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1.
J Med Microbiol ; 70(8)2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34397349

RESUMO

Introduction. Lactococcus petauri LZys1 (L. petauri LZys1) is a type of lactic acid bacteria (LAB), which was initially isolated from healthy human gut.Hypothesis/Gap Statement. It was previously anticipated that L. petauri LZys1 has potential characteristics of probiotic properties. The genetic structure and the regulation functions of L. petauri LZys1 need to be better revealed.Aim. The aim of this study was to detect the probiotic properties L. petauri LZys1 and to reveal the genome information related to its genetic adaptation and probiotic profiles.Methodology. Multiple in vitro experiments were carried out to evaluate its lactic acid-producing ability, resistance to pathogenic bacterial strains, auto-aggregation and co-aggregation ability, and so on. Additionally, complete genome sequencing, gene annotation, and probiotic associated gene analysis were performed.Results. The complete genome of L. petauri LZys1 comprised of 1 985 765 bp, with a DNA G+C content of 38.07 %, containing 50 tRNA, seven rRNA, and four sRNA. A total of 1931 genes were classified into six functional categories by Kyoto Encyclopaedia of Genes and Genomes (KEGG) database. The neighbour-joining phylogeny tree based on the whole genome of L. petauri LZys1 and other probiotics demonstrated that L. petauri LZys1 has a significant similarity to Lactococcus garvieae. The functional genes were detected to expound the molecular mechanism and biochemical processes of its potential probiotic properties, such as atpB gene.Conclusion. All the results described in this study, together with relevant information previously reported, made L. prtauri LZys1 a very interesting potential strain to be considered as a prominent candidate for probiotic use.


Assuntos
Trato Gastrointestinal/microbiologia , Genoma Bacteriano , Lactococcus , Probióticos , Animais , Bactérias/crescimento & desenvolvimento , Bactérias/patogenicidade , Sequência de Bases , Fezes/microbiologia , Genes Bacterianos , Humanos , Lactococcus/citologia , Lactococcus/genética , Lactococcus/isolamento & purificação , Lactococcus/fisiologia , Masculino , Anotação de Sequência Molecular , Mariposas/microbiologia , Filogenia , Polissacarídeos Bacterianos/biossíntese , Polissacarídeos Bacterianos/genética , Sequenciamento Completo do Genoma , Adulto Jovem
2.
Mol Med Rep ; 19(5): 3955, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30896791

RESUMO

The authors wish to retract their article entitled "Generation and characterization of induced pluripotent stem cells from guinea pig fetal fibroblasts" published in Molecular Medicine Reports 15, 3690­3698, 2017. Following the publication of this article, the authors noted that the description of the reprogramming method and cell cultured conditions in the article was inconsistent with the experimental facts, due to their oversight and personnel issues in terms of who performed the experimental work and who wrote up the paper. Some of the experimental approaches were wrongly described in the Materials and methods section. Namely, the feeder cells which were used in this reprogramming were mouse embryonic fibroblasts, not guinea fibroblast cells, as was reported; the reprogramming medium contained 15% knockout serum replacement; the culture medium contained 10% ESC qualified fetal bovine serum and 10% knockout serum replacement; and 293T cells were transfected using the Quick Shuttle­293 system. Due to the authors' oversight, Figs 2A and C, and Fig. 3 on p. 3,694 do not match with the textual description. In view of these serious errors, which the authors are not able to rectify, they have requested that this article be retracted from the publication. All the named authors agree to this retraction. The authors regret any inconvenience to the readers that this retraction will cause.

3.
Int J Mol Med ; 42(1): 131-140, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29620145

RESUMO

Leukemia inhibitory factor (LIF) is the most pleiotropic cytokine of the interleukin­6 family, and is widely used to establish and maintain pluripotent stem cells, particularly mouse pluripotent stem cells. However, no reports have fully elucidated the application of LIF in marmoset induced pluripotent stem cell (iPSC) culture, particularly the underlying mechanisms. To demonstrate the feasibility of the application of LIF to marmoset iPSCs, the present study assessed these cells in the presence of LIF. Cell proliferation was measured using MTT assay, cell apoptosis was determined by flow cytometric analysis of fluorescein isothiocyanate Annexin V staining and the differentially expressed genes were analysed using Digital Gene Expression (DGE) analysis. The altered expression of pluripotency­associated genes was confirmed by reverse transcription­quantitative polymerase chain reaction and western blot analysis. Furthermore, following treatment with LY294002, cell proliferation was measured by MTT assay and protein levels were confirmed by western blot analysis. The results showed that LIF significantly promoted the number of proliferating cells, but had no effect on apoptosis. Digital Gene Expression analysis was used to examine the differentially expressed genes of marmoset iPSCs in the presence of LIF. The results showed that the pluripotency­associated transcription factor­encoding gene T­box 3 (Tbx­3) was activated by LIF. Notably, LIF increased the levels of phosphorylated (p­)AKT and Tbx­3 in the marmoset iPSCs. Furthermore, pretreatment with LY294002, an inhibitor of phosphoinositide 3­kinase (PI3K), significantly impaired the LIF­induced upregulation of p­AKT and Tbx­3 in the marmoset iPSCs, suggesting that the PI3K/Akt signaling pathway is involved in this regulation. Taken together, the results suggested that LIF is effective in maintaining marmoset iPSCs in cultures, which is associated with the activation of Tbx­3 through regulation of the PI3K/Akt signaling pathway.


Assuntos
Células-Tronco Pluripotentes Induzidas/citologia , Células-Tronco Pluripotentes Induzidas/enzimologia , Fator Inibidor de Leucemia/farmacologia , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Proteínas com Domínio T/metabolismo , Animais , Apoptose/efeitos dos fármacos , Callithrix , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/efeitos dos fármacos , Células-Tronco Pluripotentes Induzidas/efeitos dos fármacos , Camundongos , Modelos Biológicos , Transdução de Sinais/efeitos dos fármacos , Proteínas com Domínio T/genética
4.
Stem Cell Res Ther ; 9(1): 49, 2018 02 26.
Artigo em Inglês | MEDLINE | ID: mdl-29482598

RESUMO

BACKGROUND: Induced pluripotent stem cells (iPS) represent an innovative source for the standardized in vitro generation of macrophages (Mφ). Mφ show great promise in disease pathogenesis, particularly tuberculosis. However, there is no information about human iPS-derived (hiPS) macrophages (hiPS-Mφ) in response to tuberculosis infection. METHODS: In the present study, macrophages derived from hiPS were established via embryoid body (EB) formation by using feeder-free culture conditions, and the human monocyte cell line THP-1 (THP-1-Mφ) was used as control. iPS-Mφ were characterized by using morphology, Giemsa staining, nonspecific esterase staining (α-NAE), phagocytosis, and surface phenotype. Additionally, after treatment with Bacillus Calmette-Guérin (BCG) for 24 h, cell apoptosis was detected by using an Annexin V-FITC Apoptosis Detection assay. The production of nitric oxide (NO), expression of tumor necrosis factor alpha (TNF-α), activity of apoptosis-related protein cysteine-3 (Caspase-3) and expression of B-cell lymphoma-2 (Bcl-2) were analyzed. RESULTS: With respect to morphology, surface phenotype, and function, the iPS-Mφ closely resembled their counterparts generated in vitro from a human monocyte cell line. iPS-Mφ exhibited the typically morphological characteristics of macrophages, such as round, oval, fusiform and irregular characteristics. The cells were Giemsa-stained-positive, α-NAE-positive, and possessed phagocytic ability. iPS-Mφ express high levels of CD14, CD11b, CD40, CD68, and major histocompatibility complex II (MHC-II). Moreover, with regard to the apoptotic rate, the production of NO, expression of TNF-α, and activity of Caspase-3 and Bcl-2, iPS-Mφ closely resemble that of their counterparts generated in vitro from human monocyte cell line in response to BCG infection. The rate of apoptosis of BCG-treated iPS-Mφ was 37.77 ± 7.94% compared to that of the untreated group at 4.97 ± 1.60% (P < 0.01) by using Annexin V-FITC Apoptosis Detection. Additionally, the rate of apoptosis of BCG-treated THP-1-Mφ was 37.1 ± 2.84% compared to that of the untreated group at 6.19 ± 1.68% (P < 0.001). The expression of TNF-α and the production of NO were significantly increased (P < 0.001), and the activity of Caspase-3 was increased. However, the expression of Bcl-2 was inhibited (P < 0.001). CONCLUSIONS: Our results demonstrate that Mφ derived from hiPS perform the immunological function in response to Bacillus Calmette-Guérin infection by undergoing apoptosis, increasing the production of NO and expression of TNF-α. Thus, our study may help to overcome the limitations of research into certain rare diseases due to the lack of adequate supply of disease-specific primary cells.


Assuntos
Apoptose/imunologia , Regulação da Expressão Gênica/imunologia , Células-Tronco Pluripotentes Induzidas/imunologia , Macrófagos/imunologia , Mycobacterium bovis/imunologia , Óxido Nítrico/imunologia , Tuberculose/imunologia , Fator de Necrose Tumoral alfa/imunologia , Humanos , Células-Tronco Pluripotentes Induzidas/microbiologia , Células-Tronco Pluripotentes Induzidas/patologia , Macrófagos/microbiologia , Macrófagos/patologia , Células THP-1 , Tuberculose/microbiologia , Tuberculose/patologia
5.
Mol Med Rep ; 15(6): 3690-3698, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28393187

RESUMO

Induced pluripotent stem cells (iPS) represent an important tool to develop disease­modeling assays, drug testing assays and cell­based replacement therapies. The application of iPS in these fields requires the development of suitable animal models. Of the suitable species, guinea pigs are particularly important and offer significant advantages. Successful iPS generation has been accomplished in a number of species; however, it has not been reported in the guinea pig. The present study successfully generated iPS from guinea pigs (giPS) using single polycistronic virus transduction with mouse octamer­binding transcription factor 4 (Oct4), sex determining region Y­box 2 (Sox2), Kruppel­like factor 4 and c­Myc. The giPS cell lines were cultured in media containing leukemia inhibitory factor and guinea pig fibroblast cells were used as feeder cells. These cultures were expanded under feeder­free culture conditions using ESGRO Complete Plus Clonal Grade medium containing 15% fetal bovine serum on gelatin­coated dishes. The resultant cells had a normal karyotype, exhibited alkaline phosphatase activity and expressed the pluripotency markers Oct4, Sox2 and Nanog. The cells differentiated in vivo to form teratomas that contained all three germ layers of the tissue cells. The generation of giPS may facilitate future studies investigating the mechanisms underlying innate immunity, particularly for tuberculosis. These experiments provide proof of principle that iPS technology may be adapted to use the guinea pig as a model of human diseases.


Assuntos
Reprogramação Celular/genética , Fibroblastos/citologia , Fibroblastos/metabolismo , Células-Tronco Pluripotentes Induzidas/citologia , Células-Tronco Pluripotentes Induzidas/metabolismo , Animais , Biomarcadores , Técnicas de Cultura de Células , Linhagem Celular , Transformação Celular Neoplásica , Feminino , Cobaias , Cariotipagem , Camundongos , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
6.
J Matern Fetal Neonatal Med ; 28(10): 1165-9, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25053194

RESUMO

OBJECTIVE: To investigate the mRNA and protein expression of FK506-binding protein 52 (FKBP52) in the chorionic villi of patients with recurrent spontaneous abortion (RSA) and normal women during early pregnancy. METHODS: Fresh chorionic villus tissues were collected from 60 subjects. A total of 30 patients with a history of RSA were enrolled into the RSA group and 30 normal pregnant women were enrolled into the control group. The FKBP52 mRNA expression levels in chorionic villi of the RSA patients and healthy controls were measured via semiquantitative RT-PCR. The protein distribution and expression levels of FKBP52 in chorionic villi were analyzed through immunohistochemistry (IHC). The correlation between FKBP52 expression and RSA was analyzed. RESULTS: We demonstrated that FKBP52 mRNA is expressed in chorionic villi samples of normal pregnancy and RSA. RSA patients exhibited significantly lower FKBP52 gene expression levels compared with those in normal pregnancies (p < 0.05). FKBP52 immunoreactivity in chorionic villi was mainly observed in trophoblast cell cytoplasm. The FKBP52 protein expression levels in the chorionic villi of RSA patients was significantly lower than in normal women during pregnancy (p < 0.05). CONCLUSIONS: FKBP52 protein levels were decreased in the chorionic villi of RSA patients, which indicate that the decrease in FKBP52 may be associated with RSA. The low FKBP52 mRNA expression level, which is consistent with the IHC result, may affect embryonic development and even lead to abortion. FKBP52 may be involved in the pathogenesis of RSA and new therapies that increase the FKBP52 expression may help treat RSA.


Assuntos
Aborto Habitual/metabolismo , Vilosidades Coriônicas/metabolismo , Proteínas de Ligação a Tacrolimo/genética , Proteínas de Ligação a Tacrolimo/metabolismo , Adulto , Feminino , Humanos , Imuno-Histoquímica , Gravidez , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa
7.
PLoS One ; 8(9): e75759, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24098721

RESUMO

Recurrent spontaneous abortion (RSA) is a health problem that affects approximately 1% to 5% reproductive age woman. Yet, in around half of these patients, the mechanism for RSA is unexplained. Recent studies have indicated that placental ischemia/hypoxia and endothelial dysfunction are important factors in miscarriage. Other studies have indicated that the level and expression of soluble FMS-like tyrosine kinase-1 (sFlt1) is increased under a hypoxic environment. However, decreased sFlt-1 in the maternal circulation during the first trimester has recently been proposed as a potential marker for identifying risk of pregnancy loss. In this prospective study clinical samples were obtained within a short time after the fetal death, protein expression and maternal serum levels of sFlt1 were assessed and compared to samples taken from those with normal pregnancies. Our results indicate that levels of VEGF and sFlt-1 are both increased in women during early pregnancy compared women that are not pregnant (p<0.05) indicating that VEGF and sFlt-1 are both associated with pregnancy. More importantly, we detected a significant (p<0.05) increase in sFlt1 and VEGF levels and expression in the RSA patients who suffered subsequent miscarriages compare to controls. These results demonstrate that there is likely a relationship between VEGF, sFlt-1 and RSA suggesting that the high levels and over expression of sFlt-1 and VEGF might be associated with the pathogenesis of RSA.


Assuntos
Aborto Habitual/etiologia , Aborto Habitual/fisiopatologia , Fator A de Crescimento do Endotélio Vascular/sangue , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/sangue , Estudos de Casos e Controles , China , Feminino , Humanos , Imuno-Histoquímica , Gravidez , Estudos Prospectivos
8.
Wei Sheng Yan Jiu ; 35(3): 345-7, 2006 May.
Artigo em Chinês | MEDLINE | ID: mdl-16921765

RESUMO

OBJECTIVE: To study the correlation of rare earth elements (REEs) in the rats' hair, blood and organs. METHODS: Based on the level of animal weights, 50 healthy male SD rats were randomly divided into five groups including control group and four Citrate REEs level groups (low, middle, high-I and high-II). Before the experiment, the hair of rats' back was eliminated. After the rats were fed for four weeks, the fresh hair of the rats was collected. Except the high-II group, the blood and organs of the others were collected. The high-II group was fed for other four weeks without Citrate REEs. At the end of eighth week, hair, blood and organs of the high-II group were collected. Determination of light REEs concentration in the rats' hair, blood, liver, spleen and bone by ICP-MS. RESULTS: The correlation coefficients of REEs concentration between hair and organs (such as liver, spleen and bone) were more than 0.5, while those of REEs concentration between blood and organs (such as liver spleen and bond) were less than 0.5. In the group H- Il, the Rees concentration in blood, hair, liver, spleen and bone were all decrease, and the REEs concentration of blood was close to that of the control groups. CONCLUSION: The concentrations of REEs in these organs were different. And hair was better than blood as a biomarker to reflect body exposure of REEs.


Assuntos
Exposição Ambiental/análise , Cabelo/química , Fígado/química , Metais Terras Raras/farmacocinética , Animais , Masculino , Metais Terras Raras/análise , Metais Terras Raras/sangue , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Baço/química , Distribuição Tecidual
9.
Zhongguo Zhong Yao Za Zhi ; 29(8): 753-9, 2004 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-15506286

RESUMO

OBJECTIVE: To make an attempt at the multi-element speciation in the Chinese medicinal herbs by determining the concentrations of 25 elements in different extraction solutions. METHOD: Firstly, five Chinese medicinal herbs (Buddleja officinalis, Dictamnus dasycarpus, Myristica fragrans, Albizia judibrissin and Inula japonica) from the same region of China were treated to obtain water-soluble phase, lipid-soluble phase and non-soluble phase by water extraction, organic solvent extraction and acid digestion, respectively. Secondly, Phytolacca acinosa, a Chinese medicinal herb collected from 9 regions of China, was extracted by 0% EtOH, 50% EtOH, 75% EtOH, 95% EtOH, respectively, referring the Chinese Pharmacopoeia. Finally, the concentrations of 25 elements, such as Be, Cr, Cu, Zn, Ge, Sr, Y, Mo, Cd, Tl, Pb and REEs, in the above three phases were determined by ICP-MS. RESULT: Under the optimal conditions, all the 25 elements could be determined with detection limits ranged from 0.003 to 0.71 ng x g(-1). The average recoveries of the elements in P. acinosa were 88% approximately 119%, with the relative standard deviations 1.7% approximately 13.3%. It was observed that the determined 25 elements distributed in all the water-soluble, lipid-soluble and non-soluble phases, indicating that the inorganic species, organicspecies, as well as the protein bound species were coexisted in the herbs. Big differences of the element extraction rates could be found by using different ethanol solutions. CONCLUSION: With the aid of the obtained results, we may increase the extraction of necessary elements while decrease that of the toxic elements from the herbs by choosing a suitable solvent during the drug production.


Assuntos
Medicamentos de Ervas Chinesas/química , Metais Pesados/análise , Phytolacca/química , Plantas Medicinais/química , Oligoelementos/análise , Buddleja/química , Cádmio/análise , Cobre/análise , Dictamnus/química , Medicamentos de Ervas Chinesas/isolamento & purificação , Ecossistema , Chumbo/análise , Molibdênio/análise , Myristica/química , Solventes/química , Zinco/análise
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