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1.
World J Clin Cases ; 10(1): 205-216, 2022 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-35071519

RESUMO

BACKGROUND: Herein, we report the genetic, clinical, molecular and biochemical features of two Han Chinese pedigrees with suggested maternally transmitted non-syndromic hearing loss. AIM: To investigate the pathophysiology of hearing loss associated with mitochondrial tRNA mutations. METHODS: Sixteen subjects from two Chinese families with hearing loss underwent clinical, genetic, molecular, and biochemical evaluations. Biochemical characterizations included the measurements of tRNA levels using lymphoblastoid cell lines derived from five affected matrilineal relatives of these families and three control subjects. RESULTS: Three of the 16 matrilineal relatives in these families exhibited a variable seriousness and age-at-onset (8 years) of deafness. Analysis of mtDNA mutation identified the novel homoplasmic tRNAIle 4268T>C mutation in two families both belonging to haplogroup D4j. The 4268T>C mutation is located in a highly conserved base pairing (6U-67A) of tRNAIle. The elimination of 6U-67A base-pairing may change the tRNAIle metabolism. Functional mutation was supported by an approximately 64.6% reduction in the level of tRNAIle observed in the lymphoblastoid cell lines with the 4268T>C mutation, in contrast to the wild-type cell lines. The reduced level of tRNA was below the proposed threshold for normal respiration in lymphoblastoid cells. However, genotyping analysis did not detect any mutations in the prominent deafness-causing gene GJB2 in any members of the family. CONCLUSION: These data show that the novel tRNAIle 4268T>C mutation was involved in maternally transmitted deafness. However, epigenetic, other genetic, or environmental factors may be attributed to the phenotypic variability. These findings will be useful for understanding families with maternally inherited deafness.

2.
Comput Struct Biotechnol J ; 19: 4042-4048, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34527183

RESUMO

Studies on codon property would deepen our understanding of the origin of primitive life and enlighten biotechnical application. Here, we proposed a quantitative measurement of codon-amino acid association and found that seven out of 13 physicochemical properties have stronger associations with the nucleotide identity at the second codon position, indicating that protein structure and function may associate more closely with it than the other two sites. When extending the effect of codon-amino acid association to protein level, it was found that the correlation between the second codon position (measured by the relative frequencies of nucleobase T and A at this codon site) and hydrophobicity (by the form of GRAVY value) became stronger with 96% genomes having R > 0.90 and p < 1e-60. Furthermore, we revealed that informational genes encoding proteins have lower GRAVY values than operational proteins (p < 3e-37) in both prokaryotic and eukaryotic genomes. The above results reveal a complete link from codon identity (A2 versus T2) to amino acid property (hydrophilic versus hydrophobic) and then to protein functions (informational versus operational). Hence, our work may help to understand how the nucleotide sequence determines protein function.

3.
BMC Genomics ; 22(1): 561, 2021 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-34289810

RESUMO

BACKGROUND: The B3 superfamily (B3s) represents a class of large plant-specific transcription factors, which play diverse roles in plant growth and development process including flowering induction. However, identification and functional surveys of B3 superfamily have not been reported in ethylene-induced pineapple flowering (Ananas comosus). RESULTS: 57 B3 genes containing B3 domain were identified and phylogenetically classified into five subfamilies. Chromosomal localization analysis revealed that 54 of 57 AcB3s were located on 21 Linkage Groups (LG). Collinearity analysis demonstrated that the segmental duplication was the main event in the evolution of B3 gene superfamily, and most of them were under purifying selection. The analysis of cis-element composition suggested that most of these genes may have function in response to abscisic acid, ethylene, MeJA, light, and abiotic stress. qRT-PCR analysis of 40 AcB3s containing ethylene responsive elements exhibited that the expression levels of 35 genes were up-regulated within 1 d after ethephon treatment and some were highly expressed in flower bud differentiation period in stem apex, such as Aco012003, Aco019552 and Aco014401. CONCLUSION: This study provides a basic information of AcB3s and clues for involvement of some AcB3s in ethylene-induced flowering in pineapple.


Assuntos
Ananas , Ananas/genética , Etilenos , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genoma de Planta , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
4.
Oncol Lett ; 17(6): 5536-5544, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31186775

RESUMO

Head and neck squamous cell carcinoma (HNSC) is one of most common types of cancer worldwide, and mRNAs and long non-coding RNAs (lncRNAs) have been identified as prognostic biomarkers in HNSC. In the present study, using gene expression datasets from multiple platforms, survival-associated genes in HNSC were identified. Subsequently, a combination of 17 genes (14 mRNAs and 3 lncRNA) was optimized using random forest variable hunting and a risk score model for HNSC prognosis was developed using a cohort from The Cancer Genome Atlas. Patients with high-risk scores tend to have earlier disease recurrence and lower survival rates, compared with those with low-risk scores. This observation was further validated in three independent datasets (GSE41613, GSE10300 and E-MTAB-302). Association analysis revealed that the risk score is independent of other clinicopathological observations. On the basis of the results depicted in the nomogram, the risk score performs better in 3-year survival rate prediction than other clinical observations. In summary, the lncRNA-mRNA signature-based risk score successfully predicts the survival of HNSC and serves as an indicator of prognosis.

5.
Ecotoxicol Environ Saf ; 176: 234-241, 2019 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-30939403

RESUMO

The present study aimed to explore the repair effect and mechanism of bone marrow mesenchymal stem cells (BMSCs) transplantation on injured kidneys caused by hexavalent chromium (Cr (VI)). Wistar rats were intraperitoneally injected with 0.4 mg/kg•bw Cr (VI) ion solution. After 30 days, 1 × 107 BMSCs were transplanted into rats. After cell transplantation for 2 weeks, there was no significant difference in the chromium content between the model and BMSCs-therapy group by atomic absorption spectrometry. In BMSCs-therapy group, the renal organ index, the serum levels of blood urea nitrogen (BUN) and creatinine (CRE), malonaldehyde (MDA) content were significantly decreased, superoxide dismutase (SOD) activity was significantly elevated, and the pathological changes were improved compared with the model group. The results of immunohistochemical and western blot assays showed that the expressions of apoptosis-related proteins Bax, Cytochrome c, and Caspase-3, as well as autophagy-associated proteins Beclin 1, PINK1, Parkin, p-Parkin, LC3B, and the MAPK signaling pathway, including the ratio of p-p38 to p38 and p-JNK to JNK were all significantly decreased, Bcl-2 and p62 expressions, and the ratio of p-ERK to ERK were significantly elevated in BMSCs-therapy group compared with the model group. These results suggested that BMSCs repaired Cr (VI)-injured kidney through decreasing mitochondria-mediated apoptosis and mitophagy mediated by downregulating phosphorylation of p38 and JNK, upregulating phosphorylation of ERK.


Assuntos
Apoptose/efeitos dos fármacos , Cromo/toxicidade , Nefropatias/terapia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Transplante de Células-Tronco Mesenquimais , Mitofagia/efeitos dos fármacos , Animais , Autofagia/efeitos dos fármacos , Rim/efeitos dos fármacos , Rim/metabolismo , Rim/patologia , Nefropatias/metabolismo , Nefropatias/patologia , Testes de Função Renal , Masculino , Fosforilação , Ratos , Ratos Wistar
6.
Toxicol Lett ; 310: 31-38, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30974164

RESUMO

This study focused on the effect of bone marrow mesenchymal stem cells (BMSCs) on the repair of rat liver injury induced by Cr (VI). Twenty-four Wistar rats were randomly divided into the control, model and cell therapy group, with 8 rats in each group. Potassium dichromate solution containing 0, 0.4 and 0.4 mg/kg·bw Cr (VI) was administered 5 times a week for 30 days. At the end of treatment, rats in the cell therapy group were administered 1 × 107 BMSCs. Two weeks later, serum alanine and aspartate aminotransferase levels in the cell therapy group were significantly improved compared with those in the model group, CM-Dil-labeled BMSCs were localized in rat livers. Compared with the model group, in the cell therapy group the number of apoptotic hepatocytes by TUNEL assay, MDA content, the expression of HIF-1α, endoplasmic reticulum (ER) stress-mediated apoptosis-related proteins including Grp78, CHOP, Cleaved-Caspase-12, ATF6, and Bax was significantly lower, and SOD activity, the expression of SIRT1 and Bcl-2 was significantly higher. It is suggested that BMSCs are localized in livers and reduce the toxic effects of Cr (VI) on the liver, and the possible mechanism may be related to the mechanisms of BMSCs decreasing ER stress-mediated hepatocyte apoptosis via the SIRT1/HIF-1α signaling pathway.


Assuntos
Apoptose/efeitos dos fármacos , Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Cromo/toxicidade , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Fígado/efeitos dos fármacos , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/enzimologia , Transdução de Sinais/efeitos dos fármacos , Sirtuína 1/metabolismo , Animais , Comunicação Celular , Diferenciação Celular , Células Cultivadas , Doença Hepática Induzida por Substâncias e Drogas/enzimologia , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Doença Hepática Induzida por Substâncias e Drogas/patologia , Fígado/enzimologia , Fígado/patologia , Ratos Wistar
7.
Sci Rep ; 7(1): 309, 2017 03 22.
Artigo em Inglês | MEDLINE | ID: mdl-28331188

RESUMO

Dysregulation of mRNAs and long non-coding RNAs (lncRNAs) is one of the most important features of carcinogenesis and cancer development. However, studies integrating the expression of mRNAs and lncRNAs to predict the survival of head and neck squamous cell carcinoma (HNSC) are still limited, hitherto. In current work, we identified survival related mRNAs and lncRNAs in three datasets (TCGA dataset, E-TABM-302, GSE41613). By random forest, seven gene signatures (six mRNAs and lncRNA) were further selected to develop the risk score model. The risk score was significantly associated with survival in both training and testing datasets (E-TABM-302, GSE41613, and E-MTAB-1324). Furthermore, correlation analyses showed that the risk score is independent from clinicopathological features. According to Cox multivariable hazard model and nomogram, the risk score contributes the most to survival than the other clinical information, including gender, age, histologic grade, and alcohol taking. The Gene Set Enrichment Analysis (GSEA) indicates that the risk score is associated with cancer related pathways. In summary, the lncRNA-mRNA based risk score model we developed successfully predicts the survival of 755 HNSC samples in five datasets and two platforms. It is independent from clinical information and performs better than clinical information for prognosis.


Assuntos
Carcinoma de Células Escamosas/diagnóstico , Carcinoma de Células Escamosas/patologia , Neoplasias de Cabeça e Pescoço/diagnóstico , Neoplasias de Cabeça e Pescoço/patologia , RNA Longo não Codificante/análise , RNA Mensageiro/análise , Carcinoma de Células Escamosas/mortalidade , Neoplasias de Cabeça e Pescoço/mortalidade , Humanos , Prognóstico , Medição de Risco , Análise de Sobrevida
8.
Zhongguo Zhong Yao Za Zhi ; 41(22): 4169-4174, 2016 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-28933084

RESUMO

The open reading frame of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (HDR) was cloned from Phlegmarirus carinatus by RT-PCR method and the sequence was analyzed by bioinformatics tools. After searching the transcriptome dataset of P. carinatus, one unique sequence encoding 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase was discovered. The primers were designed according to the cDNA sequence of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase from the dataset. And then, the open reading frame (ORF) of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase, named as PcHDR1 (GenBank Accession number:JQ957845), was cloned by RT-PCR strategy with the template of mixed RNA extracted from roots, stem and leaf of P. carinatus. The bioinformatic analysis of this gene and its corresponding protein was performed. The ORF of PcHDR1 consisted of 1 437 base pairs (bp), encoding one polypeptide with 478 amino acids. The sequence comparison showed that PcHDR1 is closest with GbHDR (Ginkgo biloba),and the sequence homology was up to 78%. Bioinformatics prediction and analysis indicated that PcHDR1 protein contained a conserved domain of LytB, without transmembrane region and signal peptides. This study cloned and analyzed 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase from P. carinatus. The result will provide a foundation for exploring the function of PcHDR1 involved in terpene biosynthesis in P. carinatus plants.


Assuntos
Lycopodiaceae/enzimologia , Lycopodiaceae/genética , Oxirredutases/genética , Sequência de Aminoácidos , Clonagem Molecular , Biologia Computacional , DNA Complementar , Genes de Plantas , Filogenia
9.
Beijing Da Xue Xue Bao Yi Xue Ban ; 47(5): 838-41, 2015 Oct 18.
Artigo em Chinês | MEDLINE | ID: mdl-26474627

RESUMO

OBJECTIVE: To find the best synthesis method of 6-benzyl-1-[(benzyloxy)methyl]-3-hydroxy-5-(hydroxymethyl) pyrimidine-2,4(1H,3H)-dione e for observing the change of its biological activity after N-3 hydroxylation. METHODS: After trying some N-hydroxylation methods, the target compound was successfully synthesized via one-pot oxidizing process by sodium hydride (NaH) and 3-chloroperbenzoic acid(m-CPBA); the anti-HIV reverse transcriptase (RT) activity and integrase (IN) activity of the target compound was assayed via enzyme-linked immunesorbent assay (ELISA) and phosphorylation of DNA package method. RESULTS: The target compound could be obtained through the improved m-CPBA oxidative method by only one step, and the yield of the reaction could reach 60%-70%. And the structure of this compound was identified by 1H NMR, 13C NMR and MS; The activity result showed it added the anti-HIV IN activity after N-3 hydroxylation as well as retained the anti-HIV RT activity. CONCLUSION: The improved m-CPBA oxidative method is a convenient and efficient way to prepare the compound 6-benzyl-1-[(benzyloxy)methyl]-3-hydroxy-5-(hydroxymethyl)pyrimidine-2,4(1H,3H)-dione e which has both anti-HIV RT and IN activity.


Assuntos
Fármacos Anti-HIV/síntese química , Inibidores de Integrase de HIV/síntese química , Transcriptase Reversa do HIV/antagonistas & inibidores , HIV-1/efeitos dos fármacos , Nucleosídeos de Pirimidina/síntese química
10.
Beijing Da Xue Xue Bao Yi Xue Ban ; 47(5): 842-5, 2015 Oct 18.
Artigo em Chinês | MEDLINE | ID: mdl-26474628

RESUMO

OBJECTIVE: To establish a new approach to synthesis of diethyl N-[4-[(2,4-diaminopyrido[3,2-d]pyrimidin-6-yl)methylamino]benzoyl]-L-glutamate. METHODS: Target compound (5) was synthesized by the use of (2,4-dioxo-tetrahydropyridopyrimidin-6-yl)methyl acetate (1) as starting material via hydrolysis, chlorination, condensation with diethyl (p-aminobenzoyl)glutamate and aminolysis. RESULTS: A new approach to synthesis of diethyl N-[4-[(2,4-diaminopyrido[3,2-d]pyrimidin-6-yl)methylamino]benzoyl]-L-glutamate was established. This synthetic route has hydrolysis reaction, chlorination, diethyl N-(p-aminobenzoyl)-L-glutamate condensation reaction and ammonolysis reaction. The total yield is 36.7%.The structures of those compounds have identified by 1H nuclear magnetic resonance, 13C nuclear magnetic resonance and mass spectrometry. This synthetic route avoid the unstable brominated reaction product and improves the harsh condition of ammonolysis reaction. CONCLUSION: The new synthetic route has improved the reaction condition and the stability of the intermediate, and increased the extent of the derivative compounds, which has great significance to anti-folic acid of anti-tumor inhibitor synthesis.


Assuntos
Glutamatos/síntese química , Espectroscopia de Ressonância Magnética
11.
Clin Chim Acta ; 447: 66-70, 2015 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-26032866

RESUMO

BACKGROUND: Serum soluble CD40 ligand (sCD40L) concentrations are increased in patients with nasopharyngeal carcinoma (NPC). This study further evaluated the relationship between plasma sCD40L concentrations and long-term survival of NPC. METHODS: Plasma sCD40L concentrations of 312 patients and 312 healthy controls were determined using an ELISA. The associations of plasma sCD40L concentrations with 5-year overall survival, progression-free survival, distant metastasis-free survival, and locoregional relapse-free survival were investigated by univariate and multivariate analyses. RESULTS: Plasma sCD40L concentrations were substantially higher in patients than in healthy subjects and also correlated highly with tumor classification, lymph node classification and tumor node metastasis stage. sCD40L emerged as an independent predictor for 5-year overall survival, progression-free survival, distant metastasis-free survival, and locoregional relapse-free survival using univariate and multivariate Cox regression analysis. CONCLUSIONS: High plasma sCD40L concentration is correlated with stage progression of NPC as well as associated with poor survival of NPC. It is suggested that sCD40L should have the potential to be a prognostic biomarker for NPC.


Assuntos
Ligante de CD40/sangue , Ligante de CD40/química , Neoplasias Nasofaríngeas/sangue , Neoplasias Nasofaríngeas/diagnóstico , Biomarcadores Tumorais/sangue , Biomarcadores Tumorais/química , Carcinoma , Estudos de Casos e Controles , Intervalo Livre de Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Carcinoma Nasofaríngeo , Solubilidade
12.
Int J Mol Sci ; 16(2): 4136-49, 2015 Feb 16.
Artigo em Inglês | MEDLINE | ID: mdl-25690030

RESUMO

Ethylene is an important factor that stimulates Hevea brasiliensis to produce natural rubber. 1-Aminocyclopropane-1-carboxylic acid synthase (ACS) is a rate-limiting enzyme in ethylene biosynthesis. However, knowledge of the ACS gene family of H. brasiliensis is limited. In this study, nine ACS-like genes were identified in H. brasiliensis. Sequence and phylogenetic analysis results confirmed that seven isozymes (HbACS1-7) of these nine ACS-like genes were similar to ACS isozymes with ACS activity in other plants. Expression analysis results showed that seven ACS genes were differentially expressed in roots, barks, flowers, and leaves of H. brasiliensis. However, no or low ACS gene expression was detected in the latex of H. brasiliensis. Moreover, seven genes were differentially up-regulated by ethylene treatment. These results provided relevant information to help determine the functions of the ACS gene in H. brasiliensis, particularly the functions in regulating ethylene stimulation of latex production.


Assuntos
Hevea/genética , Liases/genética , Sequência de Aminoácidos , Clonagem Molecular , Etilenos/farmacologia , Genes de Plantas , Hevea/enzimologia , Liases/classificação , Liases/metabolismo , Dados de Sequência Molecular , Filogenia , Regiões Promotoras Genéticas , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Alinhamento de Sequência , Regulação para Cima/efeitos dos fármacos
13.
Clin Chim Acta ; 440: 172-6, 2015 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-25433141

RESUMO

BACKGROUND: Adrenomedullin has been identified as a tumor growth factor. However, few studies have reported its relationship with cancer survival. We evaluated the prognostic value of pretreatment plasma adrenomedullin levels in nasopharyngeal carcinoma (NPC). METHODS: Plasma adrenomedullin levels of 258 NPC patients and 100 healthy controls were determined using enzyme-linked immunosorbent assay. Adverse event was defined as tumor progression or death from any cause during 5-year follow-up. The relationships between plasma adrenomedullin levels and 5-year mortality, adverse event, tumor-free survival and overall survival were evaluated using multivariate analysis. RESULTS: Pretreatment plasma adrenomedullin levels were substantially higher in patients than in healthy subjects and were correlated highly with tumor classification, lymph node classification and tumor node metastasis stage positively. Adrenomedullin was identified as an independent predictor of 5-year mortality, adverse event, tumor-free survival and overall survival. Based on receiver operating characteristic curve analysis, pretreatment plasma adrenomedullin level had high predictive value for 5-year mortality and adverse event and had high diagnostic value for NPC. CONCLUSIONS: Adrenomedullin may be a reliable biomarker for predicting the long-term prognosis of patients with NPC and also has potential diagnostic utility for NPC.


Assuntos
Adrenomedulina/sangue , Biomarcadores Tumorais/sangue , Neoplasias Nasofaríngeas/sangue , Neoplasias Nasofaríngeas/mortalidade , Carcinoma , Estudos de Casos e Controles , Intervalo Livre de Doença , Ensaio de Imunoadsorção Enzimática , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/patologia , Neoplasias Nasofaríngeas/terapia , Valor Preditivo dos Testes , Prognóstico , Análise de Sobrevida
14.
Oncol Rep ; 28(5): 1917-23, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22941376

RESUMO

Protease-activated receptor-2 (PAR-2) has shown strong pro-angiogenesis activity physiologically and pathologically. This study aimed to explore PAR-2 regulation of pro-angiogenesis gene expression and the underlying molecular pathways in gastric cancer cells. MKN28 human gastric cancer cells were treated with trypsin, a PAR-2 activator, and subjected to real-time reverse transcription polymerase chain reaction (qRT-PCR), western blotting and ELISA for gene expression analyses. ERK1/2 phosphorylation and p38 MAP kinase inhibitors (PD98059 and SB203580, respectively) were used to block their gene activities. PAR-2 mRNA and protein were expressed in MKN-28 cells and activated by trypsin treatment. Trypsin-activated PAR-2 protein significantly enhanced expression of vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2) mRNA and protein in gastric cancer cells in a dose- and time-dependent manner. PAR-2 activation also induced the phosphorylation of ERK1/2 and p38 MAP kinase, but the ERK1/2 and p38 inhibitors blocked the activated PAR-2-induced VEGF and COX-2 expression in gastric cancer cells. PAR-2-induced expression of VEGF and COX-2 mRNA and protein in gastric cancer MKN28 cells was mediated by activation of an ERK1/2- and p38 MAP kinase-dependent pathway. Thus, PAR-2 may serve as a promising target for anti-angiogenesis therapy to treat gastric cancer.


Assuntos
Ciclo-Oxigenase 2/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Receptor PAR-2/metabolismo , Neoplasias Gástricas/metabolismo , Fatores de Crescimento do Endotélio Vascular/metabolismo , Linhagem Celular Tumoral , Ciclo-Oxigenase 2/biossíntese , Ciclo-Oxigenase 2/genética , MAP Quinases Reguladas por Sinal Extracelular/antagonistas & inibidores , MAP Quinases Reguladas por Sinal Extracelular/genética , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Flavonoides/farmacologia , Humanos , Imidazóis/farmacologia , Sistema de Sinalização das MAP Quinases , Neovascularização Patológica , Fosforilação/efeitos dos fármacos , Piridinas/farmacologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptor PAR-2/genética , Tripsina/farmacologia , Fatores de Crescimento do Endotélio Vascular/genética , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
15.
Yao Xue Xue Bao ; 47(11): 1463-9, 2012 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-23387078

RESUMO

Methionine synthase (MS, EC2.1.1.13), a key enzyme in the folate metabolism area catalyzing methyl transfer from N5-methyltetrahydrofolate to homocysteine to give tetrahydrofolate and methionine, takes a core position in folate cycle, one-carbon-unit transfer and sculpture amino acid pathways. Cobalamin-dependent methionine synthase was purified from rat liver. The enzyme was purified 609-fold to near homogeneity by batch chromatography on DE-52, anion-exchange chromatography on Q Sepharose Fast Flow and CHT-I hydroxyapatite column and was identified by SDS-PAGE and Western blotting. The enzyme activity was determined by spectrophotometric assay. In addition, the influencing factor and optimal reaction condition were performed. The steady state kinetic of rat liver methionine synthase was similar to that of other mammalian cobalamin-dependent methionine synthase which employed a Ping-Pong mechanism. The result indicated that cobalamin-dependent methionine synthase purified from rat liver is suitable for screening and studying methionine synthase specific inhibitors.


Assuntos
5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase/isolamento & purificação , 5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase/metabolismo , Fígado/química , Metotrexato/farmacologia , Quinazolinas/farmacologia , Tiofenos/farmacologia , Animais , Eletroforese em Gel de Poliacrilamida , Antagonistas do Ácido Fólico/farmacologia , Masculino , Ratos , Ratos Wistar , Tetra-Hidrofolatos/metabolismo
16.
World J Emerg Med ; 3(2): 128-34, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-25215051

RESUMO

BACKGROUND: Hypothermia is associated with poor outcome in trauma patients; however, hemorrhagic shock (HS) model with anesthetized swine was different from that of clinical reality. To identify the effects of environmental hypothermia on HS, we investigated hemodynamics and oxygen dynamics in an unanesthetized swine model of HS under simulating hypothermia environment. METHODS: Totally 16 Bama pigs were randomly divided into ambient temperature group (group A) and low temperature group (group B), 8 pigs in each group. Venous blood (30 mL/kg) was continuously withdrawn for more than 15 minutes in conscious swine to establish a hemorrhagic shock model. Pulmonary arterial temperature (Tp), heart rate (HR), mean arterial pressure (MAP), pulmonary arterial pressure (PAP), pulmonary arterial wedge pressure (PAWP), central venous pressure (CVP), cardiac output (CO), hemoglobin (Hb), saturation of mixed venous blood (SvO2) and blood gas analysis were recorded at the baseline and different hemorrhagic shock time (HST). The whole body oxygen delivery indices, DO2I and VO2I, and the O2 extraction ratio (O2ER) were calculated. RESULTS: Core body temperature in group A decreased slightly after the hemorrhagic shock model was established, and environmental hypothermia decreased in core body temperature. The mortality rate was significantly higher in group B (50%) than in group A (0%). DO2I and VO2I decreased significantly after hemorrhage. No difference was found in hemodynamics, DO2I and VO2I between group A and group B, but the difference in pH, lactic acid and O2ER was significant between the two groups. CONCLUSION: Environmental hypothermia aggravated the disorder of oxygen metabolism after hemorrhagic shock, which was associated with poor prognosis.

17.
Biosci Biotechnol Biochem ; 74(11): 2183-8, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-21071842

RESUMO

Calcium-dependent protein kinases (CDPKs), as major primary Ca(2+) sensors, have been implicated in the regulation of stress and developmental signals in plants. In this study, a novel CDPK gene, designated HbCDPK1, was isolated from Hevea brasiliensis. The HbCDPK1 cDNA had 2,400 bp with an open reading frame of 1,671 bp encoding 556 amino acids, and the deduced HbCDPK1 protein contained four characteristic domains identified in CDPKs, showing a high level of sequence similarity to CDPKs from other plants. Expression analysis revealed more significant accumulation of the transcripts of HbCDPK1 in latex than in the leaves, bark, and roots in H. brasiliensis. In addition, transcription of HbCDPK1 was strongly induced by mechanical wounding, jasmonic acid (JA), and ethephon. Recombinant HbCDPK1 was expressed in E. coli, and its activity was assayed. The assay indicated that HbCDPK1 had the kinase and Ca(2+)-binding activity in vitro as a calcium-dependent protein. The potential roles of the HbCDPK1 are discussed as to latex production and rubber biosynthesis.


Assuntos
Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Hevea/enzimologia , Compostos Organofosforados/farmacologia , Proteínas Quinases/genética , DNA Complementar/isolamento & purificação , Perfilação da Expressão Gênica , Genes de Plantas , Hevea/genética , Látex/biossíntese , Fases de Leitura Aberta/genética , Reguladores de Crescimento de Plantas , Estruturas Vegetais/química , RNA Mensageiro/análise , Borracha
18.
Biosci Biotechnol Biochem ; 73(11): 2427-31, 2009 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-19897910

RESUMO

A full-length cDNA LeCPK2 (GenBank GQ205414) from tomato (Solanum lycopersicum) encoding a calcium-dependent protein kinase (CDPK) was cloned by in silico cloning using NtCPK5 (AY971376) as a virtual probe. The deduced amino acid sequence of LeCPK2 contained the kinase, autoinhibitory, and calmodulin-like domains typical of CDPKs. Expression profiling indicated that LeCPK2 expressed predominantly in flowers and responded divergently to heat and cold stress, in which obvious mRNA accumulation was detected at 4 h under 42 degrees C stress, but no change in LeCPK2 mRNA levels was observed in 6 h at 4 degrees C. Mechanical wounding and phytohormones including ethylene, methyl jasmonate, and salicylic acid were also observed to arouse the expression of LeCPK2 in a similar pattern. mRNA accumulation was enhanced at 30 min and reached a maximum at 3 h, followed by a decrease to the normal level. All the results suggest that LeCPK2 is a novel versatile isoform of tomato CDPKs.


Assuntos
Regulação da Expressão Gênica de Plantas , Temperatura Alta , Lycopersicon esculentum/efeitos dos fármacos , Lycopersicon esculentum/genética , Reguladores de Crescimento de Plantas/farmacologia , Proteínas Quinases/genética , Sequência de Aminoácidos , Clonagem Molecular , Biologia Computacional , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Lycopersicon esculentum/citologia , Lycopersicon esculentum/fisiologia , Dados de Sequência Molecular , Especificidade de Órgãos , Proteínas Quinases/química , Estresse Mecânico , Estresse Fisiológico/genética
19.
Plant Physiol Biochem ; 47(10): 954-9, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19577934

RESUMO

Hsp70s have been shown to play important roles in helping cells to cope with adverse environments, especially in response to temperature. In this study a novel ethephon-induced Hsp gene, designated as HbHsp70, was isolated from Hevea brasiliensis. The HbHsp70 cDNA contained a 1965 bp open reading frame encoding 655 amino acids. The deduced HbHsp70 protein showed high identities to Hsp70s from other plants. Expression studies revealed more significant accumulation of HbHsp70 transcripts in leaves and stems than in roots, barks and latex. The transcription of HbHsp70 was induced by ethephon, heat treatment and low temperature stress, whereas jasmonic acid had little effects. Recombinant HbHsp70 was expressed in Escherichia coli and purified by Ni-NTA affinity chromatography. Measuring the light scattering of luciferase (Luc) revealed that HbHsp70 prevents the aggregation of luc during high-temperature stress. In vitro experiments showed that HbHsp70 had protective functions not only against heat stress but also against chilling stress. All these data suggest that HbHsp70 may play roles in responses to heat shock and low temperature in H. brasiliensis.


Assuntos
Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Proteínas de Choque Térmico HSP70/genética , Hevea/genética , Compostos Organofosforados/farmacologia , Proteínas de Plantas/genética , Sequência de Aminoácidos , Clonagem Molecular , Temperatura Baixa , DNA Complementar/química , DNA Complementar/genética , Eletroforese em Gel de Poliacrilamida , Perfilação da Expressão Gênica , Proteínas de Choque Térmico HSP70/metabolismo , Hevea/metabolismo , Temperatura Alta , Chaperonas Moleculares/metabolismo , Dados de Sequência Molecular , Reguladores de Crescimento de Plantas/farmacologia , Proteínas de Plantas/metabolismo , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos
20.
Beijing Da Xue Xue Bao Yi Xue Ban ; 40(4): 443-5, 2008 Aug 18.
Artigo em Chinês | MEDLINE | ID: mdl-18677396

RESUMO

OBJECTIVE: To find a best synthesis method of diethyl N-[p-(methylamino)benzoyl]-L-glutamate(1). METHODS: Using diethyl N-(p- aminobenzoyl)-L-glutamate (2) as a starting material via a one-pot process of benzylation and methylation, the diethyl N-[p-(methylamino)benzoyl ]-L-glutamate (3) is prepared in high yield. Then compound 1 was conveniently obtained from debenzylation of compound 3 by Pd/C. RESULTS: The target compound was obtained through 2-step reaction in 88% overall yield in only 6 h, and its structure was identified by 1HNMR and MS. CONCLUSION: It is a convenient and efficient method of preparation of diethyl N-[p-(methylamino)benzoyl]-L-glutamate.


Assuntos
Glutamatos/química , Glutamatos/síntese química
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