RESUMO
Coilia ectenes is a commercially important fishery species in China. C. ectenes taihuensis is an endemic and dominant species found in Taihu Lake of China. When compared with C. ectenes, C. ectenes taihuensis lacks anadromous behavior, and can independently grow and reproduce in Taihu Lake. In this study, the mitochondrial DNA control region (D-loop) sequences were employed to investigate the genetic diversity and population structure of C. ectenes taihuensis. Sixty-eight individuals collected from 4 localities in Taihu Lake were examined. Results indicated that in the 887-bp D-loop region, seventy-seven (8.68%) sites were variant, contributing to 53 distinct haplotypes. Although the population haplotype diversity (Hd = 0.971 to 1.000) was generally high, the nucleotide diversity (π = 0.616 to 0.731%) was relatively low among the 4 populations. Additionally, the genetic distances ranged from 0.62 to 0.74% within the populations and from 0.67 to 0.74% between the populations. The neighbor-joining tree indicated that a distinct distribution of phylogenetic structure existed among haplotypes. Analysis of molecular variance and FST statistics suggested that a divergence existed among populations in 4 localities, indicating that gene communication might have occurred among those populations. Furthermore, neutral tests and analysis of mismatch distribution reflected that C. ectenes taihuensis might have undergone a population expansion during the evolution process. Our study showed the population genetic diversity and structure of C. ectenes taihuensis. Results from this study might be helpful in the development and protection of fishery resource within the localities in Taihu Lake in future.
Assuntos
DNA Mitocondrial/genética , Peixes/genética , Polimorfismo Genético , Animais , China , Evolução Molecular , Genética Populacional , Haplótipos , Lagos , FilogeniaRESUMO
A combination of phenotypic characterization and molecular markers may provide reliable information on new plant varieties and elucidate the conservation status of rare species. Five newly developed Magnolia wufengensis cultivars, an endangered plant species endemic to Hubei Province, China, possess more distinctive phenotypes than common Magnolia cultivars. With reference to a wild species population of M. wufengensis and a population of Magnolia denudata, morphological traits of flower organs, simple sequence repeat (SSR), and sequence-related amplified polymorphism (SRAP) markers were used. In the morphological study, six traits of floral organs were investigated and their relationships were analyzed between cultivars. In the genetic study, 9 SSR primer pairs and 10 SRAP primer combinations were screened. The five cultivars maintained a high level of genetic diversity. Genetic diversity of each M. wufengensis cultivar was much lower than that of the wild population, but was slightly higher than that of the M. denudata population. Analysis of molecular variance (AMOVA) revealed that genetic variation among populations was 20% (SRAP) and 30% (SSR), which showed a high degree of genetic differentiation among populations of the five cultivars. The dendrograms illustrated a clear separation between M. wufengensis populations and outer species, and identified two major groups among cultivars. Correlation analysis indicated a good fit between the two marker systems, but a relatively low fit between morphological and genetic traits (SRAP: r = 0.60, SSR: r = 0.52). These findings provide reliable references for the application of these molecular markers in the breeding and conservation of M. wufengensis.
Assuntos
Conservação dos Recursos Naturais/métodos , Marcadores Genéticos/genética , Variação Genética , Magnolia/genética , Repetições de Microssatélites/genética , Melhoramento Vegetal/métodos , Análise de Variância , Antocianinas/análise , Antocianinas/metabolismo , China , Cromatografia Líquida de Alta Pressão , Análise por Conglomerados , Cor , Espécies em Perigo de Extinção , Flores/genética , Flores/metabolismo , Magnolia/classificação , Magnolia/metabolismo , Fenótipo , Filogenia , Pigmentação/genética , Especificidade da EspécieRESUMO
Stem cells from human exfoliated deciduous teeth (SHEDs) have great potential to treat various dental-related diseases in regenerative medicine. They are usually maintained with 10% fetal bovine serum (FBS) in vitro. Modified platelet-rich plasma (mPRP) would be a safe alternative to 10% FBS during SHEDs culture. Therefore, our study aimed to compare the proliferation and differentiation of SHEDs cultured in mPRP and FBS medium to explore an optimal concentration of mPRP for SHEDs maintenance. Platelets were harvested by automatic blood cell analyzer and activated by repeated liquid nitrogen freezing and thawing. The platelet-related cytokines were examined and analyzed by ELISA. SHEDs were extracted and cultured with different concentrations of mPRP or 10% FBS medium. Alkaline phosphatase (ALP) activity was measured. Mineralization factors, RUNX2 and OCN, were measured by real-time PCR. SHEDs were characterized with mesenchymal stem cells (MSCs) markers including vimentin, CD44, and CD105. mPRP at different concentrations (2, 5, 10, and 20%) enhanced the growth of SHEDs. Moreover, mPRP significantly stimulated ALP activity and promoted expression of RUNX2 and OCN compared with 10% FBS. mPRP could efficiently facilitate proliferation and differentiation of SHEDs, and 2% mPRP would be an optimal substitute for 10% FBS during SHEDs expansion and differentiation in clinical scale manufacturing.
Assuntos
Proliferação de Células , Polpa Dentária/citologia , Células-Tronco Mesenquimais/citologia , Plasma Rico em Plaquetas , Dente Decíduo/citologia , Fosfatase Alcalina/antagonistas & inibidores , Análise de Variância , Animais , Bovinos , Técnicas de Cultura de Células/métodos , Diferenciação Celular/fisiologia , Proliferação de Células/fisiologia , Células Cultivadas , Subunidade alfa 1 de Fator de Ligação ao Core/análise , Meios de Cultura , Ensaio de Imunoadsorção Enzimática , Humanos , Fator de Crescimento Derivado de Plaquetas/análise , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Fatores de Tempo , Fator de Crescimento Transformador beta1/análiseRESUMO
The objective of this study was to examine the relationship between the expression of B cell activating factor (BAFF) and BAFF receptor in patients with disease activity of systemic lupus erythematosus (SLE). Real-time RT-PCR was used to examine BAFF mRNA expression in peripheral blood monocytes of active and stable SLE patients and healthy controls. The percentage of BAFF receptor 3 (BR3) on B lymphocytes was measured by flow cytometry. Soluble BAFF levels in serum were assayed by ELISA. Microalbumin levels were assayed by an automatic immune analysis machine. BAFF mRNA and soluble BAFF levels were highest in the active SLE group, followed by the stable SLE group, and controls (P<0.01). The percentage of BR3 on B lymphocytes was downregulated in the active SLE group compared with the stable SLE group and controls (P<0.01). BAFF mRNA levels and soluble BAFF levels were higher in patients who were positive for proteinuria than in those who were negative (P<0.01). The percentage of BR3 on B lymphocytes was lower in patients who were positive for proteinuria than in those who were negative (P<0.01). The BAFF/BR3 axis may be over-activated in SLE patients. BAFF and BR3 levels may be useful parameters for evaluating treatment.
Assuntos
Humanos , Masculino , Feminino , Adolescente , Adulto , Pessoa de Meia-Idade , Adulto Jovem , Fator Ativador de Células B/metabolismo , Receptor do Fator Ativador de Células B/metabolismo , Lúpus Eritematoso Sistêmico/metabolismo , Albuminúria/urina , Fator Ativador de Células B/análise , Fator Ativador de Células B/genética , Receptor do Fator Ativador de Células B/análise , Receptor do Fator Ativador de Células B/genética , Linfócitos B/metabolismo , Biomarcadores/metabolismo , Leucócitos Mononucleares/metabolismo , Lúpus Eritematoso Sistêmico/imunologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
The objective of this study was to examine the relationship between the expression of B cell activating factor (BAFF) and BAFF receptor in patients with disease activity of systemic lupus erythematosus (SLE). Real-time RT-PCR was used to examine BAFF mRNA expression in peripheral blood monocytes of active and stable SLE patients and healthy controls. The percentage of BAFF receptor 3 (BR3) on B lymphocytes was measured by flow cytometry. Soluble BAFF levels in serum were assayed by ELISA. Microalbumin levels were assayed by an automatic immune analysis machine. BAFF mRNA and soluble BAFF levels were highest in the active SLE group, followed by the stable SLE group, and controls (P<0.01). The percentage of BR3 on B lymphocytes was downregulated in the active SLE group compared with the stable SLE group and controls (P<0.01). BAFF mRNA levels and soluble BAFF levels were higher in patients who were positive for proteinuria than in those who were negative (P<0.01). The percentage of BR3 on B lymphocytes was lower in patients who were positive for proteinuria than in those who were negative (P<0.01). The BAFF/BR3 axis may be over-activated in SLE patients. BAFF and BR3 levels may be useful parameters for evaluating treatment.
Assuntos
Fator Ativador de Células B/metabolismo , Receptor do Fator Ativador de Células B/metabolismo , Lúpus Eritematoso Sistêmico/metabolismo , Adolescente , Adulto , Albuminúria/urina , Fator Ativador de Células B/análise , Fator Ativador de Células B/genética , Receptor do Fator Ativador de Células B/análise , Receptor do Fator Ativador de Células B/genética , Linfócitos B/metabolismo , Biomarcadores/metabolismo , Feminino , Humanos , Leucócitos Mononucleares/metabolismo , Lúpus Eritematoso Sistêmico/imunologia , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Adulto JovemRESUMO
RNA-Seq technology has been widely applied to transcriptomics, genomics, molecular marker development, and functional gene studies. In the genome, microsatellites are simple sequence repeats (SSR) with a high degree of polymorphism that are used as DNA markers in many molecular genetic studies. Using traditional methods such as magnetic bead enrichment, only a few microsatellite markers have been isolated. Coilia nasus is an anadromous, small-to-moderately sized fish species that is famous as an important fishery resource. Here, we have identified a large number of microsatellites from the fish brains by using Illumina sequencing. About 20 million Illumina reads were assembled into 148,845 unigenes. A total of 13,038 SSR motifs were identified via analysis of 3,958,293,117 (3.96 Gb) nucleotides to produce a comprehensive transcript dataset for the C. nasus brain, including mono-, di-, tri-, tetra-, and penta-repeat motifs. The most abundant type of repeat motif was di-nucleotide (42.97%), followed by mono-nucleotide (38.86%), tri-nucleotide (16.21%), tetra-nucleotide (1.83%), and penta-nucleotide (0.05%) repeat units, which is similar to the results obtained in studies in other species. These data provide a base of sequence information to improve molecular-assisted markers to study C. nasus genetic diversity.
Assuntos
Peixes/genética , Análise de Sequência de RNA/métodos , Animais , Etiquetas de Sequências Expressas , Perfilação da Expressão Gênica/métodos , Marcadores Genéticos/genética , Variação Genética , Genoma , Repetições de Microssatélites , Anotação de Sequência Molecular , Polimorfismo Genético , TranscriptomaRESUMO
PURPOSE: Human epithelial growth factor receptor 2 (HER2) is over-expressed in several malignancies and represents an important therapeutic target. Aptamers are oligonucleotides that may potentially serve as tumor-homing ligand with excellent affinity and specificity for targeted cancer therapy. However, aptamers need to have nuclease resistance in order to function in vivo. The aim of this study was to generate a novel HER2 thioaptamer with enhanced nuclease resistance. METHODS: The HER2 thioaptamer is selected in an evolutionary process called systematic evolution of ligands by exponential enrichment. RESULTS: The thioaptamer could bind to the extracellular domain of HER2 with a K d of 172 nM and had minimal cross reactivity to trypsin or IgG. Moreover, the thioaptamer was found capable of binding with the HER2-positive breast cancer cells SK-BR-3 and MDA-MB-453, but not the HER2-negative cells MDA-MB-231. Notably, the thioaptamer HY6 largely maintained its structural integrity facing the nucleases in serum, while regular DNA aptamers were mostly digested. Additionally, the thioaptamer retained the capability of binding with the HER2-positive cells in the presence of serum, whereas non-thionated HER2 aptamer lost the binding function. CONCLUSION: The results indicated that the selected thioaptamer was more resistant to nuclease than regular DNA aptamers and might potentially function as a HER2-targeting ligand in complicated environment.
Assuntos
Antineoplásicos/administração & dosagem , Aptâmeros de Nucleotídeos/administração & dosagem , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Receptor ErbB-2/antagonistas & inibidores , Neoplasias da Mama/patologia , Feminino , Citometria de Fluxo , Humanos , Técnica de Seleção de Aptâmeros , Células Tumorais CultivadasRESUMO
We examined the ability of mifepristone to reverse the in vitro drug resistance of human cervical cancer cells resistant to mitomycin-C (HeLa/MMC) cells and investigated the mechanism of this effect. A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to detect the drug resistance of HeLa/MMC cells and the reversed drug resistance in vitro. Expression levels of B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and glucosylceramide synthase (GCS) were measured in HeLa and HeLa/MMC cells. The resistance index of HeLa/MMC cells on MMC was reduced from 5.02 to 1.46 after 10 mg/mL mifepristone exposure. A combination of mifepristone upregulated the Bax/Bcl-2 protein expression ratio and apoptosis in HeLa/MMC cells. GCS expression was significantly higher in HeLa/MMC cells than in HeLa cells (P < 0.01), but distinctly declined in both cell lines after mifepristone application (P < 0.01). Mifepristone reversed the resistance of HeLa/MMC cells to MMC in vitro; the overexpression of the GCS gene and the increased expression of apoptosis-related protein Bcl-2 may play important roles in the formation of multidrug resistance in cervical cancer.
Assuntos
Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Mifepristona/farmacologia , Neoplasias do Colo do Útero/patologia , Feminino , Glucosiltransferases/genética , Células HeLa , Humanos , Proteínas Proto-Oncogênicas c-bcl-2/genéticaRESUMO
The aim of this study was to evaluate effective treatment methods of malignant pleural mesothelioma (MPM). Twenty-three patients with MPM were cured by pleural infusion chemotherapy after surgery. Median survival time and the 1-, 2-, and 3-year survival rates were analyzed on the basis of follow-up. Median survival time of all patients was 15 months (range: 3 to 89 months); the 1-, 2-, and 3-year survival rates were 69.6, 43.5, and 13.0%, respectively. The 1-year survival rates of patients in stages I, II, and III were 83.3, 62.5, and 33.3%, respectively, the 2-year survival rates were 50, 37.5, and 33.3%, respectively, and the 3-year survival rates were 34, 23, and 0%, respectively. Surgery-oriented comprehensive treatment was adopted for MPM, which could improve the prognosis and life quality of patients to some extent.
Assuntos
Antineoplásicos/uso terapêutico , Carboplatina/uso terapêutico , Neoplasias Pulmonares/tratamento farmacológico , Mesotelioma/tratamento farmacológico , Paclitaxel/uso terapêutico , Adolescente , Adulto , Idoso , Antineoplásicos/administração & dosagem , Carboplatina/administração & dosagem , Intervalo Livre de Doença , Feminino , Seguimentos , Humanos , Infusões Parenterais , Neoplasias Pulmonares/cirurgia , Masculino , Mesotelioma/cirurgia , Mesotelioma Maligno , Pessoa de Meia-Idade , Paclitaxel/administração & dosagem , Período Pós-OperatórioRESUMO
Paulownia kawakamii is a fast-growing timber tree. In this study, 21 primer sets were developed using an enriched genomic library. The genetic diversity was measured in one P. kawakamii population. The number of alleles per locus ranged from 2 to 19. The observed and expected heterozygosities varied from 0.158 to 0.842 (mean = 0.421) and from 0.376 to 0.952 (mean = 0.771), respectively. All 21 loci were also polymorphic in closely related species (P. tomentosa, P. elongata, and P. fortunei). The described markers will be useful in future population genetic studies and molecular breeding of these Paulownia species.