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1.
Microb Pathog ; 142: 104077, 2020 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-32084579

RESUMO

Alpha-linolenic acid (ALA), an important component of polyunsaturated fatty acids (PUFAs), possesses potent anti-inflammatory properties. To date, the effects of ALA on acute lung injury (ALI) remains unknown. This study was designed to investigate the potential protective effects of ALA on LPS-induced ALI and the underpinning mechanisms. An animal model of ALI was established via intratracheally injection of lipopolysaccharide (LPS, 1 mg/kg). We found that lung wet/dry weight ratio and protein concentration in Bronchoalveolar lavage fluid (BALF) were dramatically decreased by ALA pretreatment. Treatment with ALA significantly alleviated the infiltration of total cells and neutrophils, while increased the number of the macrophages. ALA significantly inhibited the secretion of proinflammatory cytokines including tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1ß (IL-1ß) and increased anti-inflammatory cytokine. Moreover, we found that the levels of myeloperoxidase (MPO) and malondialdehyde (MDA) were highly increased in LPS-induced ALI, while the activities of glutathione (GSH) and superoxide dismutase (SOD) were decreased, which were reversed by ALA. ALA attenuated LPS-induced histopathological changes and apoptosis. Furthermore, ALA significantly inhibited the phosphorylation of IκBα and NF-κB (p65) activation in ALI. ALA showed anti-inflammatory effects in mice with LPS-induced ALI. NF-κB pathway may be involved in ALA mediated protective effects.

2.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 51(1): 67-73, 2020 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-31950792

RESUMO

Objective: To develop and verify a flow cytometric measurement of reticulocytes (RETs) micronucleus in rat bone marrow. Methods: In our flow cytometric protocol, reticulocytes, leukocytes and DNA were labeled by anti-CD71-fluorescein isothiocyanate (FITC), anti-CD45-phycoerythrin (PE) and DRAQ5, respectively. Sprague-Dawley (SD) rats were assigned to four treatment groups randomly, and were exposed to ethyl methanesulfonate (EMS), cyclophosphamide (CP), ethyl nitrosourea (ENU) and colchicine (COL) respectively. Each treatment group was divided into four subgroups (5 rats per subgroup) according to different exposure dosage. A exposure dose of 0 was used as vehicle control for each group. Rats were administered with testing mutagens by gavage twice with a 24 h interval. Bone marrow from both femurs were collected 24 h after the last administration. The frequency of micronucleated reticulocytes (MN-RETs) and the percentage of reticulocytes (RETs%) were determined by flow cytometric measurement established in this study. And the manual counting method with microscope (by Giemsa staining) was conducted at the same time. Results: A method for detection of reticulocyte micronucleus in bone marrow based on flow cytometry was successfully established. The MN-RETs in rat bone marrow of 20 SD rats treated by vehicle (i.e., background value of MN-RETs) was 0.83‰±0.12‰ by this method. The background value of MN-RETs in manual enumeration method was 1.43‰±0.44‰. It was obvious that the flow cytometric method had lower background value and more stable results. The trend, in which MN-RETs ascended and RETs% descended with increasing dose, can be detected by both methods in rats that exposed to EMS, CP, ENU and COL. Both methods were good to detect the correlation of induced-MN-RETs with four testing mutagens (the correlation coefficients were ranged from 0.834 3 to 0.913 7). Conclusion: With its sensitivity, rapidity, easy operation and low background value, the three-color flow cytometric enumerative protocol established in our laboratory can be used as a good substitute for manual micronucleus counting method and used in genotoxicity assessment of chemical substances.


Assuntos
Medula Óssea , Citometria de Fluxo , Reticulócitos , Animais , Testes para Micronúcleos , Ratos , Ratos Sprague-Dawley , Reticulócitos/citologia
3.
Wei Sheng Yan Jiu ; 48(6): 976-1000, 2019 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-31875825

RESUMO

OBJECTIVE: This study was designed to determine the genotoxicity of 2-methylfuran based on a multi-endpoint genotoxicity test system. METHODS: The SPF-grade male SD rats(n = 30) were randomized to six treatment groups, i. e. 4 treatment groups(25, 50, 100 and 150 mg/kg), a control group(vegetable oil) and a positive groups(N-ethyl-N-nitrosourea, 80 mg/kg). All treatments were administrated by gavage for continuous 3 days. Tail vein blood for comet assay was collected at 3 h after the final administration. Pig-a gene mutation assays were performed on days 0(one day before gavage), 14 and 28. Micronucleus tests in peripheral blood using flow cytometry were performed on days 0 and 4. RESULTS: A statistically significant increase in tail intensity was observed at 150 mg/kg for peripheral blood in comet assay. There was no significant difference among the groups in mutant cell frequency of erythrocytes and reticulocytes at 2 timepoints in Pig-a gene mutation assay, and no significant difference among the groups in the frequency of micronucleus in micronucleus test. CONCLUSION: The result of genotoxicity tests suggested that 2-methylfuran was probably not mutagenic in vivo after acute exposure.


Assuntos
Eritrócitos , Animais , Ensaio Cometa , Furanos , Masculino , Testes para Micronúcleos , Testes de Mutagenicidade , Ratos , Ratos Sprague-Dawley
4.
Appl Microbiol Biotechnol ; 103(21-22): 8825-8838, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31637492

RESUMO

Microbial-induced CaCO3 precipitation has been widely applied in bacterial-based self-healing concrete. However, the limited biogenetic CaCO3 production by bacteria after they were introduced into the incompatible concrete matrix is a major challenge of this technology. In the present study, the potential of combining two metabolic pathways, urea hydrolysis and nitrate reduction, simultaneously in one bacteria strain for improving the bacterial CaCO3 yield has been investigated. One bacterial strain, Ralstonia eutropha H16, which has the highest Ca2+ tolerance and is capable of performing both urea hydrolysis and nitrate reduction in combined media was selected among three bacterial candidates based on the enzymatic examinations. Results showed that H16 does not need oxygen for urea hydrolysis and urease activity was determined primarily by cell concentration. However, the additional urea in the combined medium slowed down the nitrate reduction rate to 7 days until full NO3- decomposition. Moreover, the nitrate reduction of H16 was significantly restricted by an increased Ca2+ ion concentration in the media. Nevertheless, the overall CaCO3 precipitation yield can be improved by 20 to 30% after optimization through the combination of two metabolic pathways. The highest total CaCO3 precipitation yield achieved in an orthogonal experiment was 14 g/L. It can be concluded that Ralstonia eutropha H16 is a suitable bacterium for simultaneous activation of urea hydrolysis and nitrate reduction for improving the CaCO3 precipitation and it can be studied later, on activation of multiple metabolic pathways in bacteria-based self-healing concrete.

5.
Medicine (Baltimore) ; 98(39): e17209, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31574830

RESUMO

BACKGROUND: This study is to summarize the status of knowledge, attitudes, implementation, facilitators, and barriers of evidence-based practice (EBP) in community nurses (CNs). EBP has been widely adopted but the knowledge, attitudes, and implementation of EBP among CNs, and the facilitators and barriers they perceived have not been clearly confirmed. METHODS: A literature search was conducted using combined keywords in 3 English databases and 3 Chinese databases of peer-reviewed publications covering the dates of publication from 1996 to July, 2018. Twenty articles were included. The information of the knowledge, attitudes, implementation, and the perceived facilitators and barriers of EBP in CNs was extracted and summarized. RESULTS: CNs had positive attitudes toward EBP, but insufficient knowledge and unprepared implementation. The most cited facilitators were academic training, management functions, and younger age. Inadequate time and resources were recognized as main barriers hindering the transforming from knowledge and attitudes to implementation. Developed interventions mainly focused on knowledge facilitation rather than the elimination of objective barriers. CONCLUSIONS: Findings demonstrate a compelling need for improvement in knowledge and implementation of EBP in CNs, compared with the better attitudes. Except education, knowledge translating into implementation needs more coordination with authorities to magnify the facilitators and overcome the barriers. Further studies need to concentrate on deficient knowledge and implementation of EBP among CNs. Policy makers can use the facilitators and barriers found by this review to modify nursing education, current scientific resources supplement, practice supports for care improving.


Assuntos
Atitude do Pessoal de Saúde , Prática Clínica Baseada em Evidências , Conhecimentos, Atitudes e Prática em Saúde , Implementação de Plano de Saúde , Enfermeiras de Saúde Comunitária/psicologia , Adulto , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
6.
J Mol Cell Biol ; 2019 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-30925593

RESUMO

There are intense interests in discovering proregenerative medicine leads that can promote cardiac differentiation and regeneration, as well as repair damaged heart tissues. We have combined zebrafish embryo-based screens with cardiomyogenesis assays to discover selective small molecules that modulate heart development and regeneration with minimal adverse effects. Two related compounds with novel structures, named as Cardiomogen1 and 2 (CDMG1 and CDMG2), were identified for their capacity to promote myocardial hyperplasia through expansion of the cardiac progenitor cell population. We find that Cardiomogen acts as a Wnt inhibitor by targeting ß-catenin and reducing Tcf/Lef-mediated transcription in cultured cells. CDMG treatment of amputated zebrafish hearts reduces nuclear ß-catenin in injured heart tissue, increases cardiomyocyte (CM) proliferation, and expedites wound healing, thus accelerating cardiac muscle regeneration. Importantly, Cardiomogen can alleviate the functional deterioration of mammalian hearts after myocardial infarction. Injured hearts exposed to CDMG1 display increased newly formed CMs and reduced fibrotic scar tissue, which are in part attributable to the ß-catenin reduction. Our findings indicate Cardiomogen as a Wnt inhibitor in enhancing injury-induced CM proliferation and heart regeneration, highlighting the values of embryo-based small molecule screens in discovery of effective and safe medicine leads.

7.
Mol Ther Nucleic Acids ; 16: 1-14, 2019 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-30825668

RESUMO

miR-181a is downregulated in leukemia and affects its progression, drug resistance, and prognosis. However, the exact mechanism of its targets in leukemia, particularly in chronic myelogenous leukemia (CML), has not previously been established. Here, we use a multi-omics approach to demonstrate that protein tyrosine phosphatase, receptor type, f polypeptide, leukocyte common antigen (LAR) interacting protein (liprin), alpha 1 (PPFIA1) is a direct target for miR-181a in CML. Phospho-array assay shows that multiple phosphorylated proteins, particularly KIT signaling molecules, were downregulated in PPFIA1 inhibition. Additionally, PPFIA1 bound PARP1, a common molecule downstream of both PPFIA1 and BCR/ABL, to upregulate KIT protein through activation of nuclear factor kappa B (NF-κB)-P65 expression. Targeted inhibition of PPFIA1 and PARP1 downregulated c-KIT level, inhibited CML cell growth, and prolonged mouse survival. Overall, we report a critical regulatory miR-181a/PPFIA1/PARP1/NF-κB-P65/KIT axis in CML, and our preclinical study supports that targeted PPFIA1 and PARP1 may serve as a potential CML therapy.

8.
Wei Sheng Yan Jiu ; 47(4): 525-553, 2018 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-30081975

RESUMO

OBJECTIVE: To make recommendations on the design and analysis of Piga gene mutation assay on the basis of the historical control data. METHODS: All negative historical control data( total: 128) were collected to create the historical control data base, and descriptive statistics was used to determine the distribution of the data. The power of the number of animals per group, number of measured cells per animal and number of groups were discussed through simulation test based on historical data base. RESULTS: The group mean of mutant red blood cells( RBCs~(CD59-)) and mutant reticulocytes ( RETs~(CD59-)) were 26. 63 × 10~(-6) and 35. 85 × 10~(-6), respectively, and the standard deviation were 27. 71 × 10~(-6) and 31. 06 × 10~(-6), respectively. By mapping the frequencies of data, the variables had skewed distributions and translated to normal distributions after logarithmic transformation. The confidence level for population means were 100% and 92% for RBCs( 1 × 10~6 cells per animal) and RETs( 0. 3 × 10~6 cells per animal), and it increased to 100% when 1 × 10~6 cells was scored for RETs. Group sizes of 5 had low statistical power while the minimal detectable difference was 2 times. The power had increased to > 80% when 4- or 5- fold of minimal detectable difference was employed. CONCLUSION: In brief, the recommendations include:① A log( 10) transformation of mutant frequencies often has been found satisfactory for data when parametric method such as an analysis of variance are used. ② Young adult animals( approximately 6 weeks of age rats) are recommended in this assay. ③ The recommended number of RETs and RBCs are > 1 × 10~6 and > 5 × 10~7, respectively. ④ Based on power calculations, 3 treatment groups and 1 negative control group are appropriate when fourfold increase was employed. ⑤ Group sizes of 6 or 7 are recommended but 5 analyzable animals per group may be acceptable when 4 test groups were used.


Assuntos
Eritrócitos/efeitos dos fármacos , Proteínas de Membrana/genética , Mutagênicos/toxicidade , Mutação/genética , Reticulócitos/efeitos dos fármacos , Animais , Eritrócitos/metabolismo , Testes de Mutagenicidade/métodos , Ratos , Reticulócitos/metabolismo
9.
Front Microbiol ; 9: 390, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29593670

RESUMO

Porcine circovirus-like virus P1 is an important pathogen of the current pig industry, the infection mechanism is not entirely clear. Wnt signaling pathway plays an important role in the growth of young animals and infection of some viruses. This study was designed to demonstrate the effects of P1 infection on the Wnt signaling pathway. In vivo experiments, we demonstrated the down-regulatory effects of P1 infection in piglets and mice on the downstream components expression levels of Wnt signaling pathway, and the effects of Wnt signaling pathway activation on the pathogenesis of P1. In vitro studies, we found P1 infection down-regulated protein level of ß-catenin and mRNA level of mmp2, prevented the ß-catenin from entering into nucleus, abolished the TCF/LEF promoter activity, proved that P1 could inhibit the activation of Wnt signaling pathway in vitro. Finally, we found that VP1 of P1 virus also had the inhibitory effects on Wnt signaling pathway in vitro, elucidated the mechanism of P1's inhibitory effects on the Wnt signaling pathway and offered the possibility that the suppression of Wnt signaling pathway was involved in the post-weaning multisystemic wasting syndrome (PMWS), laying a foundation for elucidating the pathogenesis of P1.

10.
Int J Nurs Stud ; 77: 171-178, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29100199

RESUMO

BACKGROUND: The hypertension prevalence rate is increasing but the control rate is unsatisfactory. Nurse-led healthcare may be an effective way to improve outcomes for hypertensive patients but more evidence is required especially at the community level. OBJECTIVE: This study aims to establish a nurse-led hypertension management model and to test its effectiveness at the community level. DESIGN: A single-blind, randomized controlled trial was performed in an urban community healthcare center in China. Hypertensive patients with uncontrolled blood pressure (systolic blood pressure ≥140mmHg and/or diastolic blood pressure ≥90mmHg) were randomly allocated into two groups: the study group (n=67) and the control group (n=67). The nurse-led hypertension management model included four components (delivery system design, decision support, clinical information system and self-management support). Patients in the control group received usual care. Patients in the study group received a 12-week period of hypertension management. The patient outcomes, which involved blood pressure, self-care behaviors, self-efficacy, quality of life and satisfaction, were assessed at three time points: the baseline, immediately after the intervention and 4 weeks after the intervention. RESULTS: After the intervention, the blood pressure of patients in the study group decreased significantly compared to those in the control group, and the mean reduction of systolic/diastolic blood pressure in the study and control groups was 14.37/7.43mmHg and 5.10/2.69mmHg, respectively (p<0.01). In addition, patients in the study group had significantly greater improvement in self-care behaviors than those in the control group (p<0.01). The study group had a higher level of satisfaction with hypertensive care than the control group (p<0.01). No statistically significant difference in self-efficacy and quality of life was detected between the two groups after the intervention. CONCLUSIONS: The nurse-led hypertension management model is feasible and effective in improving the outcomes of patients with uncontrolled blood pressure at the community level.


Assuntos
Hipertensão/terapia , Relações Enfermeiro-Paciente , Idoso , Pressão Sanguínea , China , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Satisfação do Paciente , Qualidade de Vida , Autocuidado , Autoeficácia , Método Simples-Cego
11.
J Int Med Res ; 45(1): 22-37, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28222625

RESUMO

Objective Routine fasting (12 h) is always applied before laparoscopic cholecystectomy, but prolonged preoperative fasting causes thirst, hunger, and irritability as well as dehydration, low blood glucose, insulin resistance and other adverse reactions. We assessed the safety and efficacy of a shortened preoperative fasting period in patients undergoing laparoscopic cholecystectomy. Methods We searched PubMed, Embase and Cochrane Central Register of Controlled Trials up to 20 November 2015 and selected controlled trials with a shortened fasting time before laparoscopic cholecystectomy. We assessed the results by performing a meta-analysis using a variety of outcome measures and investigated the heterogeneity by subgroup analysis. Results Eleven trials were included. Forest plots showed that a shortened fasting time reduced the operative risk and patient discomfort. A shortened fasting time also reduced postoperative nausea and vomiting as well as operative vomiting. With respect to glucose metabolism, a shortened fasting time significantly reduced abnormalities in the ratio of insulin sensitivity. The C-reactive protein concentration was also reduced by a shortened fasting time. Conclusions A shortened preoperative fasting time increases patients' postoperative comfort, improves insulin resistance, and reduces stress responses. This evidence supports the clinical application of a shortened fasting time before laparoscopic cholecystectomy.


Assuntos
Colecistectomia Laparoscópica/efeitos adversos , Desidratação/prevenção & controle , Jejum/efeitos adversos , Hipoglicemia/prevenção & controle , Náusea e Vômito Pós-Operatórios/prevenção & controle , Análise de Variância , Proteína C-Reativa/metabolismo , Colecistectomia Laparoscópica/psicologia , Desidratação/sangue , Desidratação/etiologia , Desidratação/psicologia , Jejum/sangue , Jejum/psicologia , Humanos , Hipoglicemia/sangue , Hipoglicemia/etiologia , Hipoglicemia/psicologia , Resistência à Insulina , Náusea e Vômito Pós-Operatórios/sangue , Náusea e Vômito Pós-Operatórios/etiologia , Náusea e Vômito Pós-Operatórios/psicologia , Cuidados Pré-Operatórios/métodos , Período Pré-Operatório , Estresse Fisiológico , Fatores de Tempo
12.
J Hazard Mater ; 329: 178-184, 2017 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-28135655

RESUMO

The role of industrial byproduct as supplementary cementitious material to partially replace cement has greatly contributed to sustainable environment. Metakaolin (MK), one of such byproduct, is widely used to partial replacement of cement; however, during cement replacement at high percentage, it may not be a good choice to improve the strength of concrete. Thus, in the present study, biocement, a product of microbially induced carbonate precipitation is utilized in MK-modified cement mortars to improve its compressive strength. Despite of cement replacement with MK as high as 50%, the presented technology improved compressive strength of mortars by 27%, which was still comparable to those mortars with 100% cement. The results proved that biomineralization could be effectively used in reducing cement content without compromising compressive strength of mortars. Biocementation also reduced the porosity of mortars at all ages. The process was characterized by SEM-EDS to observe bacterially-induced carbonate crystals and FTIR spectroscopy to predict responsible bonding in the formation of calcium carbonate. Further, XRD analysis identified bio/minerals formed in the MK-modified mortars. The study also encourages combining biological role in construction engineering to solve hazardous nature of cement and at same time solve the disposal problem of industrial waste for sustainable environment.


Assuntos
Carbonato de Cálcio/metabolismo , Materiais de Construção/análise , Força Compressiva , Materiais de Construção/estatística & dados numéricos , Teste de Materiais , Microscopia Eletrônica de Varredura , Porosidade , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios X
13.
Environ Pollut ; 219: 149-155, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27814530

RESUMO

Microbial carbonate precipitation is known as an efficient process for the remediation of heavy metals from contaminated soils. In the present study, a urease positive bacterial isolate, identified as Bacillus cereus NS4 through 16S rDNA sequencing, was utilized on a large scale to remove nickel from industrial soil contaminated by the battery industry. The soil was highly contaminated with an initial total nickel concentration of approximately 900 mg kg-1. The soluble-exchangeable fraction was reduced to 38 mg kg-1 after treatment. The primary objective of metal stabilization was achieved by reducing the bioavailability through immobilizing the nickel in the urease-driven carbonate precipitation. The nickel removal in the soils contributed to the transformation of nickel from mobile species into stable biominerals identified as calcite, vaterite, aragonite and nickelous carbonate when analyzed under XRD. It was proven that during precipitation of calcite, Ni2+ with an ion radius close to Ca2+ was incorporated into the CaCO3 crystal. The biominerals were also characterized by using SEM-EDS to observe the crystal shape and Raman-FTIR spectroscopy to predict responsible bonding during bioremediation with respect to Ni immobilization. The electronic structure and chemical-state information of the detected elements during MICP bioremediation process was studied by XPS. This is the first study in which microbial carbonate precipitation was used for the large-scale remediation of metal-contaminated industrial soil.


Assuntos
Bacillus cereus/metabolismo , Carbonatos/metabolismo , Resíduos Industriais , Níquel/isolamento & purificação , Níquel/metabolismo , Poluentes do Solo/isolamento & purificação , Solo/química , Biodegradação Ambiental , Carbonato de Cálcio/química , Carbonato de Cálcio/metabolismo , Carbonatos/química , Carbonatos/isolamento & purificação , China , Níquel/química , Poluentes do Solo/química , Poluentes do Solo/metabolismo , Solubilidade
14.
Sci Rep ; 6: 32538, 2016 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-27581515

RESUMO

Porcine circovirus type 2 (PCV2) causes porcine circovirus-associated diseases and usually evokes a subclinical infection, without any obvious symptoms, in pigs. It remains unclear how PCV2 leads to a subclinical infection. In this study, we found that peripheral blood mononuclear cells (PBMCs) from PCV2-challenged piglets with no significant clinical symptoms exhibited increased expression of suppressor of cytokine signaling (SOCS) 3, but no significant changes in the expression of the proinflammatory cytokines interleukin (IL)-6 and tumor necrosis factor (TNF)-α; this differed from piglets that displayed significant clinical symptoms. IL-6- and TNF-α-mediated signalings were inhibited in PBMCs from subclinical piglets. Elevated SOCS3 levels inhibited IL-6- and TNF-α-mediated NF-kappa-B inhibitor alpha degradation in PBMCs and PK-15 cells. SOCS3 production was also increased in PCV2-infected PK-15 porcine kidney cells, and IL-6 and TNF-α production that was induced by PCV2 in PK-15 cells was significantly increased when SOCS3 was silenced by a small interfering RNA. SOCS3 interacted with signal transducer and activator of transcription 3 and TNF-associated receptor-associated factor 2, suggesting mechanisms by which SOCS3 inhibits IL-6 and TNF-α signaling. We conclude that SOCS3 plays an important role in PCV2 subclinical infection by suppressing inflammatory responses in primary immune cells.


Assuntos
Infecções por Circoviridae/genética , Circovirus/imunologia , Interleucina-6/genética , Fator de Transcrição STAT3/genética , Proteína 3 Supressora da Sinalização de Citocinas/genética , Fator de Necrose Tumoral alfa/genética , Animais , Infecções Assintomáticas , Linhagem Celular , Infecções por Circoviridae/imunologia , Infecções por Circoviridae/virologia , Circovirus/patogenicidade , Células Epiteliais/imunologia , Células Epiteliais/virologia , Regulação da Expressão Gênica , Interações Hospedeiro-Patógeno/imunologia , Interleucina-6/imunologia , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/virologia , Inibidor de NF-kappaB alfa/genética , Inibidor de NF-kappaB alfa/imunologia , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Fator de Transcrição STAT3/imunologia , Índice de Gravidade de Doença , Transdução de Sinais , Proteína 3 Supressora da Sinalização de Citocinas/antagonistas & inibidores , Proteína 3 Supressora da Sinalização de Citocinas/imunologia , Suínos , Doenças dos Suínos , Fator 2 Associado a Receptor de TNF/genética , Fator 2 Associado a Receptor de TNF/imunologia , Fator de Necrose Tumoral alfa/imunologia
16.
J Surg Res ; 204(2): 288-296, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27565063

RESUMO

BACKGROUND: CXCR3, a G-protein coupled chemokine receptor, has been shown to play a critical role in recruiting inflammatory cells into lungs in several studies. However, its roles in polymicrobial septic acute lung injury (ALI) is yet unknown. Therefore, the purpose of this study was to elucidate the protective effects of CXCR3 blockade on pulmonary microvascular endothelial cells (PMVECs) in septic ALI and explore potential mechanisms. MATERIALS AND METHODS: ALI was induced by polymicrobial sepsis through cecal ligation and puncture surgery. The expression of CXCR3 on pulmonary microvascular endothelial cells was measured 24 h after cecal ligation and puncture surgery. In addition, the protective effects of neutralizing antibody were detected, including protein concentration, inflammation cell counts, lung ​wet-to-dry ratio, and lung damages. In human umbilical vein endothelial cells (HUVECs) culture condition, CXCR3 expression was measured after exposure to tumor necrosis factor-α. The permeability and apoptosis ratio were detected through CXCR3 gene silencing on HUVECs. The p38 mitogen-activated protein kinase (MAPK) was analyzed with Western blot. RESULTS: CXCR3 expression was upregulated both in vivo and in vitro. After CXCR3 neutralizing antibody administrated intraperitoneally, the protein concentration, inflammatory cell counts in BALF and lung ​wet-to-dry ratio were decreased significantly, as well as the lung tissue damages. In vitro, CXCR3 gene silencing inhibited tumor necrosis factor-α and CXCL10-induced hyperpermeability and apoptosis in HUVECs. In addition, p38 mitogen-activated protein kinase activation was essential for CXCR3-mediated apoptosis. CONCLUSIONS: CXCR3 blockade exerts protective effects on ALI at least partly by inhibiting endothelial cells apoptosis and decreasing the leakage of protein-rich fluid and inflammatory cells. Blockade of CXCR3 may be a promising therapeutic strategy for severe sepsis-induced ALI.


Assuntos
Lesão Pulmonar Aguda/metabolismo , Células Endoteliais/metabolismo , Receptores CXCR3/metabolismo , Sepse/complicações , Lesão Pulmonar Aguda/etiologia , Lesão Pulmonar Aguda/patologia , Lesão Pulmonar Aguda/prevenção & controle , Animais , Apoptose , Quimiocina CXCL10/metabolismo , Células Endoteliais da Veia Umbilical Humana , Humanos , Pulmão/patologia , Sistema de Sinalização das MAP Quinases , Masculino , Camundongos Endogâmicos C57BL , Edema Pulmonar/etiologia , Edema Pulmonar/metabolismo , Receptores CXCR3/antagonistas & inibidores , Fator de Necrose Tumoral alfa/metabolismo
17.
J Virol Methods ; 230: 36-44, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26826323

RESUMO

Porcine circovirus type 2 (PCV2) causes significant economic losses to the swine industry worldwide. Heat shock proteins (Hsps) can be used as modulators to enhance both innate and adaptive immune responses. In the present study, recombinant baculoviruses expressing the PCV2Cap protein and the N-terminal 22-370 amino acids of porcine Gp96 (Gp96N), Hsp90, and Hsp70 (rBac-cap/Gp96N, rBac-cap/Hsp90 and rBac-cap/Hsp70, respectively) were constructed and the immune responses were examined in mice and piglets. The mouse experiments showed that rBac-cap/Gp96N increased the titers of specific anti-PCV2 neutralizing antibodies, proliferative responses of peripheral blood mononuclear cells (PBMCs) and IFN-γ levels compared to rBac-cap/Hsp90, rBac-cap/Hsp70, or rBac-cap. The pig experiments showed that the levels of anti-PCV2 antibody, proliferative responses of PBMCs, and IFN-γ in the rBac-cap/Gp96N groups were increased compared to those in rBac-cap group. There were no clear clinical signs of infection following PCV2 challenge in pigs inoculated with recombinant rBac-cap/Gp96N and rBac-cap, and the relative daily weight gains were higher than those in the challenge control (CC) group. The pathological lesions, extent of viremia, and viral loads of the vaccinated groups were milder than those in the CC group. Meanwhile, the extent of viremia and viral load present in the rBac-cap/Gp96N group were significantly lower than those in the rBac-cap group. These results indicated that porcine Gp96N effectively increased the humoral and cell-mediated immune responses of PCV2Cap. Gp96N presents an attractive adjuvant or immunotargeting strategy to enhance the protective efficacy of PCV2 subunit vaccines in swine.


Assuntos
Proteínas do Capsídeo/imunologia , Circovirus/imunologia , Proteínas de Choque Térmico/imunologia , Proteínas Recombinantes/imunologia , Vacinas de Subunidades/imunologia , Vacinas Virais/imunologia , Animais , Baculoviridae , Proteínas do Capsídeo/química , Proteínas de Choque Térmico/química , Camundongos , Proteínas Recombinantes/química , Suínos , Potência de Vacina , Virologia/métodos
18.
Bing Du Xue Bao ; 31(1): 65-73, 2015 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-25997333

RESUMO

Porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) are very two important pathogens that have coursed huge economic losses in swine production in worldwide. In this study,a vector pCMV-TJM containing the full-length cDNA clone of PRRSV attenuated strain TJM-F92 was firstly constructed by PCR method. Then a gene sequence containing Afl II/Mlu I e restriction enzyme sites and a transcription regulatory sequence for ORF6 (TRS6) was inserted be- tween ORF7 and 3'UTR, yielding a expression vector pCMV-TJM-TRS. Subsequently, a plasmid pCMV-TJM-Cap was constructed by cloning of PCV2 ORF2 gene into the unique sites Afl II /Mlu I of pCMV- TJM-TRS plasmid DNA. Then three recombinant PRRSV, rTJM, rTJM/TRS and rTJM/Cap, were rescued by transfection of pCMV-TJM, pCMV-TJM-TRS and pCMV-TJM-Cap into Marc-145 cells, respectively,and confirmed by the genome sequence, restriction enzyme digestion, Western Blot and IFA. They all had the molecular markers which was different from the parent virus. The growth characteristics of the rescued viruses were similar to that of parent virus. rTJM/Cap could also express efficiently PCV2 Cap protein in Marc-145 cells. At passage 8, it still had PCV2 ORF2 gene which examined by RT-PCR. It indicated that the full-length cDNA clone of PRRSV attenuated strain TJM-F92 and recombinant PRRSV rTJM/Cap expressing PCV2 Cap protein were successfully constructed. It made an important foundation for studying on the pathogenic mechanisms of PRRSV and PRRSV-PCV2 vaccine in the future.


Assuntos
Proteínas do Capsídeo/genética , Infecções por Circoviridae/veterinária , Circovirus/genética , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Recombinação Genética , Doenças dos Suínos/virologia , Vacinas Virais/genética , Animais , Proteínas do Capsídeo/imunologia , Linhagem Celular , Infecções por Circoviridae/virologia , Circovirus/classificação , Circovirus/metabolismo , Expressão Gênica , Fases de Leitura Aberta , Síndrome Respiratória e Reprodutiva Suína/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/metabolismo , Suínos , Vacinas Virais/imunologia
19.
RNA Biol ; 12(1): 82-91, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25826415

RESUMO

BACKGROUND: Berberine (BBR) is a natural alkaloid derived from a traditional Chinese herbal medicine. However, the exact mechanisms underlying the different effects of berberine on MM cells have not been fully elucidated. METHODS: A systematic analysis assay integrated common signaling pathways modulated by the 3 miRNA clusters and mRNAs in MM cells after BBR treatment. The role of the mir-99a ∼ 125b cluster, an important oncomir in MM, was identified by comparing the effects of t-anti-mirs with complete complementary antisense locked nucleic acids (LNAs) against mature mir-125b (anti-mir-125b). RESULTS: Three miRNAs clusters (miR-99a ∼ 125b, miR-17 ∼ 92 and miR-106 ∼ 25) were significantly down-regulated in BBR-treated MM cells and are involved in multiple cancer-related signaling pathways. Furthermore, the top 5 differentially regulated genes, RAC1, NFκB1, MYC, JUN and CCND1 might play key roles in the progression of MM. Systematic integration revealed that 3 common signaling pathways (TP53, Erb and MAPK) link the 3 miRNA clusters and the 5 key mRNAs. Meanwhile, both BBR and seed-targeting t-anti-mir-99a ∼ 125b cluster LNAs significantly induced apoptosis, G2-phase cell cycle arrest and colony inhibition. CONCLUSIONS: our results suggest that BBR suppresses multiple myeloma cells, partly by down-regulating the 3 miRNA clusters and many mRNAs, possibly through TP53, Erb and MAPK signaling pathways. The mir-99a ∼ 125b cluster might be a novel target for MM treatment. These findings provide new mechanistic insight into the anticancer effects of certain traditional Chinese herbal medicine compounds.


Assuntos
Alcaloides/farmacologia , Berberina/farmacologia , MicroRNAs/metabolismo , Mieloma Múltiplo/metabolismo , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos , Linhagem Celular Tumoral , Pontos de Checagem da Fase G2 do Ciclo Celular , Humanos , Mieloma Múltiplo/patologia , Oligorribonucleotídeos Antissenso/metabolismo
20.
BMC Syst Biol ; 8: 82, 2014 Jul 07.
Artigo em Inglês | MEDLINE | ID: mdl-25000828

RESUMO

BACKGROUND: Berberine is a natural alkaloid derived from a traditional Chinese herbal medicine. It is known to modulate microRNA (miRNA) levels, although the mechanism for this action is unknown. Here, we previously demonstrate that the expression of 87 miRNAs is differentially affected by berberine in multiple myeloma cells. Among 49 miRNAs that are down-regulated, nine act as oncomirs, including miR-21. Integrative analysis showed that 28 of the down-regulated miRNAs participate in tumor protein p53 (TP53) signaling and other cancer pathways. miR-21 is involved in all these pathways, and is one of the most important oncomirs to be affected by berberine in multiple myeloma cells. RESULTS: We confirmed that berberine down-regulated miRNA-21 expression and significantly up-regulated the expression of programmed cell death 4 (PDCD4), a predicted miR-21 target. Luciferase reporter assays confirmed that PDCD4 was directly regulated by miR-21. Bioinformatic analysis revealed that the miR-21 promoter can be targeted by signal transducer and activator of transcription 3 (STAT3). Down-regulation of interleukin 6 (IL6) by berberine might lead to inhibition of miR-21 transcription through STAT3 down-regulation in multiple myeloma. Furthermore, both berberine and seed-targeting anti-miR-21 oligonucleotide induced apoptosis, G2-phase cell cycle arrest and colony inhibition in multiple myeloma cell lines. Depletion of PDCD4 by short interfering RNA could rescue berberine-induced cytotoxicity in multiple myeloma cells. CONCLUSIONS: Our results suggest that berberine suppresses multiple myeloma cell growth, at least in part, by down-regulating miR-21 levels possibly through IL6/STAT3. This led to increased PDCD4 expression, which is likely to result in suppression of the p53 signaling pathway. These findings may also provide new mechanistic insight into the anti-cancer effects of certain compounds in traditional Chinese herbal medicines.


Assuntos
Antineoplásicos/farmacologia , Berberina/farmacologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , MicroRNAs/genética , Mieloma Múltiplo/patologia , Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose/deficiência , Proteínas Reguladoras de Apoptose/genética , Proteínas Reguladoras de Apoptose/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Biologia Computacional , Regulação para Baixo/efeitos dos fármacos , Pontos de Checagem da Fase G2 do Ciclo Celular/efeitos dos fármacos , Inativação Gênica , Humanos , Interleucina-6/metabolismo , Interleucina-6/farmacologia , Mieloma Múltiplo/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/genética , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Fator de Transcrição STAT3/genética , Regulação para Cima/efeitos dos fármacos
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