RESUMO
PURPOSE: Aspirin could reduce the risk of cancer metastasis. Circulating tumor cells (CTCs) are a key factor of cancer metastasis, but no evidence has revealed how aspirin affects CTCs and its epithelial-mesenchymal transition (EMT). Here, we conducted a clinical trial to investigate how aspirin affects CTCs in metastatic colorectal cancer (MCC) and breast cancer patients (MBC). METHODS: The trial is retrospective registered at clinicaltrials.gov (NCT02602938). The eligible patients are given 100 mg aspirin q.d. for 8 weeks, and CTCs are evaluated at baseline, 4 and 8 weeks for absolute number, phenotype (epithelial type, E+, mesenchymal type, M+, and biophenotypic type, B+), and vimentin expression. RESULTS: Data on 21 MCC and 19 MBC patients are analyzed, and it revealed that the CTC numbers decreased with aspirin treatment in MCC (p < 0.001) but not MBC (p = 0.0532); besides, ratio of E+ CTCs increased (p = 0.037) and M+ CTCs decreased at 2 months in MCC (p = 0.013), but neither the ratio of E+ or M+ CTCs changes significantly in MBC; vimentin expression of M+ CTCs is higher than E+ and B+ CTCs either in MBC or MCC patients at baseline (p < 0.01); and aspirin suppresses the vimentin expression in M+ (p = 0.002)and B+ (p = 0.006) CTCs of MCC and M+ CTCs of MBC (p = 0.004); besides it find vimentin expression in B+ (p = 0.004) or M+ (p < 0.001), CTCs are markedly decreased in patients with total CTC numbers declined. CONCLUSION: Aspirin could decrease CTCs numbers and block EMT transition in MCC patients and part of MBC patients.
Assuntos
Aspirina/administração & dosagem , Neoplasias da Mama/patologia , Neoplasias Colorretais/patologia , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Células Neoplásicas Circulantes/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anti-Inflamatórios não Esteroides/administração & dosagem , Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/sangue , Neoplasias da Mama/tratamento farmacológico , Neoplasias Colorretais/sangue , Neoplasias Colorretais/tratamento farmacológico , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica , Células Neoplásicas Circulantes/efeitos dos fármacos , Prognóstico , Estudos Prospectivos , Estudos Retrospectivos , Vimentina/metabolismo , Adulto JovemRESUMO
This study aimed to explore the protective effect of quercetin on acute lung injury (ALI) in rats with sepsis and the related mechanism. Rats were administered different doses of quercetin intraperitoneally, and blood samples and lung tissue were collected at 24 h after treatment. Arterial blood gases, lung water content, protein content, and cell counts in bronchoalveolar lavage fluid (BALF) were measured. Morphological changes in lung tissue pathology were observed under a light microscope. Serum intercellular adhesion molecule (ICAM)-1 and macrophage inflammatory protein 2 (MIP-2) levels were detected and ICAM-1 and MIP-2 mRNA expression in lung tissue was determined. Compared with that in the control model group, arterial blood gases, lung water content, protein content, and cell counts in BALF improved in the high- and low-dose quercetin groups (P < 0.05), with maximal improvement observed for the high-dose quercetin (P < 0.05). Lesions on the lungs improved in the high- and low-dose quercetin groups than those in the control model group, and the high-dose quercetin group showed better improvement than the low-dose group (P < 0.05). Compared with that in the sham-operated group, both serum and lung tissue ICAM-1 and MIP-2 expression increased significantly in the model group (P < 0.05). The quercetin groups presented lower ICAM-1 and MIP-2 expression than the control model group, with the lowest expression observed in the high-dose group (P < 0.05). Quercetin may protect against ALI in rats with sepsis by inhibiting ICAM-1 and MIP-2 expression.
Assuntos
Lesão Pulmonar Aguda/tratamento farmacológico , Quimiocina CXCL2/metabolismo , Molécula 1 de Adesão Intercelular/metabolismo , Substâncias Protetoras/farmacologia , Quercetina/farmacologia , Sepse/tratamento farmacológico , Lesão Pulmonar Aguda/metabolismo , Lesão Pulmonar Aguda/microbiologia , Animais , Líquido da Lavagem Broncoalveolar , Avaliação Pré-Clínica de Medicamentos , Pulmão/metabolismo , Pulmão/patologia , Masculino , Substâncias Protetoras/uso terapêutico , Quercetina/uso terapêutico , Ratos Wistar , Sepse/metabolismoRESUMO
This study aimed to investigate cytosine methylation profiles in different tobacco (Nicotiana tabacum) cultivars grown in China. Methylation-sensitive amplified polymorphism was used to analyze genome-wide global methylation profiles in four tobacco cultivars (Yunyan 85, NC89, K326, and Yunyan 87). Amplicons with methylated C motifs were cloned by reamplified polymerase chain reaction, sequenced, and analyzed. The results show that geographical location had a greater effect on methylation patterns in the tobacco genome than did sampling time. Analysis of the CG dinucleotide distribution in methylation-sensitive polymorphic restriction fragments suggested that a CpG dinucleotide cluster-enriched area is a possible site of cytosine methylation in the tobacco genome. The sequence alignments of the Nia1 gene (that encodes nitrate reductase) in Yunyan 87 in different regions indicate that a C-T transition might be responsible for the tobacco phenotype. T-C nucleotide replacement might also be responsible for the tobacco phenotype and may be influenced by geographical location.
Assuntos
Citosina/metabolismo , Metilação de DNA/genética , Nicotiana/genética , Nicotiana/metabolismo , Polimorfismo Genético/genética , China , Ilhas de CpG/genética , DNA de Plantas/genética , Genes de Plantas/genética , Genoma de Planta/genética , Nitrato Redutase/genética , Reação em Cadeia da Polimerase/métodosRESUMO
The aim of this study was to analyze the effect of linker length on the expression and biological activity of recombinant protein onconase (ONC) in fusion with human serum albumin (HSA) in Pichia pastoris. Four flexible linkers with different lengths namely Linker L0, L1: (GGGGS)1, L2: (GGGGS)2, and L3:(GGGGS)3 were inserted into the fusion gene and referred to as HSA-n-ONC, where N = 0, 5, 10, or 15. The sequence of the fusion gene HSA-ONC was designed based on the GC content and codon bias in P. pastoris; the signal peptide of albumin was used as the secretion signal. Gene sequences coding for the fusion protein with different linkers were inserted into pPICZα-A to form recombinant plasmids pPICZα-A/HSA-n-ONC, which were then transformed into P. pastoris X-33 for protein expression. Ideal conditions for expression of the fusion proteins were optimized at a small scale, using shake flasks before proceeding to mass production in 10-L fermenters. The recombinant fusion proteins were purified by aqueous two-phase extraction coupled with DEAE anion exchange chromatography, and their cytotoxic effect on the tumor cell was evaluated by the sulforhodamine B assay. The results showed that the expressed amount of fusion proteins had no significant relationship with the length of different linkers and rHSA-0-ONC had no cytotoxic effect on the tumor cells. While rHSA-5-ONC and rHSA-10-ONC had a weak cytotoxic effect, rHSA-15-ONC could kill various tumor cells in vitro. In summary, the biological activity of the fusion protein gradually improved with increasing length of the linker.
Assuntos
Proteínas de Anfíbios/genética , Antineoplásicos/farmacologia , Clonagem Molecular/métodos , Pichia/genética , Proteínas Recombinantes de Fusão/genética , Ribonucleases/genética , Proteínas de Anfíbios/biossíntese , Proteínas de Anfíbios/farmacologia , Animais , Antineoplásicos/química , Antineoplásicos/metabolismo , Técnicas de Cultura Celular por Lotes , Reatores Biológicos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Expressão Gênica , Humanos , Concentração Inibidora 50 , Extração Líquido-Líquido , Pichia/metabolismo , Plasmídeos/química , Plasmídeos/metabolismo , Engenharia de Proteínas , Sinais Direcionadores de Proteínas , Rana pipiens/metabolismo , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/farmacologia , Rodaminas/química , Ribonucleases/biossíntese , Ribonucleases/farmacologia , Albumina Sérica/biossíntese , Albumina Sérica/genética , Relação Estrutura-Atividade , Transformação GenéticaRESUMO
Clostridium difficile is the most common cause of hospital-acquired diarrhea in patients treated with antibiotics, chemotherapeutic agents, and other drugs that alter the normal equilibrium of the intestinal flora. A better understanding of the risk factors for C. difficile-associated disease (CDAD) could be used to reduce the incidence of CDAD and the costs associated with its treatment. The aim of this study was to identify the risk factors for CDAD in a cohort of Chinese patients in a Beijing hospital. Medical charts of a total of 130 inpatients (62 males and 68 females) with hospital-acquired diarrhea (45 with CDAD; 85 without CDAD) were retrospectively reviewed. C. difficile toxins A and B were detected in fecal samples using enzyme-linked fluorescence assays. The drugs used by patients with and without CDAD before the onset of diarrhea were compared. Factors that differed significantly between the two groups by univariate analysis were analyzed by multivariate analysis using a logistic regression model. Multivariate analysis showed that cephalosporin treatment was associated with a significantly higher risk of CDAD in hospitalized patients, while treatment with glycopeptides was significantly associated with a reduction in CDAD (P<0.001 for cephalosporin; P=0.013 for glycopeptides). Our data confirmed previous findings that empirical treatment with cephalosporins is positively associated with CDAD compared to individuals using other CDAD-related drugs. Additionally, we showed that treatment with glycopeptides was negatively associated with CDAD, compared to individuals using other CDAD-related drugs.
Assuntos
Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Antibacterianos/efeitos adversos , Clostridioides difficile/patogenicidade , Infecção Hospitalar/microbiologia , Diarreia/microbiologia , Enterocolite Pseudomembranosa/microbiologia , Proteínas de Bactérias/isolamento & purificação , Toxinas Bacterianas/isolamento & purificação , Cefalosporinas/efeitos adversos , China/epidemiologia , Infecção Hospitalar/epidemiologia , Enterocolite Pseudomembranosa/epidemiologia , Enterotoxinas/isolamento & purificação , Fezes/microbiologia , Glicopeptídeos/uso terapêutico , Incidência , Modelos Logísticos , Análise Multivariada , Estudos Retrospectivos , Fatores de Risco , Estatísticas não ParamétricasRESUMO
Clostridium difficile is the most common cause of hospital-acquired diarrhea in patients treated with antibiotics, chemotherapeutic agents, and other drugs that alter the normal equilibrium of the intestinal flora. A better understanding of the risk factors for C. difficile-associated disease (CDAD) could be used to reduce the incidence of CDAD and the costs associated with its treatment. The aim of this study was to identify the risk factors for CDAD in a cohort of Chinese patients in a Beijing hospital. Medical charts of a total of 130 inpatients (62 males and 68 females) with hospital-acquired diarrhea (45 with CDAD; 85 without CDAD) were retrospectively reviewed. C. difficile toxins A and B were detected in fecal samples using enzyme-linked fluorescence assays. The drugs used by patients with and without CDAD before the onset of diarrhea were compared. Factors that differed significantly between the two groups by univariate analysis were analyzed by multivariate analysis using a logistic regression model. Multivariate analysis showed that cephalosporin treatment was associated with a significantly higher risk of CDAD in hospitalized patients, while treatment with glycopeptides was significantly associated with a reduction in CDAD (P<0.001 for cephalosporin; P=0.013 for glycopeptides). Our data confirmed previous findings that empirical treatment with cephalosporins is positively associated with CDAD compared to individuals using other CDAD-related drugs. Additionally, we showed that treatment with glycopeptides was negatively associated with CDAD, compared to individuals using other CDAD-related drugs.
Assuntos
Antibacterianos/efeitos adversos , Clostridioides difficile/patogenicidade , Infecção Hospitalar/microbiologia , Diarreia/microbiologia , Enterocolite Pseudomembranosa/microbiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteínas de Bactérias/isolamento & purificação , Toxinas Bacterianas/isolamento & purificação , Cefalosporinas/efeitos adversos , China/epidemiologia , Infecção Hospitalar/epidemiologia , Enterocolite Pseudomembranosa/epidemiologia , Enterotoxinas/isolamento & purificação , Fezes/microbiologia , Feminino , Glicopeptídeos/uso terapêutico , Humanos , Incidência , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Estudos Retrospectivos , Fatores de Risco , Estatísticas não ParamétricasRESUMO
Fifteen new polymorphic microsatellite loci were developed for the cuttlefish Sepiella maindroni. In 32 individuals from a wild population of coastal Ningde, Fujian Province, China, the number of alleles at these loci varied between 2 and 12, with an average of 5.86. The mean observed and expected heterozygosities were 0.6917 and 0.5993, respectively. Among these polymorphic microsatellite loci, 4 (SM2, SM19, SM40, and SM81) significantly deviated from Hardy-Weinberg equilibrium after sequential Bonferroni's correction. All of them were in linkage equilibrium. These microsatellite loci would be useful for evaluating the effect of releasing on extant S. maindroni populations as well as for investigating genetic diversity and population structure of this species.
Assuntos
Decapodiformes/genética , Repetições de Microssatélites , Polimorfismo Genético , Animais , Ligação Genética , Marcadores Genéticos , Heterozigoto , População/genéticaRESUMO
Our previous studies have shown that tandem Alu repeats inhibit green fluorescent protein (GFP) gene expression when inserted downstream of the GFP gene in the pEGFP-C1 vector. We found that the 22R sequence (5'-GTGAAAAAAATGCTTTATTTGT-3') from the antisense PolyA (240 bp polyadenylation signal) of simian virus 40, eliminated repression of GFP gene expression when inserted between the GFP gene and the Alu repeats. The 22R sequence contains an imperfect palindrome; based on RNA structure software prediction, it forms an unstable stem-loop structure, including a loop, a first stem, a bulge, and a second stem. Analysis of mutations of the loop length of the 22R sequence showed that the three-nucleotide loop (wild-type, 22R) induced much stronger GFP expression than did other loop lengths. Two mutations, 4TMI (A7âT, A17âT) and 5AMI (A6âT, T18âA), which caused the base type changes in the bulge and in the second stem in the 22R sequence, induced stronger GFP gene expression than 22R itself. Mutation of the bulge base (A17âT), leading to complete complementation of the stem, caused weaker GFP gene expression. Sequences without a palindrome (7pieA, 5'-GTGAAAAAAATG CAAAAAAAGT-3', 7pieT, 5'-GTGTTTTTTTTGCTTTTTTTGT-3') did not activate GFP gene expression. We conclude that an imperfect palindrome affects and can increase GFP gene expression.
Assuntos
Regulação Viral da Expressão Gênica , Proteínas de Fluorescência Verde/genética , Sinais de Poliadenilação na Ponta 3' do RNA , Vírus 40 dos Símios/genética , Proteínas Virais/genética , Elementos Alu/genética , Sequência de Bases , Linhagem Celular Tumoral , Primers do DNA , Genes Reporter , Vetores Genéticos , Proteínas de Fluorescência Verde/biossíntese , Células HeLa , Humanos , Sequências Repetidas Invertidas , Plasmídeos , Poli A/genética , RNA Viral/genética , RNA Viral/metabolismo , Análise de Sequência de DNA , Vírus 40 dos Símios/metabolismoRESUMO
Thyroid hormone receptors (TR) are members of the nuclear receptor superfamily. There are at least two TR isoforms, TRα and TRß. The TRα isoform plays a critical role in mediating the action of thyroid hormone in adipose tissue. We mapped the porcine TRα gene to chromosome 12 p11-p13, by using the ImpRH panel. We examined tissue-localization of TRα and determined expression patterns of TRα in porcine adipose tissue with quantitative real-time PCR. TRα was expressed in all tissues, including heart, liver, spleen, stomach, pancreas, brain, small intestine, skeletal muscle, and subcutaneous adipose tissue. In the adipose tissue, the expression of TRα decreased postnatally. Compared to Yorkshire pigs, Jinhua pigs had significantly lower expression levels of TRα gene in the subcutaneous fat tissue. The expression levels of ß2-AR, HSL and ATGL were also significantly lower in Jinhua pigs than in Yorkshire pigs. However, no significant differences in PPARγ and SREBP-1C expression levels were found between Jinhua and Yorkshire pigs. Incubation of porcine adipose tissue explants with high doses of isoproterenol (100 and 1000 nM) significantly increased the expression levels of TRα. We conclude that there is considerable evidence that TRα plays an important role in fat deposition in porcine adipose tissue.