RESUMO
Formation of hepatocyte spheroids is a necessary strategy for increasing liver-specific function in vitro. In this study, HepG2 cells showed good viability when grown on a polylactic acid-chitosan (PLA-CS) nanofiber and aggregated to form multicellular spheroids on the PLA-CS nanofibers with a diameter of approximately 100-200 mm in 5 days of culture, whereas no such aggregation was observed in cells cultured on 24-well plates. Hepatocyte spheroids formed on the PLA-CS nanofibers displayed excellent hepatic-related protein expression, such as albumin and urea, compared to HepG2 cells cultured on the 24-well plates. These results indicated that formation of the hepatocyte spheroids in nanofibers can increase and maintain hepatocyte functions for a longer time, supporting a new strategy for bioartificial liver development.
Assuntos
Quitosana/química , Nanofibras/química , Poliésteres/química , Esferoides Celulares/fisiologia , Albuminas/biossíntese , Albuminas/metabolismo , Órgãos Artificiais , Agregação Celular , Sobrevivência Celular , Quitosana/farmacologia , Meios de Cultura/química , Células Hep G2 , Humanos , Fígado/citologia , Tamanho da Partícula , Poliésteres/farmacologia , Esferoides Celulares/efeitos dos fármacos , Ureia/metabolismoRESUMO
Shenkangling plays a role of Yishenhuoxue effect for the treatment of children with nephrotic syndrome. The aim of this study was to investigate the effects of Shenkangling intervention on the mitogen-activated protein kinase (MAPK) pathway in rats with Adriamycin-induced nephropathy (AN) and its underlying mechanism of action. Nephrosis was induced in healthy Sprague-Dawley rats by doxorubicin and the rats were untreated or treated with prednisone, simvastatin, Shenkangling, or a combination thereof. Using real-time PCR, the mRNA expression levels of Chemokine (C-X-C motif) ligand 16 (CXCL16), A Disintegrin and metalloproteinase domain-containing protein 10 (ADAM10), and ADAM17 in the renal tissues of these rats were found to be decreased by the various treatments compared to those in the untreated doxorubicin-induced nephrosis rats. To quantify the activation of the MAPK pathway, western blotting was used to detect the phosphorylation levels of MAPK pathway-associated proteins (p38, ERK1/2, SAPK/JNK) and nuclear factor (NF)-κB p65, which were reduced by the various treatments compared to those in the untreated doxorubicin-induced rats. Serum levels of transforming growth factor (TGF)-ß1, tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, and IL-6, quantified by ELISA, were decreased by the various treatments compared to the levels in the untreated doxorubicin-induced nephrosis rats. The rats treated with prednisone, simvastatin, and Shenkangling showed the best outcome. The Chinese medicine Shenkangling that is known for nourishing the kidney and promoting blood circulation reduced urinary protein levels, increased serum albumin levels, and reduced cholesterol levels by reducing the release of CXCL16, ADAM10, ADAM17, TGF-ß1, TNF-α, IL-1ß, IL- 6, and other inflammatory mediators and inhibiting the activation of the MAPK signaling pathway, thereby effectively improving the state of nephropathy in AN rats. These results indicate that Shenkangling can be used clinically to treat nephropathy.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Síndrome Nefrótica/tratamento farmacológico , Proteína ADAM10/genética , Proteína ADAM17/genética , Animais , Quimiocina CXCL6/genética , Doxorrubicina/toxicidade , Interleucina-1beta/sangue , Interleucina-6/sangue , Masculino , NF-kappa B/metabolismo , Síndrome Nefrótica/sangue , Síndrome Nefrótica/induzido quimicamente , Síndrome Nefrótica/enzimologia , Proteinúria/tratamento farmacológico , Proteinúria/enzimologia , Proteinúria/metabolismo , Ratos , Ratos Sprague-Dawley , Fator de Crescimento Transformador beta1/sangue , Fator de Necrose Tumoral alfa/sangueRESUMO
With advances in molecular biology, microarray data have become an important resource in the exploration of complex human diseases. Although gene chip technology continues to grow, there are still many barriers to overcome, such as high costs, small sample sizes, complex procedures, poor repeatability, and the dependence on data analysis methods. To avoid these problems, simulation data have a vital role in the study of complex diseases. A simulation method of microarray data is introduced in this study to model the occurrence and development of general diseases. Using classic statistics and control theory, five risk models are proposed. One or more models can be introduced into the baseline simulation dataset with a case-control label. In addition, time-series gene expression data can be generated to model the dynamic evolutionary process of a disease. The prevalence of each model is estimated and disease-associated genes are tested by significance analysis of microarrays. The source code, written in MATLAB, is freely and publicly available at http://sourceforge.net/projects/genesimulation/files/.
Assuntos
Perfilação da Expressão Gênica/métodos , Análise em Microsséries/métodos , Algoritmos , Estudos de Casos e Controles , Simulação por Computador , Bases de Dados Genéticas , Humanos , Modelos Genéticos , Modelos Estatísticos , SoftwareRESUMO
Piwi-interacting RNAs (piRNAs) are a class of small non-coding RNAs. Distinguishing piRNAs from other non-coding RNAs is important because of their important role in the physiological regulation of spermatogenesis, genome protection from transposons, and regulation of mRNAs and long non-coding RNAs. Few computational studies have addressed piRNAs detection, and both effectiveness and efficiency of piRNA detection tools require improvement. In this study, a piRNA detection method based on sequence features and a support vector machine was developed. Four types of features are proposed: weighted k-mer, weighted k-mer with wildcards, position-specific base, and piRNA length. The piRNA sequences from human, mouse, rat, and drosophila were respectively used in this experiment. Compared to existing algorithms, the proposed method provides a better balance between precision and sensitivity (both are approximately 90%), and although these values were slightly slower than those obtained using the piRNA annotation approach, the proposed method was four-fold faster than piRPred and 229-fold faster than piRNA predictor.
Assuntos
RNA Interferente Pequeno/análise , RNA Interferente Pequeno/genética , Análise de Sequência de RNA/métodos , Algoritmos , Animais , Drosophila/genética , Genoma , Humanos , RNA Interferente Pequeno/isolamento & purificação , RNA Interferente Pequeno/metabolismo , Software , Máquina de Vetores de SuporteRESUMO
Numerous studies have evaluated the association between the Ala1330Val polymorphism of the low-density lipoprotein receptor-related proteins 5 (LRP5) gene and fracture risk. However, the specific association is still controversial. The aim of this study was to investigate their correlation via meta-analysis. and EMBASE databases were searched, and data were extracted independently by two reviewers. Odds ratios (ORs) with corresponding 95% confidence intervals (CIs) were used to assess the strength of the associations. Statistical analysis was performed using the STATA 12.0 software. Seven studies, involving 808 cases and 1586 controls, were included in the analysis. Meta-analysis results showed no significant association between the LRP5 Ala1330Val polymorphism and fracture risk (ValVal vs AlaAla: OR = 1.25, 95%CI = 0.82-1.91; AlaVal vs AlaAla: OR = 1.16, 95%CI = 0.95-1.42; dominant model: OR = 1.77, 95%CI = 0.96-1.41; recessive model: OR = 1.21, 95%CI = 0.80-1.83). Taking into account the effect of ethnicity, the LRP5 Ala1330Val polymorphism was not associated with the risk of fracture in Asians or Caucasians. The results of the current meta-analysis indicate that the LRP5 Ala1330Val polymorphism may not be correlated with fracture susceptibility.
Assuntos
Fraturas Ósseas/genética , Proteína-5 Relacionada a Receptor de Lipoproteína de Baixa Densidade/genética , Polimorfismo de Nucleotídeo Único , Povo Asiático , Estudos de Casos e Controles , Fraturas Ósseas/etnologia , Humanos , Mutação de Sentido Incorreto , População BrancaRESUMO
To investigate genetic diversity and relationships of 101 faba bean (Vicia faba L.), landraces and varieties from different provinces of China and abroad were analyzed by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE). A total of 2625 unambiguous and stable bands from 101 germplasms were detected, and 36 different bands were classified according to the electrophoretic mobility patterns of the proteins as determined by the SDS-PAGE analysis, of which 16 were polymorphic. Besides the common bands, the protein bands of 92, 75, 62, 40, 34, 17, and 13 kDa presented the highest frequencies of 92.08, 90.10, 99.01, 95.05, 95.05, 98.02, and 95.05%, respectively. The other 29 polymorphic protein bands showed higher polymorphism with 16.09 polymorphic bands in average. The genetic similarity of the 101 genotypes tested varied from 0.6111 to 0.9722, with an average of 0.7122. Cluster analysis divided the 101 genotypes into six major clusters, which was consistent with the systematic classification of faba bean done in previous studies. The overall results indicated that SDS-PAGE was a useful tool for genetic diversity analysis and laid a solid foundation for future faba bean breeding.
Assuntos
Variação Genética , Vicia faba/genética , Análise por Conglomerados , Eletroforese em Gel de Poliacrilamida , Marcadores Genéticos , Genótipo , Polimorfismo Genético , Vicia faba/classificaçãoRESUMO
The aim of this study was to optimize candidate antigen proteins for serological screening of Chlamydia trachomatis infection. C. trachomatis positive serum and swabs of genital secretions were collected from 50 patients in the Tianjin Medical University General Hospital, as well as from 30 patients negative for C. trachomatis. Samples were assessed by colloidal gold assay in a sexually transmitted disease clinic as follows: serum antibodies for eight kinds of C. trachomatis immunodominant proteins (Pgp3, CPAF, CT143, CT101, CT694, CT875, CT813, and IncA) were detected, and two traditional gold standards, immunofluorescence and C. trachomatis cell culture of genital secretions, were used for comparison in order to determine the antigen protein combinations with the highest sensitivity and specificity. Of the 50 samples that tested positive for C. trachomatis infection by colloidal gold assay, 44 tested positive by micro-immunofluorescence, whereas 6 tested negative. In contrast, 14 samples tested positive by cell culture, whereas 36 tested negative. Serological results of the immunodominant protein combination of Pgp3, CT694, and CT875 shared positive coincidence rates of 97.73 and 92.86% with C. trachomatis micro-immunofluorescence and cell culture, respectively. No antibodies of the three proteins were detected in the 30 C. trachomatis samples that tested negative by colloidal gold assay; these samples also tested negative in C. trachomatis genital secretion culture. Overall, the combination of the three immunodominant proteins Pgp3, CT694, and CT875 had good sensitivity and specificity for serological screening of C. trachomatis infection, and the process was simple and easy to apply.
Assuntos
Proteínas de Bactérias/sangue , Infecções por Chlamydia/sangue , Chlamydia trachomatis/metabolismo , Chlamydia trachomatis/química , Eletroforese em Gel de Poliacrilamida , Humanos , Reação em Cadeia da PolimeraseRESUMO
Stroke is a non-communicable disease of increasing socioeconomic importance in aging populations. This study compared the risk factors implicated in two subtypes of ischemic stroke: lacunar stroke (LS) and non-lacunar stroke (NLS). A retrospective case control study was conducted on a total of 368 patients [220 cases (59.8%) of NLS and 148 cases (40.2%) of LS] with first-time onset of ischemic stroke. Multivariate logistic regression was performed to compare multiple non-cerebrovascular risk factors between the two groups. More patients with a history of diabetes were found in the NLS than the LS group (40.5 vs 26.4%), and that both fasting glucose and HbA1C levels before the onset of stroke were higher in NLS than LS patients. Multivariate analysis revealed that patients with a history of diabetes were 1.57 times more likely to have NLS than LS (OR = 1.57, 95%CI = 0.95-3.26). Moreover, male patients were more likely to develop NLS than females (OR = 1.46, 95%CI = 0.79-2.69), and patients with elevated fibrinogen levels were 1.4 times more likely to develop NLS than LS (OR = 1.40, 95%CI = 1.09-1.80). Additionally, patients who were heavy drinkers (OR = 1.39, 95%CI = 0.68-2.84) or smokers (OR = 1.62, 95%CI = 0.91-2.89) were more likely to develop NLS than LS. Other risk factors, such as hypertension, dyslipidemia, age, and average blood pressure, did not differ between the two types of stroke. Thus, distinct non-cerebrovascular risk factors (male gender, long history of diabetes, elevated fibrinogen, heavy smoking, and heavy drinking) are associated with a higher risk of developing non-lacunar stroke than lacunar stroke.
Assuntos
Isquemia Encefálica/complicações , Acidente Vascular Cerebral Lacunar/etiologia , Acidente Vascular Cerebral/etiologia , Idoso , Consumo de Bebidas Alcoólicas/efeitos adversos , Glicemia/metabolismo , Estudos de Casos e Controles , Complicações do Diabetes/sangue , Complicações do Diabetes/etiologia , Dislipidemias/complicações , Jejum/sangue , Feminino , Fibrinogênio/metabolismo , Hemoglobinas Glicadas/metabolismo , Humanos , Hipertensão/complicações , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Estudos Retrospectivos , Fatores de Risco , Fatores Sexuais , Fumar/efeitos adversos , Acidente Vascular Cerebral/sangue , Acidente Vascular Cerebral Lacunar/sangueRESUMO
Bone morphogenetic protein-7 (BMP-7) and SOX9 are important transcription factors in chondrogenesis. In this study, we examined the biological function of the adeno-associated virus (AAV)-mediated BMP-7 and SOX9 double gene in vitro co-transfection of nucleus pulposus cells of human degenerative intervertebral disc. Human intervertebral disc nucleus pulposus cells were cultured in vitro and subcultured for 5 generations. Using rAAV-BMP-7 and rAAV-SOX9 AAV2-type AAV viruses, the cells were divided into 4 groups: blank normal, BMP-7 transfection, SOX9 transfection, and BMP-7 and SOX9 co-transfection. After 48 h, expression of type II collagen and its mRNA in nucleus pulposus cells was determined. The expression of type II collagen in BMP-7 transfection, SOX9 transfection, and co-transfection groups was up-regulated to varying degrees compared to the blank control group. The type II collagen mRNA level expression in the co-transfection group was significantly higher than in other groups (P < 0.05). AAV-mediated BMP-7 and SOX9 in vitro co-transfection can promote the synthesis of type II collagen in nucleus pulposus cells in the human degenerative intervertebral disc. Double-gene therapy has a synergistic effect in the treatment of intervertebral disc degeneration.
Assuntos
Proteína Morfogenética Óssea 7/genética , Dependovirus/genética , Terapia Genética , Degeneração do Disco Intervertebral/terapia , Fatores de Transcrição SOX9/genética , Proteína Morfogenética Óssea 7/biossíntese , Células Cultivadas , Colágeno Tipo II/biossíntese , Colágeno Tipo II/genética , Expressão Gênica , Vetores Genéticos , Proteínas de Fluorescência Verde/biossíntese , Proteínas de Fluorescência Verde/genética , Humanos , Disco Intervertebral/patologia , Degeneração do Disco Intervertebral/metabolismo , Fatores de Transcrição SOX9/biossíntese , TransfecçãoRESUMO
Fengshui forests (sacred groves) are important in traditional Chinese culture and home to many endangered species. These forests may provide protection for some endangered plant species outside the nature reserves, but little is known about their role in genetic conservation. Using inter-simple sequence repeat (ISSR) markers, we compared the genetic diversity of 6 populations of Phoebe bournei (Hemsl.) Yang, a commercially important woody species, which is under second-class national protection and endemic to China. Samples were collected from the nature reserves and Fengshui forests in southern China. Herein, we show that Fengshui forest populations are capable of maintaining some level of genetic diversity. For nature reserve populations, the average NA and NE were 1.58 and 1.39, respectively; and for Fengshui forests, they were 1.39 and 1.12, respectively. For nature reserve populations, Nei's gene diversity (H) and Shannon's index (I) were 0.32 and 0.11, respectively; and for Fengshui forests, they were 0.22 and 0.07, respectively. We discuss the reasons for the genetic differences between populations of the Fengshui forests and nature reserves and propose conservation strategies for the Fengshui forest.