RESUMO
The aim of the present study is to explore the effect of IL-6 gene polymorphisms on the development of keloid scar (KS) in the Chinese Han population. Genotyping of IL-6 was performed by the polymerase chain reaction (PCR), followed by restriction fragment length polymorphism assays (PCR-RFLP). Serum level of IL-6 was measured using enzyme-linked immunosorbent assay (ELISA). Results indicated that when the IL-6 -572 CC homozygote genotype was used as the reference group, the GG genotype was found to be associated with a significantly increased risk of KS (GG vs CC: OR = 2.097, 95%CI â=â1.100-3.995, P â= â0.025). When the IL-6 -572 C allele was used as the reference group, the G allele was found to be associated with significantly increased risk of KS (G vs C: OR = â1.317, 95%CI â=â1.002-1.730, Pâ=â0.048). Furthermore, we observed a marked increase in serum IL-6 levels in KS patients with GG genotypes when compared to KS patients harboring the CC genotype. In conclusion, our results suggest that IL-6 gene polymorphism was associated with keloid scars in the southeastern Chinese Han population.
Assuntos
Interleucina-6/genética , Queloide/genética , Polimorfismo de Nucleotídeo Único , Adolescente , Adulto , Alelos , Estudos de Casos e Controles , China , Feminino , Genótipo , Humanos , Interleucina-6/sangue , Queloide/sangue , Masculino , Pessoa de Meia-IdadeRESUMO
We investigated the extraction of Toona sinensis fruit proteins and preliminarily characterized their physicochemical properties. The results showed that optimal extraction occurred under conditions of pH 10.5, a duration of 40 min, a liquid-to-solid ratio of 25:1, and a temperature of 40°C by an orthogonal design using T. sinensis fruit protein as the index and single factor. The total nitrogen content was 13.8 g/100 g and included 17 different amino acids. The glutamate level was highest at 35.37%, followed by arginine at 15.31%. The isoelectric point of T. sinensis fruit protein was between 6.8 and 10.0 with a typical absorption peak by infrared chromatography. Three protein bands were analyzed using SDS-polyacrylamide gel electrophoresis, with relative molecular weights of 55, 51, and 22 kDa. This study provides a theoretical basis for the comprehensive utilization of T. sinensis fruit by further investigating the biological activity of its proteins.
Assuntos
Frutas/química , Meliaceae/química , Extratos Vegetais/química , Proteínas de Plantas/química , Proteômica/métodosRESUMO
Axillary branching is controlled by a very complex mechanism involving various endogenous and environmental factors. Previous studies have shown that Tb1/BRC1 is the point of integration in the network of molecular mechanisms regulating axillary branching in plants. In this study, we cloned the Tb1/BRC1 ortholog, NtBRC1, from Nicotiana tabacum and functionally analyzed its role in the control of axillary branching in tobacco. Overexpression of NtBRC1 resulted in significant retardation of axillary branching, and downregulation of this gene resulted in significant acceleration of axillary branching after decapitation. This indicates a negative role for this gene in the regulation of axillary branching. In-line with previous reports, NtBRC1 was found to be expressed predominantly in axillary buds. Additionally, as expected, expression was decreased 8 h following decapitation, which further confirms its role in the suppression of axillary branching. Furthermore, the expression of NtBRC1 was significantly downregulated by cytokinin, but was not affected by GR24, a synthetic strigolactone. Based on the data collected in the present study, we demonstrate that NtBRC1 negatively regulates axillary branching in tobacco after decapitation and functions downstream of the cytokinin signaling pathway inside axillary buds.
Assuntos
Nicotiana/fisiologia , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Clonagem Molecular , Citocininas/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Lactonas/farmacologia , Nicotiana/genéticaRESUMO
Rectal cancer is a commonly observed tumor in clinics, and epithelial-mesenchymal transition (EMT) is very important for tumor invasion and metastasis. We established a rectal cancer HCT-116 cell hypoxia model and detected cell proliferation, invasion, and EMT-related protein expression in this model, aiming to analyze the effect of hypoxia on rectal cancer cell EMT. Rectal cancer cell line HCT-116 was cultured in normoxic, hypoxic, or anaerobic environment, and hypoxia-inducible factor-1α (HIF-1α) mRNA expression was detected in the cells by real-time PCR. Cell proliferation was tested by MTT assay; cell invasion was determined by transwell assay, and HIF-1α, epithelial-cadherin, and Snail protein levels were evaluated by western blot analysis. HIF-1α mRNA level significantly increased in the anaerobic group compared to that in the normoxic and hypoxic groups (P < 0.05). HCT-116 cell proliferation in the anaerobic group was obviously higher than that in the other two groups, with the hypoxic group showing stronger proliferative ability than the normoxic group (P < 0.05). Compared to the normoxic group, the HCT-116 cells demonstrated enhanced cell invasion and migration in hypoxic and anaerobic groups. HIF-1α and Snail expressions were upregulated, whereas epithelial-cadherin expression had declined in the hypoxic and anaerobic groups, compared to those in the normal control (P < 0.05). Therefore, hypoxia promoted rectal cancer cell progress by increasing HIF-1α to induce EMT.
Assuntos
Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Neoplasias Retais/patologia , Fatores de Transcrição da Família Snail/metabolismo , Hipóxia Celular , Movimento Celular , Proliferação de Células , Transição Epitelial-Mesenquimal , Regulação Neoplásica da Expressão Gênica , Células HCT116 , Humanos , Neoplasias Retais/genética , Neoplasias Retais/metabolismoRESUMO
MicroRNAs (miRNAs) are key regulators of gene expression and play an important role in the development and progression of various diseases including esophageal squamous cell carcinoma (ESCC). In this study, we determined whether a polymorphism at the miR-214 binding site in the 3'-untranslated region (3'-UTR) of the methylenetetrahydrofolate reductase gene (MTHFR) is associated with susceptibility to ESCC. A total of 448 ESCC cases and 460 gender- and age-matched subjects were recruited for the study. The genotypes of the rs114673809 single nucleotide polymorphism (SNP) were determined by polymerase chain reaction sequencing. Associations between genotypes of MTHFR rs114673809 and ESCC risk were determined using logistic regression analyses. In the recessive model, when the MTHFR rs114673809 GG homozygote genotype was used as the reference group, the GA genotype was not associated with the risk of ESCC (GA vs GG: OR = 1.261, 95%CI = 0.960-1.657, P = 0.110), but the AA genotype was associated with increased risk of ECSS (AA vs GG: OR = 1.752, 95%CI = 1.076-2.853, P = 0.027). Additionally, the rs114673809 A allele carriers also showed a 1.286-fold increased ESCC risk compared with those carrying the rs114673809 G allele genotype. Furthermore, we observed a significant increase in plasma homocysteine levels in ESCC cases carrying the AA genotype relative to ESCC cases carrying the GG genotype. Our data demonstrate that a polymorphism at the miR-214 binding site in the 3'-UTR of MTHFR is an ESCC susceptibility SNP in the Chinese population.
Assuntos
Carcinoma de Células Escamosas/genética , Neoplasias Esofágicas/genética , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , MicroRNAs/genética , Regiões 3' não Traduzidas , Adulto , Idoso , Alelos , Povo Asiático/genética , Sítios de Ligação/genética , Carcinoma de Células Escamosas/enzimologia , Carcinoma de Células Escamosas/metabolismo , Estudos de Casos e Controles , Neoplasias Esofágicas/enzimologia , Neoplasias Esofágicas/metabolismo , Carcinoma de Células Escamosas do Esôfago , Feminino , Predisposição Genética para Doença , Genótipo , Humanos , Masculino , Metilenotetra-Hidrofolato Redutase (NADPH2)/metabolismo , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo ÚnicoRESUMO
The objective of this study was to observe the protective effect of the n-butyl alcohol phase of Toona sinensis seed extract on the kidneys of diabetic nephropathy (DN) rats and its preliminary mechanism. Male wistar rats were administered a normal or high-fat diet for 1 month. DN rats were divided into a model group and a petroleum ether phase of T. sinensis seed extract intervention group. The intervention group was administered 5 mg·100 g-1·day-1 extract. After treatment for 10 weeks, the rats were sacrificed and blood samples and the renal cortex were collected. Biochemical indicators in the serum and renal indices were assessed. Pathological changes of the renal tissues were also determined. Changes in the renal structure and protein levels were detected. Compared with the normal group, the blood glucose, urinary albumin, renal index, and oxidative stress index were sharply increased in the model group. The protein levels of TGF-b1, collagen IV, and connective tissue growth factor (CTGF) were increased. Compared with the model group, the n-butyl alcohol phase of T. sinensis seed extract significantly reduced the blood glucose, urinary albumin, renal index, oxidative stress index, serum creatinine, and urea nitrogen levels. The renal pathology abnormality was improved in DN rats. The protein levels of TGF-b1, collagen IV, and CTGF were increased. The expression of TGF-b1, collagen IV, and CTGF decreased. In conclusion, the n-butyl alcohol phase of T. sinensis seed extract has protective effects on DN rats via the inhibition of oxidative stress and protein expression of TGF-b1, collagen IV, and CTGF.
Assuntos
1-Butanol/química , Nefropatias Diabéticas/tratamento farmacológico , Nefropatias Diabéticas/patologia , Rim/patologia , Meliaceae/química , Extratos Vegetais/química , Extratos Vegetais/uso terapêutico , Sementes/química , Animais , Rim/efeitos dos fármacos , Masculino , Ratos , Ratos WistarRESUMO
We investigated the possible association between two single nucleotide polymorphisms of IL10 (-1082A/G and -592C/A) and susceptibility to ischemic stroke. In total, 335 patients with proven ischemic stroke and 335 control subjects were recruited from Xinxiang Central Hospital between March 2013 and May 2015. The IL10 -1082A/G and -529C/A polymorphisms were investigated by polymerase chain reaction-restriction fragment length polymorphism. When compared with the control subjects, patients with ischemic stroke were more likely to be male, have a habit of tobacco smoking, have higher BMI, have hypertension or diabetes mellitus, and have higher levels of TC, LDL-C, HDL-C, and TG. The multivariate logistic regression analyses revealed that the AA genotype of IL10 -1082A/G was significantly associated with development of ischemic stroke in a Chinese population compared with the GG genotype (OR = 1.93, 95%CI = 1.15-3.25). In the dominant model, the association between GA+AA genotype of IL10 -1082A/G and risk of ischemic stroke was also significant compared with the GG genotype, and the adjusted OR (95%CI) for the GA+AA genotype was 1.41 (1.02-1.94). Thus, our study suggests that IL10 gene polymorphisms contribute to the development of ischemic stroke.
Assuntos
Isquemia Encefálica/genética , Interleucina-10/genética , Polimorfismo de Nucleotídeo Único , Acidente Vascular Cerebral/genética , Isquemia Encefálica/epidemiologia , Estudos de Casos e Controles , China , Diabetes Mellitus/epidemiologia , Feminino , Humanos , Hipertensão/epidemiologia , Masculino , Pessoa de Meia-Idade , Fatores Sexuais , Fumar/epidemiologia , Acidente Vascular Cerebral/epidemiologiaRESUMO
A serine/threonine protein kinase gene (NrSTK) was cloned from Nicotiana repanda based on the sequence of a previously isolated resistance gene analog (RGA). Expression of RGA was induced by challenge with the pathogen black shank. The NrSTK gene was predicted to encode a protein kinase that contained an ATP binding site at residues 41-69 and a serine/threonine protein kinase activation sequence spanning the region 161-173. Overexpression of NrSTK in the susceptible tobacco variety Honghuadajinyuan significantly enhanced resistance to black shank, indicating that NrSTK plays a role in incompatibility reactions between tobacco and the pathogen. Characterization of NrSTK will help elucidate the molecular mechanisms involved in black shank resistance in N. repanda.
Assuntos
Resistência à Doença/genética , Nicotiana/genética , Doenças das Plantas/genética , Proteínas Serina-Treonina Quinases/genética , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , DNA Complementar , Dados de Sequência Molecular , Filogenia , Plantas Geneticamente Modificadas , Alinhamento de Sequência , Análise de Sequência de DNARESUMO
Testis-specific serine kinases (TSSKs) are a family of serine/threonine kinases highly expressed in the testes that are responsible for regulating many spermatogenesis-related protein activities. Mutations in this family have a positive relationship with oligospermia and azoospermia in human and mouse. Here, five members of the TSSK family from a Banna mini-pig inbred line (BMI) were cloned, sequenced, and characterized. The full-length coding sequences of BMI TSSKs varied from 807 (TSSK3) to 1095 bp (TSSK1) and encoded 268 to 364 amino acids with molecular weights in the range 30.11 to 41.34 kDa. Following comparison with TSSK4 genes in other species, BMI TSSK4 was found to contain three alternatively spliced variants, inform1, inform 3, and inform 4. BMI TSSK1 and TSSK2 are co-localized on the Sus scrofa chromosome (SSC) 14, and consist of a single exon; TSSK3, TSSK4, and TSSK6 are on SSC6, SSC7, and SSC2, and consist of two, four, and one exon, respectively. Multiple protein sequence alignment and phylogenetic analysis showed that the regions spanning the S_TKc domains were more conserved between pig and other animals: with TSSK1 and TSSK2 and TSSK3 and TSSK6 displaying the greatest degree of homology across species, and the TSSK4 protein clearly distinct from other members. Multi-tissue RT-PCR showed BMI TSSK1, TSSK3, and TSSK4 were only expressed in the testes and seminal vesicle, TSSK2 was confined to testes only, while TSSK6 was expressed widely in adult tissues but was highest in the testes.
Assuntos
Família Multigênica/genética , Proteínas Serina-Treonina Quinases/genética , Espermatogênese/genética , Testículo/crescimento & desenvolvimento , Adulto , Sequência de Aminoácidos/genética , Animais , Clonagem Molecular , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Masculino , Camundongos , Especificidade de Órgãos/genética , Filogenia , Proteínas Serina-Treonina Quinases/biossíntese , Glândulas Seminais/crescimento & desenvolvimento , Glândulas Seminais/metabolismo , Suínos , Porco Miniatura , Testículo/metabolismoRESUMO
The lymphotoxin beta receptor (LTßR) is a member of the tumor necrosis factor family of receptors (TNFR). It plays a role in regulating lymphoid organogenesis and homeostasis of the immune system. In the present study, the full coding region of a putative LTßR gene of Sus scrofa was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and cloned for the first time (accession Nos. JX457347 and AFU74012). In addition, analysis of the tissue expression profile was carried out via RT-PCR. The full-length coding region of porcine LTßR had 1266 nucleotides (molecular weight, 45.61 kDa; pI, 5.71) and encoded 421 amino acids. Bioinformatic prediction indicates that LTßR belongs to the TNFR superfamily and contains a TNFR domain. The sequence homology analysis revealed that the amino acid sequences of S. scrofa LTßR had 82.9, 82.4, 81.3, 80.5, 78.7, 74.6, and 73.0% identity with those of Equus caballus, Canis lupus, Ailuropoda melanoleuca, Oryctolagus cuniculus, Bos taurus, Mus musculus, and Homo sapiens, respectively. The phylogenetic tree based on the amino acid sequences of LTßR from 8 species revealed that S. scrofa was more closely related to E. caballus, C. lupus, and A. melanoleuca. RT-PCR analysis showed that the porcine LTßR gene was differentially expressed (e.g., high, moderate, low, or nonexistent) in various tissues (e.g., prostate, pituitary, brainstem, and esophagus, respectively). This may be related to differences in the regulation of LTßR in the different tissues.