleicos/genética , Nylons/química , Estreptavidina/química
17.
Parasite ; 24: 7, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28224883

RESUMO

Toxoplasma gondii is a zoonotic protozoan parasite that infects a wide range of warm-blooded animals throughout the world. In the present study, antibodies to T. gondii were determined using a commercial indirect hemagglutination (IHA) test in wild animals in a zoo. Three of 11 giraffes (Giraffa camelopardalis) (27%), 1 of 5 wolves (Canis lupus laniger) (20%), 1 of 6 hippopotamuses (Hippopotamus amphibious) (17%), and 2 of 9 tundra swans (Cygnus columbianus) (22%) were found to be positive. No antibodies were detected in leopards (Panthera pardus), wild geese (Anser cygnoides), and Eastern grey kangaroos (Macropus giganteus). Domestic species from 13 counties of Jiangxi Province, China were also investigated by an indirect hemagglutination (IHA) test. Thirty-five of 340 goats (10%), 94 of 560 water buffaloes (17%), and 4 of 35 cattle (11%) were found to be seropositive. This is the first report of T. gondii infection in animals kept in zoos and domestic animals in this province.


Assuntos
Animais Domésticos/parasitologia , Animais de Zoológico/parasitologia , Anticorpos Antiprotozoários/sangue , Toxoplasma/imunologia , Toxoplasmose Animal/epidemiologia , Animais , Anseriformes/parasitologia , Artiodáctilos/parasitologia , Doenças das Aves/epidemiologia , Doenças das Aves/parasitologia , Búfalos/parasitologia , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/parasitologia , China/epidemiologia , Gansos/parasitologia , Girafas/parasitologia , Doenças das Cabras/epidemiologia , Doenças das Cabras/parasitologia , Cabras , Macropodidae/parasitologia , Panthera/parasitologia , Estudos Soroepidemiológicos , Lobos/parasitologia
19.
Chin J Integr Med ; 23(1): 70-75, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27679442

RESUMO

OBJECTIVE: To investigate the effect of Shaoyao Gancao Decoction (, SGD) on the pharmacokinetics of intravenously administered paclitaxel in rats. METHODS: Paclitaxel was intravenously administered to rats (3 mg/kg) with or without the concomitant administration of SGD (752 mg/kg, a single day or 14 consecutive days pretreatment). The paclitaxel in the serum was quantified using a simple and rapid ultra performance liquid chromatography (UPLC) method for the pharmacokinetic study. The pharmacokinetic parameters were calculated via a non-compartment model using the computer program DAS 2.0. RESULTS: The pharmacokinetic parameters of paclitaxel were significantly altered in response to 14 consecutive days of pretreatment with SGD. The area under the curve (AUC0-t, from 4 820±197 to 4 205±186 ng·mL-1·-1) and AUC0-∞ (from 5 237±280 to 4 514±210 ng·mL-1·-1) significantly decreased in response to the 14-day pretreatment with SGD. The values of Vdss (L/kg) were 10.74±1.08 and 9.35±0.49, those of CL (L/kg) were 0.67±0.03 and 0.57±0.03 and the t1/2 (h) values were 11.17±0.84 and 11.32±0.93, respectively, for the 14-day SGD pretreatment and intravenous paclitaxel alone. The AUC0-t and AUC0-∞ values decreased by 13% and 14% (P<0.01), respectively. The area under the curve decreased signifificantly (P<0.01), and the total clearance increased by 1.2-fold (P<0.01), after 14 consecutive days of pretreatment with SGD. A single-day pretreatment with SGD did not signifificantly affect the pharmacokinetic parameters of paclitaxel. CONCLUSIONS: SGD administration for 14 consecutive days increased the metabolism of paclitaxel, while a 1-day pretreatment had little effect. The results would contribute important information to the study on interaction between Chinese medicines and chemotherapy and also help to utilize SGD better in the adjunctive therapy of cancer patients.


Assuntos
Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/uso terapêutico , Paclitaxel/administração & dosagem , Paclitaxel/farmacocinética , Animais , Cromatografia Líquida de Alta Pressão , Injeções Intravenosas , Masculino , Paclitaxel/sangue , Paclitaxel/química , Ratos Sprague-Dawley , Padrões de Referência , Fatores de Tempo
20.
Sci Rep ; 6: 39303, 2016 12 14.
Artigo em Inglês | MEDLINE | ID: mdl-27966632

RESUMO

Surfactin, a natural lipopeptide, can be used both as parenteral and non-parenteral adjuvant for eliciting immune response. However, the mechanisms that confer its adjuvant properties have not been fully explored. By staining with NHS-Rhodamine B labeled surfactin and Mito-Tracker Green, we found surfactin could penetrate into macrophages to bind with mitochondria, following induce ROS that could be inhibited by mitochondria-dependent ROS inhibitor. ROS enhanced p38 MAPK and JNK expression, as well their phorsphorylation, following activated NF-κB nuclear translocation in macrophages that was obviously inhibited by mitochondria-dependent ROS inhibitor. However, inhibition of ROS production only weakened p38 MAPK and JNK expression, but not their phosphorylation in macrophages. As a result, surfaction could activate NF-κB to release TNF-α by the mitochondria-dependent ROS signalling pathway. ROS also induced macrophages apoptosis to release endogenous danger signals, following activated inflammasomes of NLRP1, NLRP3, IPAF and AIM2 in vitro and only NLRP1 in vivo, as well caspase-1 and IL-1 in macrophages, which were significantly inhibited by pre-treatment with ROS inhibitors. Collectively, surfactin as a kind of non-pathogen-associated molecular patterns, modulates host innate immunity by multiple signalling pathways, including induction of mitochondria-dependent ROS, activating MAPKs and NF-κB, and inducing cell apoptosis to realease endogenous danger signals for activation of inflammasomes.


Assuntos
Inflamassomos/metabolismo , Lipopeptídeos/metabolismo , Macrófagos/imunologia , Mitocôndrias/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Peptídeos Cíclicos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Adjuvantes Imunológicos/metabolismo , Animais , Macrófagos/efeitos dos fármacos , Camundongos , Mitocôndrias/efeitos dos fármacos , Células RAW 264.7
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