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1.
Animals (Basel) ; 14(2)2024 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-38275773

RESUMO

The present study investigates the theoretical basis for maintaining normal physiological functions in heat-stressed beef cattle by exploring the effects of niacin supplementation on the permeability of the rumen epithelial cell barrier. Herein, 12 Jinjiang bulls with an average weight of approximately 400 ± 20.0 kg were randomly divided into three groups, thermoneutral (TN), heat-stressed (HS), and heat-stressed niacin-supplemented (HN) groups, with 4 bulls in each group. The experiment spanned 70 days, and the plasma concentrations of D-lactic acid, diamine oxidase (DAO), lipopolysaccharides (LPSs), and inflammatory cytokines were analyzed. Additionally, we assessed the gene expression of tight junction proteins to understand the effect of niacin supplementation on heat-stressed beef cattle. Our results revealed that heat stress significantly increased the D-lactic acid and LPS levels in beef cattle plasma on days 30 and 45 of the experiment (p < 0.05). Moreover, it led to a significant rise in DAO levels on day 30 (p < 0.05). Niacin supplementation significantly reduced the LPS levels on day 30 (p < 0.05). Heat stress significantly elevated the plasma concentrations of inflammatory cytokines interleukin-1ß (IL-1ß), IL-2, IL-6, and tumor necrosis factor-α (TNF-α) (p < 0.05), while reducing the IL-4 concentration (p < 0.05). However, niacin supplementation effectively mitigated the concentrations of these inflammatory factors by reducing IL-1ß, IL-2, IL-6, and TNF-α concentrations and increasing IL-4 concentrations. The mRNA expressions of tight junction proteins zonula occluden-1 (ZO-1), claudin-1, claudin-4, and claudin-7 were significantly downregulated (p < 0.05) in the HS group compared to those in the TN group, and those of ZO-1 and occludin were significantly upregulated (p < 0.05) in the HN group compared to those in the HS group. Notably, no significant differences were observed in ruminal papillae length and width among the studied groups (p > 0.05). Our findings indicate that heat stress adversely impacted the tight junction structure of the rumen epithelium, leading to a significant reduction in the expression of tight junction protein mRNA. Consequently, heat stress impaired the rumen mucosal barrier function, resulting in increased intestinal permeability. The mechanism underlying this effect may be associated with the decreased expression of tight junction protein genes in the rumen epithelial cells. However, niacin supplementation mitigated the detrimental effects of heat stress on intestinal permeability in beef cattle and increased the expression of tight junction protein genes in the rumen epithelium, thereby effectively protecting the rumen barrier in heat-stressed beef cattle. These results highlight the potential of nicotinic acid as a protective agent against the negative impacts of heat stress on intestinal integrity in beef cattle.

2.
J Cereb Blood Flow Metab ; 44(1): 105-117, 2024 01.
Artigo em Inglês | MEDLINE | ID: mdl-37717175

RESUMO

Cerebrospinal fluid (CSF) flow patterns and their relationship with arterial pulsation can depict the function of glymphatic system (GS). We propose an improved multi-directional diffusion-sensitized driven-equilibrium (iMDDSDE) prepared heavily T2-weighted 3D FSE (iMDDSDE-HT2) magnetic resonance imaging (MRI) method to noninvasively assess the mobility (MO) of CSF distributed in the ventricles and perivascular spaces (PVS). This method could obtain 3D high resolution (1 mm isotropic) imaging of CSF MO with full brain coverage within five min and distinguish the CSF MO across different pulse phases using a peripheral pulse unit (PPU). The MO curves had the largest amplitude value in the PVS of middle cerebral artery (11.11 × 10-9 m2/s) and the largest amplitude growth rate in the posterior cerebral artery (189%). The average coefficient of variations (CVs) in non-pulse trigger and pulse phase 1 and 3 were 0.11, 0.10 and 0.09 respectively. The MO in older healthy participants was lower compared to the young participants, and the MO in cerebral major artery stenosis patients with acute ischemia stroke (AIS) were lower compared to those without AIS in several ventriclar ROIs (P < 0.05). This sequence is a clinically feasible method to effectively evaluate CSF flow patterns in human brain.


Assuntos
Encéfalo , Imageamento por Ressonância Magnética , Humanos , Idoso , Imageamento por Ressonância Magnética/métodos , Cabeça , Líquido Cefalorraquidiano/diagnóstico por imagem , Imageamento Tridimensional/métodos
3.
Med ; 4(12): 898-912.e4, 2023 Dec 08.
Artigo em Inglês | MEDLINE | ID: mdl-37944532

RESUMO

BACKGROUND: Meningeal lymphatic vessels (mLVs) have proven to bear a relationship with tumor immunity and therapeutic efficacy of intracranial malignant tumors in pre-clinical animal studies. We aimed to explore the association between mLV function and intracranial malignant tumors in clinical participants. METHODS: The participants were allocated to a control group or a group of patients with intracranial tumors. Dynamic enhanced magnetic resonance was used to evaluate the wash-in and wash-out functions of mLVs around the superior sagittal sinus and the sigmoid sinus. FINDINGS: A total of 246 individuals were recruited for our study. The area under curve and wash-in rate of mLVs in the intracranial tumor group were higher than in the control group (2,749 vs. 2,110, p < 0.001 and 3.72 vs. 2.87, p < 0.001, respectively). The wash-out ratio of mLVs was lower in the intracranial tumor group than in the control group (0.65 vs. 0.73, p < 0.001). Decreased wash-out of mLVs was associated with tumor progression (ß = -0.118; p < 0.001). High-grade glioma and isocitrate dehydrogenase wild type were associated with a lower mLV wash-out function (ß = -0.057, p = 0.044 and ß = -0.069, p = 0.047, respectively). CONCLUSIONS: Intracranial malignant tumors were associated with elevated wash-in function and decreased wash-out function of mLVs. High-grade glioma and isocitrate dehydrogenase wild type were associated with low mLV wash-out function, and long-term decreased mLV wash-out function was a risk factor for tumor progression. FUNDING: There was no funding.


Assuntos
Neoplasias Encefálicas , Glioma , Vasos Linfáticos , Animais , Humanos , Isocitrato Desidrogenase , Neoplasias Encefálicas/diagnóstico por imagem , Imageamento por Ressonância Magnética
4.
Opt Express ; 20(16): 17741-59, 2012 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-23038326

RESUMO

Nowadays, there is a hot debate among industry and academic researchers that whether the newly developed scientific-grade Complementary Metal Oxide Semiconductor (sCMOS) cameras could become the image sensors of choice in localization-based super-resolution microscopy. To help researchers find answers to this question, here we reported an experimental methodology for quantitatively comparing the performance of low-light cameras in single molecule detection (characterized via image SNR) and localization (via localization accuracy). We found that a newly launched sCMOS camera can present superior imaging performance than a popular Electron Multiplying Charge Coupled Device (EMCCD) camera in a signal range (15-12000 photon/pixel) more than enough for typical localization-based super-resolution microscopy.


Assuntos
Elétrons , Metais/química , Microscopia/instrumentação , Microscopia/métodos , Óxidos/química , Fotografação/instrumentação , Semicondutores , Fluorescência , Processamento de Imagem Assistida por Computador , Fótons , Razão Sinal-Ruído
5.
Opt Lett ; 37(13): 2481-3, 2012 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-22743428

RESUMO

We present an algorithm to estimate the location of single fluorescent molecule with both high speed and high precision. This algorithm is based on finding the subpixel position with maximum radial symmetry in a pixelated single molecule fluorescence image. Compared with conventional algorithms, this algorithm does not rely on point-spread-function or noise model. Through numerical simulation and experimental analysis, we found that this algorithm exhibits localization precision very close to the maximum likelihood estimator (MLE), while executes ∼1000 times faster than the MLE and ∼6 times faster than the fluoroBancroft algorithm.


Assuntos
Algoritmos , Espectrometria de Fluorescência/métodos , Corantes Fluorescentes , Processamento de Imagem Assistida por Computador , Fatores de Tempo
6.
Front Microbiol ; 13: 975346, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36274720

RESUMO

The objective of this study was to investigate the alleviation effects of niacin supplementation on beef cattle subjected to heat stress and to provide a theoretical basis for exploring the alleviation methods of heat stress environmental factors on the rumen of beef cattle. In the experiment, 36 Jinjiang bull cattle with a body weight of about 400 ± 20.0 kg were randomly divided into three treatments, each treatment contains four replicates, with three cattle in each replicate. Treatments included thermoneutral treatment (TN; temperature: 24-25°C, humidity: 45-55%), heat stress treatment, exposure to environmental temperature (HS; average THI: 82.74), and heat stress supplemented with niacin treatment (HN; high temperature + 800 mg/kg NA). Measured indicators were body temperature, respiratory rate, production performances, rumen fermentations, and microbial diversity. Results showed that adding niacin reduced the body temperature and respiratory rate (P < 0.05) but had no significant effect on the production performances compared with heat-stressed beef cattle. HS treatment significantly increased body temperature and respiratory rate (P < 0.01), while decreasing the content of acetic acid, butyric acid, and total volatile fatty acids (P < 0.05) compared with the TN treatment. Supplement of niacin did not affect ruminal fermentation parameters (P > 0.05) but had a decreased tendency on A/P (P < 0.1). Microbial diversity results showed that, at the phylum level, the relative abundance of Desulfobacterota in the HS treatment was increased compared with TN and HN treatment (P < 0.05). At the genus level, the relative abundance of Succiniclasticum and Family_XIII_AD3011 group in the HN treatment significantly proliferated compared with the HS treatment (P < 0.05). In conclusion, niacin supplementation may alleviate heat stress by proliferating those bacteria belonging to the phylum Succiniclasticum, which may further contribute to the digestion of cellulose and the improvement of the metabolic function of Jinjiang cattle under heat-stress conditions.

7.
Opt Express ; 19(20): 19156-68, 2011 Sep 26.
Artigo em Inglês | MEDLINE | ID: mdl-21996858

RESUMO

In the community of localization-based super-resolution microscopy (or called localization microscopy), it is generally believed that the emission of single molecules is so weak that an EMCCD (electron multiplying charge coupled device) camera is necessary to be used as the detector by eliminating read noise. Here we evaluate the possibility of a new kind of low light detector, scientific complementary metal-oxide-semiconductor (sCMOS) camera in localization microscopy. We demonstrate experimentally that sCMOS is capable of imaging actin bundles with FWHM diameter of 37 nm, evidencing the capability of sCMOS in localization microscopy. We further characterize the noise performance of sCMOS and find out that, with the use of a bright fluorescence probe such as d2EosFP, localization microscopy imaging is now working in the shot noise limited region.


Assuntos
Microscopia/instrumentação , Fótons , Semicondutores , Processamento de Sinais Assistido por Computador/instrumentação , Desenho de Equipamento
8.
Front Immunol ; 12: 675534, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34335573

RESUMO

The RNA-binding protein tristetraprolin (TTP) is an anti-inflammatory factor that prompts the mRNA decay of target mRNAs and is involved in inflammatory diseases such as rheumatoid arthritis (RA). TTP is regulated by phosphorylation, and protein phosphatase 2A (PP2A) can dephosphorylate TTP to activate its mRNA-degrading function. Some small molecules can enhance PP2A activation. Short interfering RNA (siRNA) targeting TTP expression or PP2A agonist (Arctigenin) was administered to monosodium urate (MSU) crystal-induced J774A.1 cells, and the expression of inflammatory related genes was detected by RT-PCR and Western blot assays. The effects of Arctigenin in mouse models of acute inflammation induced by MSU crystals, including peritonitis and arthritis, were evaluated. The data indicated that TTP expression levels and endogenous PP2A activity were increased in MSU-crystal treated J774A.1 cells. TTP knockdown exacerbated inflammation-related genes expression and NLRP3 inflammasome activation. However, PP2A agonist treatment (Arctigenin) suppressed MSU crystal-induced inflammation in J774A.1 cells. Arctigenin also relieved mitochondrial reactive oxygen species (mtROS) production and improved lysosomal membrane permeability in MSU crystal-treated J774A.1 cells. Moreover, TTP knockdown reversed the anti-inflammatory and antioxidant effects of Arctigenin. Oral administration of Arctigenin significantly alleviated foot pad swelling, the number of inflammatory cells in peritoneal lavage fluids and the production of IL-1ß in the mouse model of inflammation induced by MSU crystals. Collectively, these data imply that targeting TTP expression or functional activity may provide a potential therapeutic strategy for inflammation caused by MSU crystals.


Assuntos
Furanos/farmacologia , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Lignanas/farmacologia , Tristetraprolina/genética , Tristetraprolina/metabolismo , Tristetraprolina/fisiologia , Animais , Anti-Inflamatórios/farmacologia , Autofagia , Caspase 1/metabolismo , Técnicas de Cultura de Células , Citocinas/metabolismo , Lisossomos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Mitocôndrias/metabolismo , Proteína Fosfatase 2/metabolismo , RNA Interferente Pequeno , Espécies Reativas de Oxigênio/metabolismo , Ácido Úrico/farmacologia
9.
Opt Express ; 18(11): 11867-76, 2010 May 24.
Artigo em Inglês | MEDLINE | ID: mdl-20589048

RESUMO

Localization-based super resolution microscopy holds superior performances in live cell imaging, but its widespread use is thus far mainly hindered by the slow image analysis speed. Here we show a powerful image analysis method based on the combination of the maximum likelihood algorithm and a Graphics Processing Unit (GPU). Results indicate that our method is fast enough for real-time processing of experimental images even from fast EMCCD cameras working at full frame rate without compromising localization precision or field of view. This newly developed method is also capable of revealing movements from the images immediately after data acquisition, which is of great benefit to live cell imaging.


Assuntos
Algoritmos , Aumento da Imagem/instrumentação , Aumento da Imagem/métodos , Microscopia/instrumentação , Microscopia/métodos , Processamento de Sinais Assistido por Computador/instrumentação , Desenho de Equipamento , Análise de Falha de Equipamento , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
10.
Oxid Med Cell Longev ; 2020: 8706898, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33488933

RESUMO

Acute gout is an inflammatory response induced by monosodium urate (MSU) crystals. HSP60 is a highly conserved stress protein that acts as a cellular "danger" signal for immune reactions. In this study, we aimed to investigate the role and molecular mechanism of HSP60 in gout. HSP60 expression was detected in peripheral blood mononuclear cells (PBMCs) and plasma of gout patients. The effect and molecular mechanism of HSP60 in gout were studied in MSU crystals treatment macrophages and C57BL/6 mice. JC-1 probe and MitoSOX Red were used to measure the mitochondrial membrane potential (MMP) and mitochondrial reactive oxygen species (mtROS). HSP60 expression was significantly upregulated in the PBMCs and sera of patients with acute gout (AG) compared to those with intercritical gout (IG) or healthy controls (HCs). MSU crystals induced the expression and secretion of HSP60 in the macrophages. HSP60 knockdown or overexpression affects TLR4 and MyD88 expression, IκBα degradation, and the nuclear localization of NF-κB in MSU crystal-stimulated inflammation. Further, HSP60 facilitates MMP collapse and mtROS production and activates the NLRP3 inflammasome in MSU crystal-stimulated macrophages. In MSU crystal-induced arthritis mouse models pretreated with HSP60 vivo-morpholino, paw swelling, myeloperoxidase (MPO) activity, and inflammatory cell infiltration significantly decreased. Our study reveals that MSU crystal stimulates the expression of HSP60, which accelerates the TLR4-MyD88-NF-κB signaling pathway and exacerbates mitochondrial dysfunction.


Assuntos
Artrite Experimental/patologia , Chaperonina 60/metabolismo , Gota/patologia , Inflamação/patologia , Leucócitos Mononucleares/patologia , Mitocôndrias/patologia , Ácido Úrico/toxicidade , Adulto , Animais , Antioxidantes/toxicidade , Artrite Experimental/induzido quimicamente , Artrite Experimental/imunologia , Artrite Experimental/metabolismo , Estudos de Casos e Controles , Chaperonina 60/genética , Gota/etiologia , Gota/metabolismo , Humanos , Inflamassomos , Inflamação/induzido quimicamente , Inflamação/imunologia , Inflamação/metabolismo , Leucócitos Mononucleares/metabolismo , Macrófagos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Pessoa de Meia-Idade , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Fator 88 de Diferenciação Mieloide/genética , Fator 88 de Diferenciação Mieloide/metabolismo , NF-kappa B/genética , NF-kappa B/metabolismo , Transdução de Sinais , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/metabolismo
11.
J Environ Sci (China) ; 18(1): 62-8, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-20050550

RESUMO

With the development of activated sludge model, the simulation software for the design and operation of wastewater treatment plant (WWTP) was produced and has been widely used. The dynamic change of the quality and flow of influent are major factors causing the unstable operation of wastewater treatment process. As a basic model, ASM1 model was used for the simulation of activated sludge process, and double exponential model was selected for the simulation of secondary sedimentation tank. The influences of influent change to the aeration tank and secondary sedimentation tank were investigated, and the relationship among influent change, the quality of effluent and the level of sludge blanket in secondary sedimentation tank was established. On the basis of the simulation results, the operation of the WWTP could be adjusted under the dynamic change of the influent. Furthermore, the controlling strategy combined the feed-forward on the influent flow and the feedback on the level of sludge blanket in the secondary sedimentation tank was studied.


Assuntos
Poluentes da Água/isolamento & purificação , Modelos Teóricos
12.
BMC Genomics ; 5(1): 26, 2004 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-15113399

RESUMO

BACKGROUND: Several high throughput technologies have been employed to identify differentially regulated genes that may be molecular targets for drug discovery. Here we compared the sets of differentially regulated genes discovered using two experimental approaches: a subtracted suppressive hybridization (SSH) cDNA library methodology and Affymetrix GeneChip technology. In this "case study" we explored the transcriptional pattern changes during the in vitro differentiation of human monocytes to myeloid dendritic cells (DC), and evaluated the potential for novel gene discovery using the SSH methodology. RESULTS: The same RNA samples isolated from peripheral blood monocyte precursors and immature DC (iDC) were used for GeneChip microarray probing and SSH cDNA library construction. 10,000 clones from each of the two-way SSH libraries (iDC-monocytes and monocytes-iDC) were picked for sequencing. About 2000 transcripts were identified for each library from 8000 successful sequences. Only 70% to 75% of these transcripts were represented on the U95 series GeneChip microarrays, implying that 25% to 30% of these transcripts might not have been identified in a study based only on GeneChip microarrays. In addition, about 10% of these transcripts appeared to be "novel", although these have not yet been closely examined. Among the transcripts that are also represented on the chips, about a third were concordantly discovered as differentially regulated between iDC and monocytes by GeneChip microarray transcript profiling. The remaining two thirds were either not inferred as differentially regulated from GeneChip microarray data, or were called differentially regulated but in the opposite direction. This underscores the importance both of generating reciprocal pairs of SSH libraries, and of real-time RT-PCR confirmation of the results. CONCLUSIONS: This study suggests that SSH could be used as an alternative and complementary transcript profiling tool to GeneChip microarrays, especially in identifying novel genes and transcripts of low abundance.


Assuntos
Células Dendríticas/metabolismo , Perfilação da Expressão Gênica/métodos , Monócitos/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Biblioteca Gênica , Humanos , Hibridização de Ácido Nucleico/métodos , RNA/genética , RNA/metabolismo , Reprodutibilidade dos Testes
13.
Artigo em Inglês | MEDLINE | ID: mdl-12237682

RESUMO

In order to enhance the thermostability of D-Glucose isomerase (GI), its mutants, GIK253R and GIN184V, were obtained with the double primer method of site-directed mutagenesis. Then their biochemical properties were assayed and compared with wild-type GI. The results showed that: (1) The mutant K253R was less stable than the wild-type GI at 70 degrees and 80 degrees. But in 1 M L-Rhaminose at 70 degrees, they had similar rate of heat inactivation. Furthermore, the mutant K253R has higher specific activity than the wild-type GI. (2) The mutant N184V had much less thermostability and specific activity as compared with the wild-type GI. These results were explained by their kinetic parameters and crystal structure model.

14.
Eur J Immunol ; 35(9): 2709-17, 2005 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-16106470

RESUMO

Upon activation in vitro, only a fraction of the bulk human T helper cell cultures secret the hallmark Th1/2 cytokines (IFN-gamma for Th1 and IL-4 for Th2, respectively). It is uncertain whether these IFN-gamma-/IL-4- cells are differentiated Th1 or Th2 cells. Here, we have characterized live IFN-gamma+, IL-4+ and IFN-gamma-/IL-4- cells isolated from Th cell cultures treated under Th1 or Th2 polarizing conditions by employing affinity matrix capture technology. RNA samples from the sorted cells were analyzed by real time RT-PCR and microarrays. The double negative cells from either Th1 or Th2 cultures expressed lower levels of Th1/Th2 marker cytokine genes (IFNgamma, IL4, and IL5). However, they were comparable with the IFN-gamma+ or IL-4+ cells in the expression levels of other Th1/Th2 marker genes (GATA3, Tbet, and IL12Rbeta2). Most importantly, these double negative cells were already committed in their Th1/Th2 lineages. Gene expression profiling analysis showed that very few previously identified Th1/Th2 marker genes were differentially expressed between the IFN-gamma or IL-4 producers and the non-producers, further underscoring the similarity between these two groups.


Assuntos
Interferon gama/metabolismo , Interleucina-4/metabolismo , Células Th1/imunologia , Células Th2/imunologia , Diferenciação Celular/imunologia , Linhagem da Célula/imunologia , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/imunologia , Citometria de Fluxo , Fator de Transcrição GATA3 , Expressão Gênica , Marcadores Genéticos/imunologia , Humanos , Interferon gama/genética , Interferon gama/imunologia , Interleucina-4/genética , Interleucina-4/imunologia , RNA/química , RNA/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células Th1/citologia , Células Th1/metabolismo , Células Th2/citologia , Células Th2/metabolismo , Transativadores/genética , Transativadores/imunologia
15.
In Silico Biol ; 4(4): 395-410, 2004.
Artigo em Inglês | MEDLINE | ID: mdl-15506990

RESUMO

DNA-binding transcription factors play a central role in transcription regulation, and the annotation of transcription-factor binding sites in upstream regions of human genes is essential for building a genome-wide regulatory network. We describe methodology to accurately predict the transcription-factor binding sites in the proximal-promoter region of function-specific genes. In order to increase the accuracy of transcription factor binding-site prediction, we rely on recent genome sequence data, known transcription factor binding-site matrices, and Gene Ontology biological-function-based gene classification. Using TRANSFAC position-frequency matrices, we detected individual and cooperating transcription-factor binding sites in proximal promoters of ENSEMBL annotated human genes. We used the over representation of detected binding sites in the proximal promoters as compared to the second exons to control specificity. We confirmed the majority of transcription-factor binding sites predicted in proximal promoters of immune-response genes with evidence from existing literature. We validated the predicted cooperation between transcription factors NF-kappa B and IRF in the regulation of gene expression with microarray transcript profiling data and literature-derived protein-protein interaction network. We also identified over-represented individual and pairs of transcription-factor binding sites in the proximal promoters of each Gene Ontology biological-process gene group. Our tools and analysis provide a new resource for deciphering transcription regulation in different biological paradigms.


Assuntos
Biologia Computacional/métodos , Proteínas de Ligação a DNA/metabolismo , Genoma Humano , Genômica/métodos , Regiões Promotoras Genéticas/genética , Fatores de Transcrição/metabolismo , Éxons/genética , Genes MHC da Classe II/genética , Humanos , NF-kappa B/genética , NF-kappa B/fisiologia
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