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1.
Reprod Sci ; 2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-32046391

RESUMO

In situ production and metabolism of all-trans retinoic acid (RA) in decidual tissue are critically important for endometrial stromal differentiation, embryo implantation, and healthy placentation. However, the cellular source(s) of RA in this tissue has yet to be determined. To identify the primary RA-producing cells in human term decidua, we isolated cells from decidua basalis of delivered placenta and quantified cellular retinal dehydrogenase (RALDH) activity, a major biosynthetic enzyme whose activity determines the synthesis of RA from retinol, using an Aldefluor assay and flow cytometry. RA production in decidual tissue and sorted cell subpopulations was evaluated by liquid chromatography-tandem mass spectrometry. CD14+ cells (macrophages/monocytes) showed > 4-fold higher RALDH activity than stromal cells (CD10+), T cells (CD3+), or non-T lymphocytes (CD3-negative). CD11c+ cells that did not co-express CD14 showed about one-third the RALDH activity of their CD14 co-expressing counterparts. The highest RALDH activity was found in "alternatively activated" M2 macrophages delineated by the simultaneous expression of CD14 and CD163. The greater RA synthesizing capacity of M2 versus CD14+CD163-ve (M1) cells was confirmed by direct quantitation of RA biosynthesis from retinol. RA levels in whole decidua were correlated with M2 cell density but not with stromal cell (CD10+) number, the major cell type comprising the decidua. These results identified M2 monocyte/macrophages as the primary source of RA in human term decidua. This finding may have implications for certain pregnancy complications that are known to be associated with reduced numbers of decidual M2 cells.

2.
J Neuroinflammation ; 16(1): 241, 2019 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-31779628

RESUMO

BACKGROUND: Sepsis-associated encephalopathy (SAE), a diffuse cerebral dysfunction in the absence of direct CNS infection, is associated with increased rates of mortality and morbidity in patients with sepsis. Increased cytokine production and disruption of the blood-brain barrier (BBB) are implicated in the pathogenesis of SAE. The induction of pro-inflammatory mediators is driven, in part, by activation of NF-κΒ. Lipopolysaccharide (LPS), an endotoxin produced by gram-negative bacteria, potently activates NF-κΒ and its downstream targets, including cyclooxygenase-2 (Cox-2). Cox-2 catalyzes prostaglandin synthesis and in the brain prostaglandin, E2 is capable of inducing endothelial permeability. Depletion of polymerase δ-interacting protein 2 (Poldip2) has previously been reported to attenuate BBB disruption, possibly via regulation of NF-κΒ, in response to ischemic stroke. Here we investigated Poldip2 as a novel regulator of NF-κΒ/cyclooxygenase-2 signaling in an LPS model of SAE. METHODS: Intraperitoneal injections of LPS (18 mg/kg) were used to induce BBB disruption in Poldip2+/+ and Poldip2+/- mice. Changes in cerebral vascular permeability and the effect of meloxicam, a selective Cox-2 inhibitor, were assessed by Evans blue dye extravasation. Cerebral cortices of Poldip2+/+ and Poldip2+/- mice were further evaluated by immunoblotting and ELISA. To investigate the role of endothelial Poldip2, immunofluorescence microscopy and immunoblotting were performed to study the effect of siPoldip2 on LPS-mediated NF-κΒ subunit p65 translocation and Cox-2 induction in rat brain microvascular endothelial cells. Finally, FITC-dextran transwell assay was used to assess the effect of siPoldip2 on LPS-induced endothelial permeability. RESULTS: Heterozygous deletion of Poldip2 conferred protection against LPS-induced BBB permeability. Alterations in Poldip2+/+ BBB integrity were preceded by induction of Poldip2, p65, and Cox-2, which was not observed in Poldip2+/- mice. Consistent with these findings, prostaglandin E2 levels were significantly elevated in Poldip2+/+ cerebral cortices compared to Poldip2+/- cortices. Treatment with meloxicam attenuated LPS-induced BBB permeability in Poldip2+/+ mice, while having no significant effect in Poldip2+/- mice. Moreover, silencing of Poldip2 in vitro blocked LPS-induced p65 nuclear translocation, Cox-2 expression, and endothelial permeability. CONCLUSIONS: These data suggest Poldip2 mediates LPS-induced BBB disruption by regulating NF-κΒ subunit p65 activation and Cox-2 and prostaglandin E2 induction. Consequently, targeted inhibition of Poldip2 may provide clinical benefit in the prevention of sepsis-induced BBB disruption.

3.
mSystems ; 4(5)2019 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-31594827

RESUMO

Gut microbiota play important roles in host metabolism, especially in diabetes. However, why different diets lead to similar diabetic states despite being associated with different microbiota is not clear. Mice were fed two high-energy diets (HED) with the same energy density but different fat-to-sugar ratios to determine the associations between the microbiota and early-stage metabolic syndrome. The two diets resulted in different microbiota but similar diabetic states. Interestingly, the microbial gene profiles were not significantly different, and many common metabolites were identified, including l-aspartic acid, cholestan-3-ol (5ß, 3α), and campesterol, which have been associated with lipogenesis and inflammation. Our study suggests that different metabolic-syndrome-inducing diets may result in different microbiota but similar microbiomes and metabolomes. This suggests that the metagenome and metabolome are crucial for the prognosis and pathogenesis of obesity and metabolic syndrome.IMPORTANCE Various types of diet can lead to type 2 diabetes. The gut microbiota in type 2 diabetic patients are also different. So, two questions arise: whether there are any commonalities between gut microbiota induced by different pro-obese diets and whether these commonalities lead to disease. Here we found that high-energy diets with two different fat-to-sugar ratios can both cause obesity and prediabetes but enrich different gut microbiota. Still, these different gut microbiota have similar genetic and metabolite compositions. The microbial metabolites in common between the diets modulate lipid accumulation and macrophage inflammation in vivo and in vitro This work suggests that studies that only use 16S rRNA amplicon sequencing to determine how the microbes respond to diet and associate with diabetic state are missing vital information.

4.
J Cell Biochem ; 120(12): 19457-19468, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31478245

RESUMO

Gastric cancer (GC) is one of the most malignant tumors that seriously threaten human health. Increased reports have indicated that long noncoding RNAs (lncRNAs) are associated with GC. This study aims to investigate the regulatory role of colon cancer-associated transcript-1 (CCAT1) in GC. The results exhibited the fact that CCAT1 was expressed higher in 57 GC tissue samples than in 57 paired adjacent normal tissue samples. The expression of CCAT1 was also increased in GC cell lines (MKN45, Hs746T, and SGC-7901) compared with the gastric epithelial cell line GES-1. Besides this, decreased cell proliferation with increased cell apoptosis was detected in SGC-7902 cells transfected with CCAT1 short hairpin RNA (shRNA). At the same time, a lower cell invasion ability was measured in SCG-7901 cells transfected with CCAT1 shRNA.In addition, miR-219-1 was predicted and convinced a direct target of CCAT1. The expression of miR-219-1 was decreased in GC tissues and GC cell lines. Further studies demonstrated that the roles of CCAT1 in cell proliferation, apoptosis, and invasion were inhibited by miR-219-1. Finally, in vivo experiment indicated that tumor growth of GC was suppressed through knockdown of CCAT1. In conclusion, these results suggested that CAT1 promotes the tumorigenesis and progression of GC by negatively regulating miR-219-1.

5.
Oncol Lett ; 16(3): 3489-3498, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30127953

RESUMO

Gastric cancer can be a fatal tumor and therefore represents one of the primary challenges in modern oncology. Survivin and X-linked inhibitor of apoptosis protein (XIAP) are members of the IAP family, which exerts a strong inhibitory effect on cellular apoptosis. In previous studies, the expression levels of survivin and XIAP have been demonstrated to influence the prognosis of patients with gastric cancer; therefore, the present study investigated the effect of silencing survivin and XIAP on the biological activity of the gastric cancer HGC-27 cell line. It was demonstrated that the expression levels of survivin and XIAP were significantly increased in gastric cancer tissues, compared with the adjacent non-tumor tissues. Furthermore, it was observed that the expression levels of survivin and XIAP were similarly elevated in gastric cancer HGC-27 cells, compared with normal gastric epithelial GES-1cells. Furthermore, small interfering RNA-mediated surviving- or XIAP-knockdown, in addition to the dual knockdown of survivin and XIAP, inhibited the proliferation and promoted the apoptosis of HGC-27 cells. Simultaneous inhibition of XIAP and survivin expression was more effective, compared with inhibition of XIAP or survivin alone. These results indicated that the dual knockdown of survivin and XIAP may be an effective strategy for treating gastric cancer in the future.

6.
J Cell Biochem ; 2017 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-29231252

RESUMO

Gastric cancer (GC) is one of the most malignant tumors that seriously threaten to human health. Increased reports indicated that long non-coding RNAs (lncRNAs) were associated with GC. This study aims to investigate the regulatory role of colon cancer associated transcript-1 (CCAT1) in GC. The results exhibited that CCAT1 was higher expressed in 57 GC tissue samples than in 57 paired adjacent normal tissue samples. The expression of CCAT1 was also increased in GC cell lines (MKN45, Hs746T and SGC-7901) compared with gastric epithelial cell line GES-1. Besides, decreased cell proliferation with increased cell apoptosis were detected in SGC-7902 cells transfected with CCAT1 shRNA. At the same time, lower cell invasion ability was measured in SCG-7901 cells transfected with CCAT1-shRNA. In addition, miR-219-1 was predicted and convinced a direct target of CCAT1. The expression of miR-219-1 was declined in GC tissues and GC cell lines. Further studies demonstrated that the roles of CCAT1 on cell proliferation, apoptosis and invasion were inhibited by miR-219-1. At last, the in vivo experiment indicated that tumor growth of GC was suppressed through knockdown of CCAT1. In conclusion, these results suggested that CAT1 promotes the tumorigenesis and progression of GC by negative-regulating miR-219-1. This article is protected by copyright. All rights reserved.

7.
Oncol Rep ; 38(6): 3465-3472, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29039602

RESUMO

Gastric cancer (GC) is one of the most common malignant diseases worldwide. Although significant progress has been made in the early detection and treatment of GC over the past decades, the prognosis is still not satisfactory and the underlying mechanisms of carcinogenesis remain unknown. Long non-coding RNA MIAT has been established as a key player in the regulation of various biological and pathological processes including chronic lymphocytic leukemias, acute myocardial infarction and neuroendocrine prostate cancer. However, the function of MIAT in GC remains largely unknown. The expressions of lncRNA MIAT, miR-29a-3p and HDAC4 mRNA were analysed using quantitative real-time PCR (qRT-PCR). RNA interference approach was used to investigate the cellular functions of MIAT and miR-29a-3p. Cell Counting Kit-8 (CCK-8) assay and flow cytometry assay were performed to detect cell proliferation and apoptosis. Cell migration and invasion abilities were evaluated by Transwell assays. In the present study, we first confirmed the high expression level of MIAT in GC tissues and cell lines. In addition, knockdown of MIAT suppressed the proliferation, migration and invasion of GC cells in vitro. Furthermore, our results demonstrated that MIAT competitively binds to miR-29a-3p and consequently upregulates the expression of HDAC4, which is a downstream target of miR-29a-3p. In conclusion, the present study highlighted the involvement of the MIAT/miR-29a-3p/HDAC4 axis in the development of GC, which provided potential diagnostic and therapeutic targets for GC.


Assuntos
Histona Desacetilases/genética , MicroRNAs/genética , RNA Longo não Codificante/genética , Proteínas Repressoras/genética , Neoplasias Gástricas/genética , Idoso , Apoptose/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Pessoa de Meia-Idade , Prognóstico , RNA Mensageiro/genética , RNA Interferente Pequeno/genética , Neoplasias Gástricas/patologia
8.
Acupunct Med ; 35(3): 216-223, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27852563

RESUMO

BACKGROUND: Electroacupuncture (EA) may have a role in the treatment of diarrhoea symptoms. Serotonin (5-hydroxytryptamine, 5-HT) is an important neurotransmitter and paracrine signalling molecule in the gastrointestinal (GI) tract, which initiates peristaltic, secretory, vasodilatory, vagal and nociceptive reflexes. In addition, according to the results of our previous report, EA stimulation mediates GI peristalsis by increasing expression of 5-HT and tryptophan hydroxylase (TPH). AIM: To investigate the effect of EA at acupuncture points ST25 and BL25 in a rat model of diarrhoea. METHODS: A diarrhoea-predominant irritable bowel syndrome (IBS-D) model was induced by Folium Sennae in 24 rats, which remained untreated (n=6) or received EA at ST25 (n=6), BL25 (n=6) or the combination of ST25 and BL25 (n=6). A control group of healthy rats was also included (n=6). After treatment, changes in loose stool and small intestine transit rates, enterochromaffin (EC) cell number, expression of TPH, and faecal/colonic 5-HT contents were measured. RESULTS: Loose stool and small intestine transit rates, EC cell numbers, colonic TPH expression and faecal/colonic 5-HT content of IBS-D rats were significantly increased relative to controls (p<0.05) and all these parameters were improved by EA at ST25, BL25, or ST25 and BL25 in combination (all p<0.05 vs untreated IBS-D rats). CONCLUSIONS: EA at ST25 and/or BL25 had a positive effect on objective markers of diarrhoea in a IBS-D rat model and induced changes in EC cell number, colonic TPH and 5-HT contents. The effects of EA stimulation at ST25/BL25 on IBS-D rats may be mediated by excitation of sympathetic nerves.


Assuntos
Pontos de Acupuntura , Diarreia/terapia , Eletroacupuntura , Síndrome do Intestino Irritável/terapia , Animais , Diarreia/metabolismo , Modelos Animais de Doenças , Humanos , Síndrome do Intestino Irritável/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Serotonina/metabolismo
9.
Acupunct Med ; 34(3): 194-200, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26561562

RESUMO

BACKGROUND: Electroacupuncture (EA) is used clinically for the treatment of constipation. Serotonin (5-hydroxytryptamine, 5-HT) plays an important role in colonic motility; however it is unknown whether alterations in colonic 5-HT are associated with EA. In this study, the effect and mechanism of EA at acupuncture points LI11 and ST37 were examined using a cold saline-induced rat model of constipation. METHODS: A rat constipation model was induced by cold saline gavage in 24 Sprague-Dawley rats. A further six rats were included as a Control group. The constipated rats were divided into four groups (n=6 each): a Constipation group that remained untreated; a Constipation+LI11 group that received EA at LI11; a Constipation+ST37 groups that received EA at ST37; and a Constipation+LI11+ST37 group that received EA at both LI11 and ST37. After EA treatment, faecal water content, defaecation frequency, and gastrointestinal (GI) transit were measured, as well as the expression of tryptophan hydroxylase (TPH) in colonic tissues (by Western blot analysis) and 5-HT in both faeces and colonic tissues (by ELISA). RESULTS: All three EA-treated groups demonstrated significant improvements in faecal water content, defaecation frequency and GI transit (p<0.05). In addition, TPH and 5-HT expression were both increased by EA at LI11 and/or ST37 (p<0.05). There were no significant differences between the three EA groups for any outcomes. CONCLUSIONS: EA at LI11 and/or ST37 had a positive effect on objective markers of constipation in a rat model. In addition, EA increased 5-HT and TPH in the colonic tissues.


Assuntos
Pontos de Acupuntura , Constipação Intestinal/terapia , Eletroacupuntura , Animais , Constipação Intestinal/metabolismo , Defecação , Fezes , Trânsito Gastrointestinal , Masculino , Ratos , Ratos Sprague-Dawley , Serotonina/metabolismo , Triptofano Hidroxilase/metabolismo
11.
J Allergy Clin Immunol ; 136(2): 454-61.e9, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25748343

RESUMO

BACKGROUND: The mechanisms underlying glucocorticoid responsiveness are largely unknown. Although redox regulation of the glucocorticoid receptor (GR) has been reported, it has not been studied in asthmatic patients. OBJECTIVE: We characterized systemic cysteine oxidation and its association with inflammatory and clinical features in healthy children and children with difficult-to-treat asthma. We hypothesized that cysteine oxidation would be associated with increased markers of oxidative stress and inflammation, increased features of asthma severity, decreased clinically defined glucocorticoid responsiveness, and impaired GR function. METHODS: PBMCs were collected from healthy children (n = 16) and children with asthma (n = 118) aged 6 to 17 years. Children with difficult-to-treat asthma underwent glucocorticoid responsiveness testing with intramuscular triamcinolone. Cysteine, cystine, and inflammatory chemokines and reactive oxygen species generation were quantified, and expression and activity of the GR were assessed. RESULTS: Cysteine oxidation was present in children with difficult-to-treat asthma and accompanied by increased reactive oxygen species generation and increased CCL3 and CXCL1 mRNA expression. Children with the greatest extent of cysteine oxidation had more features of asthma severity, including poorer symptom control, greater medication use, and less glucocorticoid responsiveness despite inhaled glucocorticoid therapy. Cysteine oxidation also modified the GR protein by decreasing available sulfhydryl groups and decreasing nuclear GR expression and activity. CONCLUSIONS: A highly oxidized cysteine redox state promotes a posttranslational modification of the GR that might inhibit its function. Given that cysteine oxidation is prevalent in children with difficult-to-treat asthma, the cysteine redox state might represent a potential therapeutic target for restoration of glucocorticoid responsiveness in this population.


Assuntos
Asma/tratamento farmacológico , Glucocorticoides/uso terapêutico , Leucócitos Mononucleares/imunologia , Processamento de Proteína Pós-Traducional , Receptores de Glucocorticoides/imunologia , Triancinolona/uso terapêutico , Administração por Inalação , Adolescente , Asma/genética , Asma/imunologia , Asma/patologia , Quimiocina CCL3/genética , Quimiocina CCL3/imunologia , Quimiocina CXCL1/genética , Quimiocina CXCL1/imunologia , Criança , Cisteína/química , Cisteína/imunologia , Cistina/química , Cistina/imunologia , Monitoramento de Medicamentos , Feminino , Expressão Gênica , Humanos , Injeções Intramusculares , Leucócitos Mononucleares/química , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/patologia , Masculino , Oxirredução , Estresse Oxidativo , Cultura Primária de Células , RNA Mensageiro/genética , RNA Mensageiro/imunologia , Receptores de Glucocorticoides/química , Receptores de Glucocorticoides/genética
12.
Behav Brain Res ; 280: 160-71, 2015 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-25449842

RESUMO

Schizophrenia is thought to be caused, at least in part, by dysfunction in striatal dopamine neurotransmission. Both clinical studies and animal research have implicated the dopamine neuromodulator neurotensin (NT) in the pathophysiology of schizophrenia. Utilizing male mice lacking the NT gene (NT(-/-)), these studies examined the consequences of NT deficiency on dopaminergic tone and function, investigating (1) dopamine concentrations and dopamine receptor and transporter expression and binding in dopaminergic terminal regions, and (2) the behavioral effects of selective dopamine receptor agonists on locomotion and sensorimotor gating in adult NT(-/-) mice compared to wildtype (NT(+/+)) mice. NT(-/-) mice did not differ from NT(+/+) mice in concentrations of dopamine or its metabolite DOPAC in any brain region examined. However, NT(-/-) mice showed significantly increased D1 receptor, D2 receptor, and dopamine transporter (DAT) mRNA in the caudate putamen compared to NT(+/+) controls. NT(-/-) mice also showed elevated D2 receptor binding densities in both the caudate putamen and nucleus accumbens shell compared to NT(+/+) mice. In addition, some of the behavioral effects of the D1-type receptor agonist SKF-82958 and the D2-type receptor agonist quinpirole on locomotion, startle amplitude, and prepulse inhibition were dose-dependently altered in NT(-/-) mice, showing altered D1-type and D2-type receptor sensitivity to stimulation by agonists in the absence of NT. The results indicate that NT deficiency alters striatal dopamine receptor expression, binding, and function. This suggests a critical role for the NT system in the maintenance of striatal DA system homeostasis and implicates NT deficiency in the etiology of dopamine-associated disorders such as schizophrenia.


Assuntos
Corpo Estriado/fisiologia , Proteínas da Membrana Plasmática de Transporte de Dopamina/metabolismo , Plasticidade Neuronal/fisiologia , Neurotensina/deficiência , Receptores de Dopamina D1/metabolismo , Receptores de Dopamina D2/metabolismo , Ácido 3,4-Di-Hidroxifenilacético/metabolismo , Animais , Benzazepinas/farmacologia , Corpo Estriado/efeitos dos fármacos , Dopamina/metabolismo , Agonistas de Dopamina/farmacologia , Masculino , Camundongos Knockout , Atividade Motora/efeitos dos fármacos , Atividade Motora/fisiologia , Plasticidade Neuronal/efeitos dos fármacos , Neurotensina/genética , Quimpirol/farmacologia , RNA Mensageiro/metabolismo , Reflexo de Sobressalto/efeitos dos fármacos , Reflexo de Sobressalto/fisiologia , Filtro Sensorial/efeitos dos fármacos , Filtro Sensorial/fisiologia
13.
Zhongguo Zhen Jiu ; 34(5): 449-53, 2014 May.
Artigo em Chinês | MEDLINE | ID: mdl-25022114

RESUMO

OBJECTIVE: To verify the clinical efficacy of acupuncture combined with auricle cutting method for treatment of blood stasis-type psoriasis. METHODS: Fifty-six cases of blood stasis-type psoriasis were randomly divided into a combined therapy group, a auricle cutting group, an acupuncture group and a control group, 14 cases in each one. Based on regular treatment of TCM decoction in four groups, the combined therapy group was treated with acupuncture and auricle cutting method, and the auricle cutting group was treated with sham-acupuncture and auricle cutting, and the acupuncture group was treated with acupuncture and sham auricle cutting, and the control group was treated with sham-acupuncture and sham auricle cutting. The acupuncture was applied at Dazhui (GV 14), Feishu (BL 13), Ganshu (BL 18) and Geshu (BL 17), etc., and manipulated with routine technique; in the sham acupuncture, the needle was inserted into dermis layer so that the needles could be swung without being dropped out. In the auricle cutting, erbeixin (P1) of unilateral auricle was selected and cut by Chan needle to perform bloodletting; in the sham auricle cutting, the neighborhood approximately 0.5 cm next to erbeixin (P) of auricle was selected as cutting area. The treatment was given once a day, seven days as a treatment session for totally two sessions. Psoriasis area and severity index (PASI) before and after treatment was observed and efficacy of each group was compared. RESULTS: The effective rate was 57.1% (8/14) in the combined therapy group, which was superior to 14.3% (2/14) in the auricle cutting group, 7.1% (1/14) in the acupuncture group and 0.0% (0/14) in the control group (all P < 0.05). The scores of PASI were all decreased in each group after the treatment (all P < 0.05), which was the most significant in the combined therapy group (all P < 0.05). After factorial analysis, the main effect was P < 0.05 in the auricle cutting, P < 0.05 in the acupuncture and P < 0.05 in interaction effect of combined therapy. CONCLUSION: The scores of PASI of blood stasis-type psoriasis could be effectively reduced by acupuncture, auricle cutting method and TCM decoction, among which the interaction effect of auricle cutting and acupuncture combined with TCM decoction is the most significant.


Assuntos
Terapia por Acupuntura , Sangria , Orelha/irrigação sanguínea , Psoríase/terapia , Adolescente , Adulto , Idoso , Terapia Combinada , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Psoríase/sangue , Resultado do Tratamento , Adulto Jovem
14.
Appl Microbiol Biotechnol ; 98(18): 7837-44, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24728756

RESUMO

Pramlintide is an artificially designed protein which has the same function as amylin in human body. This protein is extremely difficult to synthesize through prokaryotic expression method because of its two essential active sites, intrachain disulfide bond and C-terminal amide group. Since α-amidating monooxygenase is widely distributed in human and animal, it is possible to use pramlintide precursor with an additional C-terminal glycine (PAG), which is the potential substrate of α-amidating monooxygenase, for in vivo applications. The recombinant PAG was expressed in Escherichia coli using the small ubiquitin-related modifier (SUMO) as the molecular chaperone, and the optimal fusion expression level reached to 36.3% of the total supernatant protein. Under optimal conditions in a 10-L fermentor, the recombinant PAG was obtained with a purity of greater than 95%, and the average expression level was reached to 20 mg/L. The authenticity and the intrachain disulfide bridge of PAG were confirmed by Western blotting and matrix-assisted laser desorption/ionization coupled to time-of-flight mass spectrometry (MALDI-TOF MS) as well as N-terminal sequencing of protein. Based on an L6 myoblast cell model in vitro and an animal model of gastric emptying in vivo, the results of activity revealed that PAG showed a lower biological activity in vitro but has almost the same activity as the chemically synthesized pramlintide in vivo.


Assuntos
Escherichia coli/metabolismo , Polipeptídeo Amiloide das Ilhotas Pancreáticas/metabolismo , Polipeptídeo Amiloide das Ilhotas Pancreáticas/farmacologia , Proteínas Recombinantes de Fusão/metabolismo , Proteínas Recombinantes de Fusão/farmacologia , Animais , Linhagem Celular , Esvaziamento Gástrico/efeitos dos fármacos , Polipeptídeo Amiloide das Ilhotas Pancreáticas/genética , Masculino , Ratos , Ratos Sprague-Dawley , Proteínas Recombinantes de Fusão/genética
15.
ScientificWorldJournal ; 2014: 806371, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24723829

RESUMO

To investigate the local micro-/nanoscale region in a large scale sample, an image reconstruction method for nanometer computed tomography (nano-CT) was proposed in this paper. In the algorithm, wavelets were used to localize the filtered-backprojection (FBP) algorithm because of its space-frequency localization property. After the implementation of the algorithm, two simulation local reconstruction experiments were performed to confirm its effectiveness. Three evaluation criteria were used in the experiments to judge the quality of the reconstructed images. The experimental results showed that the algorithm proposed in this paper performed best because (1) the quality of its results had improved 20%-30% compared to the results of FBP and 10%-30% compared to the results of another wavelet algorithm; (2) the new algorithm was stable under different circumstances. Besides, an actual reconstruction experiment was performed using real projection data that had been collected in a CT experiment. Two-dimensional (2D) and three-dimensional (3D) images of the sample were reconstructed. The microstructure of the sample could be clearly observed in the reconstructed images. Since much attention has been directed towards the nano-CT technique to investigate the microstructure of materials, this new wavelet-based local tomography algorithm could be considered as a meaningful effort.


Assuntos
Algoritmos , Diagnóstico por Imagem/métodos , Processamento de Imagem Assistida por Computador/métodos , Imagem Tridimensional , Tomografia Computadorizada por Raios X
16.
Oxid Med Cell Longev ; 2013: 543760, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24454985

RESUMO

OBJECTS: The aim of this study is to evaluate protein oxidation, DNA damage, and lipid peroxidation in patients with gastric cancer and to investigate the relationship between oxidative stress and gastric cancer. METHODS: We investigated changes in serum protein carbonyl (PC), advanced oxidation protein products (AOPP), and 3-nitrotyrosine (3-NT) levels, as indicators of protein oxidation, serum 8-hydroxydeoxyguanosine (8-OHdG), as a biomarker of DNA damage, and malondialdehyde (MDA), conjugated diene (CD), 4-hydroxynonenal (4-HNE), and 8-ISO-prostaglandin F2α (8-PGF) in serum, as lipid peroxidation markers in gastric cancer (GC) patients and healthy control. RESULTS: Compared with control, a statistically significant higher values of 8-OHdG, PC, AOPP, and 3-NT were observed in the GC patients (P < 0.05). The products of lipid peroxidation, MDA, CD, 4-HNE, and 8-PGF, were significantly lower in the GC patients compared to those of control (P < 0.05). In addition, the products of oxidative stress were similar between the Helicobacter pylori positive and the negative subgroups of GC patients. CONCLUSIONS: GC patients were characterized by increased protein oxidation and DNA damage, and decreased lipid peroxidation. Assessment of oxidative stress and augmentation of the antioxidant defense system may be important for the treatment and prevention of gastric carcinogenesis.


Assuntos
Dano ao DNA , Peroxidação de Lipídeos , Proteínas de Neoplasias/metabolismo , Neoplasias Gástricas/metabolismo , Estudos de Casos e Controles , Demografia , Feminino , Infecções por Helicobacter/sangue , Infecções por Helicobacter/metabolismo , Infecções por Helicobacter/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Oxirredução , Estresse Oxidativo , Neoplasias Gástricas/sangue
17.
Zhongguo Zhen Jiu ; 31(9): 773-7, 2011 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-21972616

RESUMO

OBJECTIVE: To observe the clinical efficacy of electroacupuncture (EA) on mild cognitive impairment (MCI) for patients of Uygur and Han nationality and explore the national diversity among the patients with MCI. METHODS: Twenty-five cases were divided into Han nationality group (15 cases) and Uygur nationality group (10 cases) according to patient's nationality. In either group, EA was applied to Baihui (GV 20), Fengchi (GB 20), Xuanzhong (GB 39), Fuliu (KI 7), Sanyinjiao (SP 6) and Taixi (KI 3), once per day, 15 treatments made one session and there were 5 days at the interval among the sessions. Totally, 3 sessions of treatment were required. The proton magnetic resonance spectroscopy (1H-MRS) was used to observe the changes in the ratio of N-acetylaspartate and creatine (NAA/Cr) on the left hippocampus for the patients in two groups before and after treatment as well as the changes in the results of the Mini-Mental State Examination (MMSE) and the Montreal Cognitive Assessment (MoCA) separately. RESULTS: NAA/Cr in Uygur nationality group was higher than that in Han nationality group before treatment (1.659 +/- 0.418 vs 1.137 +/- 0.190, P < 0.05). After treatment, MMSE and MoCA scores all increased apparently as compared with those before treatment in two groups (P < 0.05, P < 0.01), and NAA/Cr on the left hippocampus in either group was up-regulated as compared with that before treatment (both P < 0.01). CONCLUSION: EA can improve the overall cognitive function for the patients with MCI. There is the national diversity in the partial brain metabolite level between Uygur patients and Han patients with MCI.


Assuntos
Ácido Aspártico/análogos & derivados , Transtornos Cognitivos/terapia , Creatina/análise , Hipocampo/química , Idoso , Idoso de 80 Anos ou mais , Ácido Aspártico/análise , China/etnologia , Transtornos Cognitivos/metabolismo , Eletroacupuntura , Feminino , Humanos , Espectroscopia de Ressonância Magnética , Masculino , Pessoa de Meia-Idade
18.
Cancer Invest ; 28(6): 672-8, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20394500

RESUMO

ZBTB7 was recently recognized as a proto-oncogene. We studied its prognostic value and relationship to clinicopathological variables in 125 breast cancer patients. ZBTB7 expression was significantly higher in breast cancer tissues than in normal breast tissues, but its gene amplification copies were relatively low. ZBTB7 expression levels were significantly correlated with histological grade (p = .023) and marginally inversely correlated with the presence of estrogen receptors (p = .053); its overexpression significantly predicted shorter recurrence-free survival (p = .033). Our results showed that ZBTB7 might be implicated in breast cancer development and may serve as a promising prognostic marker.


Assuntos
Biomarcadores Tumorais/análise , Neoplasias da Mama/química , Proteínas de Ligação a DNA/análise , Fatores de Transcrição/análise , Biomarcadores Tumorais/genética , Western Blotting , Neoplasias da Mama/genética , Neoplasias da Mama/mortalidade , Neoplasias da Mama/patologia , Neoplasias da Mama/terapia , Distribuição de Qui-Quadrado , Proteínas de Ligação a DNA/genética , Intervalo Livre de Doença , Feminino , Amplificação de Genes , Regulação Neoplásica da Expressão Gênica , Humanos , Imuno-Histoquímica , Estimativa de Kaplan-Meier , Estadiamento de Neoplasias , Reação em Cadeia da Polimerase , Receptores Estrogênicos/análise , Recidiva , Medição de Risco , Fatores de Tempo , Fatores de Transcrição/genética , Resultado do Tratamento , Regulação para Cima
19.
Gynecol Oncol ; 110(2): 230-6, 2008 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-18538832

RESUMO

OBJECTIVE: Nlp (Ninein-like protein), a novel centrosome protein involved in microtubule nucleation, has been studied extensively in our laboratory, and its overexpression has been found in some human tumors. To understand the role of Nlp in human ovarian cancer development, we studied the correlation of Nlp expression with clinicopathological parameters and survival in epithelial ovarian cancer, and the impact of Nlp overexpression on ovarian cancer cells. METHODS: Nlp expression in normal, borderline, benign and malignant epithelial ovarian tissues was examined by immunohistochemistry. The correlation between Nlp expression and tumor grade, FIGO stage and histological type was also evaluated. Survival was calculated using Kaplan-Meier estimates. Cell proliferation and apoptosis were assayed after stable transfection of pEGFP-C3-Nlp or empty vector in human ovarian cancer cell line SKOV3. RESULTS: Nlp was positive in 1 of 10 (10%) normal ovarian tissues, 5 of 34 (14.7%) benign tumors, 9 of 26 (34.6%) borderline tumors and 73 of 131 (56.0%) ovarian tumors. Nlp immunoreactivity intensity significantly correlated with tumor grade, but not with FIGO stage or histological type. Kaplan-Meier curves showed that Nlp overexpression was marginally associated with decreased overall survival. Overexpression of Nlp enhanced proliferation and inhibited apoptosis induced by paclitaxel in the SKOV3 cell line. CONCLUSIONS: Overexpression of Nlp in ovarian tumors raises the possibility that Nlp may play a role in ovarian carcinogenesis.


Assuntos
Adenocarcinoma/metabolismo , Transformação Celular Neoplásica/metabolismo , Proteínas Associadas aos Microtúbulos/biossíntese , Proteínas Nucleares/biossíntese , Neoplasias Ovarianas/metabolismo , Adenocarcinoma/genética , Adenocarcinoma/patologia , Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Apoptose/fisiologia , Processos de Crescimento Celular/fisiologia , Linhagem Celular Tumoral , Transformação Celular Neoplásica/genética , Feminino , Humanos , Imuno-Histoquímica , Proteínas Associadas aos Microtúbulos/genética , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Proteínas Nucleares/genética , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/patologia , Paclitaxel/farmacologia , Taxa de Sobrevida , Transfecção
20.
Zhongguo Fei Ai Za Zhi ; 11(3): 368-72, 2008 Jun 20.
Artigo em Chinês | MEDLINE | ID: mdl-20731936

RESUMO

BACKGROUND: X-linked inhibitor of apoptosis protein (XIAP) is a newly discovered inhibitor of apoptosis protein which prevents apoptosis by inhibiting the activation of caspase. After down-regulating XIAP gene expression in A549 cells, a non-small cell lung cancer (NSCLC) cell lines, we investigated the role of XIAP specific siRNA in apoptosis and chemotherapy sensitivity. METHODS: The mRNA levels of XIAP gene in A549 cells were assessed using a semi-quantitative reverse transcriptase-PCR (RT-PCR). The expression vector of XIAP small interfering RNA (XIAP siRNAwas constructed and transfected into A549 cells. The transfection was proved effective by the fluorescence microscope. Cell proliferation and cell killing rate after chemotherapeutics treatment were investigated by MTT assay. The rate of apoptosis was detected by flow cytometry assay. RESULTS: XIAP siRNA construction was proved successful by enzyme digestion and DNA sequencing. The transfection efficiency in A549 cells from positive transfection group and negative transfection group had no differences. Compared to those in cell from control group, the level of XIAP mRNA expression was significantly decreased, the inhibition activity of Cisplatin was significantly higher in cells from positive transfection group. Proliferation of cells from positive transfection group was significantly inhibited after 24, 48, 72, 96 hours . The rates cell killing and apoptosis in cells from positive transfection group caused by Cisplatin were significantly higher compared to those cells from control group. CONCLUSIONS: The increased expression of XIAP in NSCLC can inhibit the apoptosis of NSCLC cells and result in NSCLC chemotherapy drug resistance. XIAP siRNA could inhibit the NSCLC cell growth specifically, down-regulation of XIAP gene expression promote apoptosis and increase the chemotherapy sensitivity of NSCLC. XIAP siRNA sequence might become a therapeutic target of NSCLC.

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