RESUMO
Diabetic retinopathy (DR) is a severe complication of diabetes; however, its mechanism is not fully understood. Evidence has recently revealed that long non-coding RNAs (lncRNAs) are abnormally expressed in DR, and lncRNAs may function as pivotal regulators. LncRNAs are able to modulate gene expression at the epigenetic level by acting as scaffolds of histone modification complexes and sponges of binding with microRNAs (miRNAs). LncRNAs are believed to be important epigenetic regulators, which may become beneficial in the diagnosis and therapy of DR. However, the mechanisms of lncRNAs in DR are still unclear. In this review, we summarize the possible functions and mechanisms of lncRNAs in epigenetic regulation to target genes in the progression of DR.
RESUMO
Probucol has antioxidant effects and inhibits inflammation. Farnesoid X receptor (FXR) is a nuclear receptor that regulates autophagy, which is regarded as the key cause of the activation of hepatic stellate cell (HSC). In this study, the effects of probucol on HSC activation and autophagy in vitro and vivo and the role of FXR in this progress were investigated. Results showed that probucol ameliorated hepatic fibrosis and autophagy, and increased the expression of FXR in liver in a mouse model of fibrosis induced by CCl4. And probucol could alleviate lipopolysaccharide-induced autophagy and HSC activation in vitro. In addition, probucol increased FXR expression, and the Z-guggulsterone, an antagonist of FXR, could block the effects of probucol on HSC activation and autophagy. Additionally, agonists of FXR could suppress LPS-induced autophagy and activation. These results suggest that probucol could ameliorate hepatic fibrosis, and inhibit HSC autophagy and activation, and these effects are associated with FXR.
Assuntos
Antioxidantes/farmacologia , Células Estreladas do Fígado/efeitos dos fármacos , Probucol/farmacologia , Receptores Citoplasmáticos e Nucleares/metabolismo , Animais , Autofagia , Linhagem Celular , Células Cultivadas , Células Estreladas do Fígado/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Cirrose Hepática/tratamento farmacológico , Cirrose Hepática/patologia , Camundongos Endogâmicos C57BL , RatosRESUMO
The formation of complex coacervation using chitosan and octenyl succinic anhydride modified starch (OSA starch) and microencapsulation of algae oil were investigated in this study. The zeta-potential, turbidity and coacervate yield were evaluated as a function of pH and the chitosan- OSA starch mass ratio. The highest coacervate yield was achieved at pH 6.0 with a chitosan to OSA starch ratio of 1:3 (w/w). Isothermal titration calorimetry (ITC) indicated favorable affinity (Ka = 1.51 × 105 M-1) between chitosan and OSA starch. The microcapsules yielded an encapsulation efficiency (EE) in the range of 42.8 ± 0.8%- 93.1 ± 1.2%, the loading capacity ranged between 30.4 ± 2.7% and 58.3 ± 1.3%. Fourier transform infrared spectroscopy (FT-IR) spectra and scanning electron microscopy (SEM) further confirmed the microencapsulation. In comparison with the bulk oil, the microencapsulated algae oil exhibited improved oxidative stability during storage.
Assuntos
Quitosana/química , Óleos/química , Amido/química , Fenômenos Químicos , Composição de Medicamentos , Concentração de Íons de Hidrogênio , Oxirredução , Espectroscopia de Infravermelho com Transformada de Fourier , TermodinâmicaRESUMO
Objective: To investigate the application effect of bladder function training combined with Kangaiping pills on permanent bladder stoma after radical prostatectomy (RP). Methods: The clinical data of 80 patients with a permanent bladder stoma after RP in our hospital from December 2018 to December 2019 were retrospectively analyzed, and they were equally split into the experimental group (EG) and control group (CG) according to the odd and even hospitalization numbers. EG received bladder function training combined with Kangaiping pills while CG received routine nursing for permanent bladder stomas to compare the urodynamic indexes and quality of life (QOL) scores after intervention between the two groups. Results: Compared with CG, EG after intervention achieved an obviously higher number of patients with bladder function grade I (∗), higher urodynamic indexes (P < 0.001), a higher SF-36 score (P < 0.001), a lower LUTS score (P < 0.001), and a lower total incidence of postoperative adverse reactions (P < 0.05). Conclusion: Bladder function training combined with Kangaiping pills is a reliable method to improve the bladder function of patients with a permanent bladder stoma after RP. This intervention method greatly enhances the QOL of patients and reduces the risk of postoperative adverse reactions, which is recommended for clinical application.
RESUMO
BACKGROUND: The development of Cancer Stem-like Cells (CSCs) is one of the main causes of ovarian cancer tolerance to radiotherapy. Autophagy is an adaptive process by which cells damage due to radiation. As a metabolite of riboflavin, lumiflavin can enhance the chemotherapeutic effects of cisplatin on ovarian cancer CSCs. OBJECTIVE: This study aimed to investigate the synergistic effects of lumiflavin and ionising radiation on ovarian cancer CSCs and explore the association of this metabolite with autophagy. METHODS: CSCs of human ovarian cancer cell lines HO8910 were treated with lumiflavin and rapamycin and then subjected to irradiation at a cumulative dose of 8 Gy. Cell proliferation ability, clonal formation ability, apoptosis rate, autophagy changes and autophagy-related protein changes were detected. RESULTS: Lumiflavin and ionising radiation synergistically reduced cell vitality and clone formation and increased the apoptosis of CSCs compared with irradiation alone. In addition, ionising radiation increased autophagy and the expression of associated proteins, whereas lumiflavin reduced those changes in autophagy progression. Moreover, rapamycin, an autophagy inhibitor, was observed to block the synergistic effects of lumiflavin and ionising radiation on CSC apoptosis. CONCLUSION: Lumiflavin can enhance the effects of ionising radiation on ovarian cancer CSCs. The mechanism by which these effects are exerted is related to blocking the autophagy pathway.
Assuntos
Antineoplásicos/farmacologia , Autofagia/efeitos dos fármacos , Flavinas/farmacologia , Células-Tronco Neoplásicas/efeitos dos fármacos , Neoplasias Ovarianas/tratamento farmacológico , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Humanos , Neoplasias Ovarianas/patologia , Neoplasias Ovarianas/radioterapia , Radiação Ionizante , Células Tumorais CultivadasRESUMO
BACKGROUND: Bladder cancer is a major urinary system cancer, and its mechanism of action regarding its progression is unclear. The goal of this study was to examine the expression of ADAM panel in the clinical specimens of bladder cancer and to investigate the role of miR-3174/ADAM15 (a disintegrin and metalloprotease 15) axis in the regulation of bladder cancer cell proliferation. METHODS: The expression of an ADAM gene panel (including ADAM8, 9, 10, 11, 12, 15, 17, 19, 22, 23, 28, and 33), including 30 pairs of bladder tumor and non-tumor specimens, was examined by Ion AmpliSeq Targeted Sequencing. A microRNA (miRNA) that could potentially target the ADAM with the highest expression level in the tumor tissue was identified using the online tool miRDB. Next, the interaction between the miRNA and ADAM15 was identified by Western blot. Finally, the proliferation of bladder cancer cells was examined using MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) experiments (cell proliferation examining) and subcutaneous tumor models by using nude mice. RESULTS: The expression of ADAM15 in tumor tissue was found statistically significant when compared to its expression in non-tumor tissue. Additionally, ADAM15's expression in tumor tissue was found the highest of all other tested ADAMs. Next, by using the online tool miRDB, a microRNA termed miR-3174 was identified that targets ADAM15 and inhibits its expression by binding to its 3'-untranslated region. Finally, we found that overexpression of miR-3174 in bladder cancer cells inhibited the proliferation of cells due to the inhibition of ADAM15. CONCLUSION: In the present work, the data highlight that miR-3174 inhibits the proliferation of bladder cancer cells by targeting ADAM15.
RESUMO
BACKGROUND AND AIMS: Cardiac fibrosis after myocardial infarction (MI) is involved in fibroblast transforming and differentiating into myofibroblast phenoconversion, however, the underlying mechanisms are poorly understood. Toll-like receptor 4 (TLR4)-mediated pathogen-associated molecular patterns are key factors that deteriorate cardiac remodelling after MI. Moreover, autophagy has dual roles in cell survival in myocardial tissues after MI. We evaluated the relationship between TLR4 signalling and cardiac myofibroblast transformation-differentiation after MI in vivo and in vitro and analysed the role of autophagy. METHODS: We reproduced a model of MI by the permanent ligation of the left anterior descending coronary artery of Tlr4-knockout (Tlr4-/-) and wild-type (WT) male mice. We evaluated scar formation, myofibroblast phenoconversion, LC3 dot formation, autophagy related proteins and α-smooth muscle actin (SMA) in cardiac tissues, 7, 14, and 28 days after myocardial infarction. Cardiac fibroblasts were cultured from Tlr4-/- or WT mice. Vimentin, α-SMA, bilayer membrane vesicle structures of autophagosomes, and autophagy related proteins were observed after treatment with lipopolysaccharide (LPS) or 3-methyladenine (3-MA) at 24â¯h. RESULTS: After MI on 7, 14, and 28 days, Tlr4-/- mice showed that heart tissue fibrosis and expression of α-SMA, a marker of myofibroblasts, were decreased compared to WT mice. Additionally, levels of LC3II, Atg5, Atg7, and Beclin-1, which are involved in autophagy, were lower than those in WT mice. Further, p62 expression, which is negatively correlated with autophagy levels, was higher in Tlr4-/- mice. Moreover, LC3-labelled autophagosomes in cardiac tissues were reduced in these animals. In vitro, LPS, a ligand of TLR4, stimulated α-SMA expression in cardiac fibroblasts, enhanced autophagic flux, and increased autophagosome numbers. In contrast, these effects were not obvious in Tlr4-/- cardiac fibroblasts. LC3II, Atg5, Atg7, and Beclin-1 were upregulated, and p62 was downregulated in cardiac fibroblasts of WT mice stimulated with LPS. However, these effects were blocked by 3-methyladenine, an inhibitor of autophagy. CONCLUSIONS: These results suggest that TLR4 signalling executes the development of a myofibroblast phenotype after MI via autophagy and could be therapeutically exploited to improve outcome after myocardial injury.
Assuntos
Autofagossomos/metabolismo , Autofagia , Infarto do Miocárdio/metabolismo , Miocárdio/metabolismo , Miofibroblastos/metabolismo , Receptor 4 Toll-Like/metabolismo , Animais , Autofagossomos/patologia , Células Cultivadas , Modelos Animais de Doenças , Fibrose , Masculino , Camundongos Endogâmicos C3H , Camundongos Knockout , Infarto do Miocárdio/patologia , Miocárdio/patologia , Miofibroblastos/patologia , Fenótipo , Transdução de Sinais , Receptor 4 Toll-Like/deficiência , Receptor 4 Toll-Like/genéticaRESUMO
INI1/hSNF5/BAF47 is a core component of the hSWI/ SNF ATP-dependent chromatin remodeling complex, and it has been implicated in regulating gene expression, cell division and tumorigenesis. We investigated whether INI1/hSNF5/BAF47 functions in activation of the colony stimulating factor 1 (CSF1) promoter in HeLa cells. Overexpression of INI1/hSNF5/BAF47 promoted CSF1 transcription, and siRNA targeting INI1/hSNF5/ BAF47 (siINI1) strongly inhibited the activity of the CSF1 promoter. We demonstrated that all conserved domains of INI1/hSNF5/BAF47 are needed for CSF1 transcription. ChIP experiment showed that INI1/ hSNF5/BAF47 is recruited to the region of the CSF1 promoter. Taken together, these results indicate that INI1/hSNF5/BAF47 is involved in activation of the CSF1 promoter.
Assuntos
Proteínas de Ligação a DNA/fisiologia , Regulação da Expressão Gênica/fisiologia , Fator Estimulador de Colônias de Macrófagos/genética , Proteínas Repressoras/metabolismo , Fatores de Transcrição/fisiologia , Técnicas de Cultura de Células , Montagem e Desmontagem da Cromatina/genética , Proteínas Cromossômicas não Histona , Proteínas de Ligação a DNA/antagonistas & inibidores , Proteínas de Ligação a DNA/genética , Células HeLa , Humanos , Fator Estimulador de Colônias de Macrófagos/biossíntese , Plasmídeos/genética , Regiões Promotoras Genéticas , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , RNA Interferente Pequeno/genética , Proteínas Repressoras/genética , Proteína SMARCB1 , Fatores de Transcrição/antagonistas & inibidores , Fatores de Transcrição/genética , Transcrição Gênica/fisiologiaRESUMO
CrV0.95P0.05O4 prepared as a pure crystalline form was found to be highly active for the vapor-phase oxidation of picolines to the corresponding aldehydes and acids in the presence of water.
RESUMO
Degenerate PCR is introduced in this paper, including what is degenerate PCR, how to design degenerate primers, how to optimize degenerate PCR parameters, how to applying degenerate PCR to obtain full-length gene and which fields can apply degenerate PCR. The limits and recent advances of degenerate PCR are also discussed. Based on this introduction,strategies of gene cloning and applications of degenerate PCR in gene cloning are summarized in brief. Degenerate PCR is a very useful tool for searching and discovering new genes and new members of a protein family.
RESUMO
A new strain FS3 which could achieve an efficient bioconversion of raw corn stalk to hydrogen had been isolated from anaerobic acclimated sludge, and identified as Clostridium butyricum on the basis of a series of physiological and biochemical experiments and 16S rDNA gene sequence. The strain could utilize various carbon sources to produce hydrogen. On the basis of single-factor experiments, the response surface methodology (RSM) was performed to optimize the media for hydrogen production. The maximum hydrogen yield of 92.9ml/g was observed under the optimal conditions: 20g/l raw corn stalk, 1.76g/l NH4HCO3, 0.91g/l KH2PO4 and 10.4ml/l nutrient solution. This finding opens a new avenue for direct conversion of raw cellulosic biomass to bio-hydrogen.
Assuntos
Clostridium/metabolismo , Hidrogênio/metabolismo , Resíduos , Zea mays/química , Bicarbonatos/farmacologia , Biomassa , Carbono/farmacologia , Celulose/metabolismo , Clostridium/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Fosfatos/farmacologia , Compostos de Potássio/farmacologia , Análise de Regressão , SoluçõesRESUMO
Actin is an important protein in nucleus and has been implicated in transcription, however, the mechanism of its function in transcription is still not clear. In this article, we studied the role of actin in the regulation of human CSF1 gene transcription. Our results showed that nuclear actin stimulates the activity of CSF1 promoter, and the role in augmenting CSF1 gene transcription requires the formation of chromatin and Z-DNA structure. The ATP binding motifs of nuclear actin are essential for its function in regulating CSF1 gene transcription, and upon actin overexpression, there is an increase in the ATPase activity of nuclear proteins. Further investigation revealed that nuclear actin regulates CSF1 gene transcription in a BRG1 independent manner. Together, these original results have provided evidence for further understanding the mechanism of nuclear actin in regulating gene transcription.
Assuntos
Actinas/metabolismo , Núcleo Celular/metabolismo , Cromatina/metabolismo , DNA Helicases/metabolismo , Fator Estimulador de Colônias de Macrófagos/metabolismo , Proteínas Nucleares/metabolismo , Fatores de Transcrição/metabolismo , Transcrição Gênica , Adenosina Trifosfatases/metabolismo , Trifosfato de Adenosina/metabolismo , DNA Forma Z/metabolismo , Regulação da Expressão Gênica , Células HeLa , Humanos , Fator Estimulador de Colônias de Macrófagos/genética , Regiões Promotoras GenéticasRESUMO
A strain capable of using polyvinyl alcohol (PVA) as sole carbon source was isolated from soil samples of a textile factory. The 16S rDNA sequence analysis cell morphology, physiology and biochemistry showed that it belonged to Janthinobacterium sp. This is the first report to show that the screened Janthinobacterium sp. could degrade PVA. The optimum nutritional and environmental conditions for PVA-degrading enzyme production by Janthinobacterium sp. were investigated by single-factor tests. Under optimized nutritional and environmental condition in shake flasks, PVA-degrading enzyme reached 5.12 U/mL at 21 h. With PVA-degrading enzyme produced by Janthinobacterium sp. WSH04-01, 80% of PVA could be degraded from cotton fabrics in 3 h.
Assuntos
Oxirredutases do Álcool/química , Oxirredutases do Álcool/metabolismo , Fibra de Algodão , Álcool de Polivinil/química , Álcool de Polivinil/metabolismo , Proteobactérias/enzimologia , Indústria Têxtil/métodos , Biodegradação AmbientalRESUMO
c-Abl tyrosine kinase, predominantly distributed in nucleus, has been implicated in many important cellular processes including the regulation of gene transcription. In this study, we showed that c-Abl promoted the transcription of c-fos gene, both exogenously and endogenously. The nuclear localization and tyrosine kinase activity of c-Abl were required for the activation of c-fos gene. c-Abl was associated with RNA polymerase II (RNAP II) in vivo and augmented the tyrosine phosphorylation of the largest subunit of RNAP II. In addition, c-Abl and RNAP II could be recruited to the region of c-fos promoter. The combined results suggest that c-Abl plays an important role in the transcriptional regulation of c-fos gene and the tyrosine phosphorylation of the largest subunit of RNAP II by c-Abl is involved in the regulating process.
Assuntos
Regulação da Expressão Gênica , Genes fos/genética , Proteínas Proto-Oncogênicas c-abl/metabolismo , RNA Polimerase II/metabolismo , Transporte Ativo do Núcleo Celular , Linhagem Celular , Humanos , Células K562 , Mutação/genética , Fosforilação , Fosfotirosina/metabolismo , Regiões Promotoras Genéticas/genética , Ligação Proteica , Subunidades Proteicas/química , Subunidades Proteicas/metabolismo , Proteínas Proto-Oncogênicas c-abl/química , Proteínas Proto-Oncogênicas c-abl/genética , RNA Polimerase II/química , Transcrição Gênica/genéticaRESUMO
Nuclear actin is a nuclear component in many kinds of eukaryotic cells but its function is still not clear. In this study, we overexpressed actin in nuclei and found that it promoted transcription of the CSF-1 gene, both exogenous and endogenous, approximately two-fold. Cytochalasin B did not affect this function of nuclear actin. Our results suggest that nuclear actin plays a role in regulating CSF-1 gene transcription, and this role does not depend on actin polymerization.