RESUMO
Lotus plumule, the embryo of the seed of the sacred lotus (Nelumbo nucifera), contains a high accumulation of secondary metabolites including flavonoids and possesses important pharmaceutical value. Flavonoid C-glycosides, which accumulate exclusively in lotus plumule, have attracted considerable attention in recent decades due to their unique chemical structure and special bioactivities. As well as mono-C-glycosides, lotus plumule also accumulates various kinds of di-C-glycosides by mechanisms which are as yet unclear. In this study we identified two C-glycosyltransferase (CGT) genes by mining sacred lotus genome data and provide in vitro and in planta evidence that these two enzymes (NnCGT1 and NnCGT2, also designated as UGT708N1 and UGT708N2, respectively) exhibit CGT activity. Recombinant UGT708N1 and UGT708N2 can C-glycosylate 2-hydroxyflavanones and 2-hydroxynaringenin C-glucoside, forming flavone mono-C-glycosides and di-C-glycosides, respectively, after dehydration. In addition, the above reactions were successfully catalysed by cell-free extracts from tobacco leaves transiently expressing NnCGT1 or NnCGT2. Finally, enzyme assays using cell-free extracts of lotus plumule suggested that flavone di-C-glycosides (vicenin-1, vicenin-3, schaftoside and isoschaftoside) are biosynthesized through sequentially C-glucosylating and C-arabinosylating/C-xylosylating 2-hydroxynaringenin. Taken together, our results provide novel insights into the biosynthesis of flavonoid di-C-glycosides by proposing a new biosynthetic pathway for flavone C-glycosides in N. nucifera and identifying a novel uridine diphosphate-glycosyltransferase (UGT708N2) that specifically catalyses the second glycsosylation, C-arabinosylating and C-xylosylating 2-hydroxynaringenin C-glucoside.
Assuntos
Flavonoides/metabolismo , Glicosídeos/metabolismo , Nelumbo/metabolismo , Glicosilação , Glicosiltransferases/genética , Glicosiltransferases/metabolismo , Redes e Vias Metabólicas , Nelumbo/enzimologia , Nelumbo/genética , Filogenia , Plantas Geneticamente Modificadas , NicotianaRESUMO
Ligustrazine was the active ingredient of the traditional Chinese medicine Chuanxiong Rhizoma. However, the content of ligustrazine is very low. We proposed a hypothesis that ligustrazine was produced by the mutual effects between endophytic Bacillus subtilis and the Ligusticum chuanxiong Hort. This study aimed to explore whether the endophytic B. subtilis LB5 could make use of Chuanxiong Rhizoma fermentation matrix to produce ligustrazine and clarify the mechanisms of action preliminarily. Ultra high performance liquid chromatography with quadrupole time-of-flight mass spectrometry analysis showed the content of ligustrazine in Chuanxiong Rhizoma was below the detection limit (0.1 ng/mL), while B. subtilis LB5 produced ligustrazine at the yield of 1.0268 mg/mL in the Chuanxiong Rhizoma-ammonium sulfate fermentation medium. In the fermented matrix, the reducing sugar had a significant reduction from 12.034 to 2.424 mg/mL, and rough protein content increased from 2.239 to 4.361 mg/mL. Acetoin, the biosynthetic precursor of ligustrazine, was generated in the Chuanxiong Rhizoma-Ammonium sulfate (151.2 mg/mL) fermentation medium. This result showed that the endophytic bacteria B. subtilis LB5 metabolized Chuanxiong Rhizoma via secreted protein to consume the sugar in Chuanxiong Rhizoma to produce a considerable amount of ligustrazine. Collectively, our preliminary research suggested that ligustrazine was the interaction product of endophyte, but not the secondary metabolite of Chuanxiong Rhizoma itself.
Assuntos
Bacillus subtilis/química , Medicamentos de Ervas Chinesas/análise , Pirazinas/análise , Rizoma/química , Bacillus subtilis/metabolismo , Cromatografia Líquida de Alta Pressão , Medicamentos de Ervas Chinesas/metabolismo , Fermentação , Espectrometria de Massas , Medicina Tradicional Chinesa , Pirazinas/metabolismo , Rizoma/metabolismo , Fatores de TempoRESUMO
Elm fruits were once an important food source in the years of famine. Research on the functional compounds in elm fruits was almost unavailable. In this study, we established an efficient high-performance liquid chromatography method for the simultaneous separation of eight chlorogenic acids and 28 flavonoids in elm fruits for the first time. Total flavonoid contents ranged from 286 mg/100 g (Ulmus laciniata) to 1228 mg/100 g (U. pumila). High concentrations of rutin, quercetin 3-O-glucoside, and kaempferol derivatives were present in U. laevis, U. castaneifolia, and U. pumila, respectively. Furthermore, the fruit extracts of U. americana, U. castaneifolia, U. davidiana, and U. pumila showed higher antioxidant activity. These results suggest that fruits of these species can be used as bioresources for the extraction of the corresponding functional compounds. This work provides informative data and can be an important reference for future research on elm fruits as a renewed food resource.
Assuntos
Antioxidantes/análise , Ácido Clorogênico/análise , Flavonoides/análise , Frutas/química , Ulmus/química , Antioxidantes/farmacologia , Benzotiazóis/antagonistas & inibidores , Compostos de Bifenilo/antagonistas & inibidores , Ácido Clorogênico/farmacologia , Flavonoides/farmacologia , Picratos/antagonistas & inibidores , Ácidos Sulfônicos/antagonistas & inibidoresRESUMO
Ligustrazine is an important active ingredient of the traditional Chinese medicine Chuanxiong Rhizoma, but its content is a controversial topic. The endophytes of medicinal plants have the ability to produce the same active substances as the host, so this report focused on the endophytic Bacillus subtilis, to study the origin of ligustrazine in Chuanxiong Rhizoma preliminarily by inoculating the isolated endophytic B. subtilis to the Chuanxiong Rhizoma medium in vitro for solid state fermentation. Tissue grinding method was used to isolate the endogenetic B. subtilis. The morphological features, conventional physiological and biochemical reactions and 16S rRNA molecular techniques were combined to identify the endogenetic strains. Then, the strains that grew well in the medicinal matrix of Chuanxiong Rhizoma were screened out for further fermentation studies. The solid-state fermentation was performed at 37 °C for 30 d using Chuanxiong Rhizoma fermentation medium (40 g Chuanxiong Rhizoma powder, 100 mL sterile water, 121 °C, sterilization for 25 minutes). UPLC was used to detect the contents of ligustrazine, acetoin in the Chuanxiong Rhizoma fermentation medium and Chuanxiong Rhizoma. All the five strains were Gram-positive and had spores. Phylogenetic analysis of the 16S rRNA sequence showed that the endophytes were B. subtilis. The results of UPLC showed that ligustrazine was detected in the Chuanxiong Rhizoma fermentation medium inoculated with endogenetic B. subtilis LB3, LB3-2-1, LB4, LB5 and LB6-2, while not detected neither in blank Chuanxiong Rhizoma fermentation medium nor in Chuanxiong Rhizoma. This study showed that the endogenetic B. subtilis of Ligusticum chuanxiong Hort. can make use of Chuanxiong Rhizoma fermentation medium to produce ligustrazine. Endogenetic B. subtilis has a certain correlation with the accumulation of ligustrazine in Rhizoma Chuanxiong. We speculate that the ligustrazine may be derived from the catabolism of endogenetic B. subtilis in Ligusticum chuanxiong.
Assuntos
Bacillus subtilis , Ligusticum/química , Ligusticum/microbiologia , Pirazinas/análise , Endófitos , Fermentação , Filogenia , RNA Ribossômico 16S , Rizoma/químicaRESUMO
Small cell neuroendocrine carcinoma (SCNEC) of the tongue is very rare. We here present a SCNEC impatient with distant metastasis. A 74-year-old Chinese male went to hospital to treat a tongue tumor, which was founded at a conventional physical examination in Weifang Stomatology Hospital. The check of positron emission tomography-computer tomography (PET-CT) by Weifang people's hospital revealed a tumor in the right root of tongue, and distant metastasis in the right submandibular area, neck, mediastinum, right hilar, abdominal, retroperitoneal multiple lymph nodes, left thyroid, right lower lung, right scapula and bilateral adrenal. The patient was diagnosed tongue SCNEC by the pathological analysis of the tissue section. Conforming to the diagnosis of tongue SCNEC, the patient received adjuvant chemotherapy for 6 cycles with etoposide and carboplatin, and is alive now 9 months after the diagnosis.
Assuntos
Carboplatina/uso terapêutico , Carcinoma de Células Pequenas/diagnóstico por imagem , Carcinoma de Células Pequenas/tratamento farmacológico , Etoposídeo/uso terapêutico , Neoplasias da Língua/tratamento farmacológico , Idoso , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Carcinoma de Células Pequenas/patologia , Humanos , Masculino , Metástase Neoplásica/diagnóstico por imagem , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada , Neoplasias da Língua/diagnóstico por imagem , Neoplasias da Língua/patologiaRESUMO
BACKGROUND: Waterlily (Nymphaea spp.), a perennial herbaceous aquatic plant, is divided into two ecological groups: hardy waterlily and tropical waterlily. Although the hardy waterlily has no attractive blue flower cultivar, its adaptability is stronger than tropical waterlily because it can survive a cold winter. Thus, breeding hardy waterlily with real blue flowers has become an important target for breeders. Molecular breeding may be a useful way. However, molecular studies on waterlily are limited due to the lack of sequence data. RESULTS: In this study, six cDNA libraries generated from the petals of two different coloring stages of blue tropical waterlily cultivar Nymphaea 'King of Siam' were sequenced using the Illumina HiSeq™ 2500 platform. Each library produced no less than 5.65 Gb clean reads. Subsequently, de novo assembly generated 112,485 unigenes, including 26,206 unigenes annotated to seven public protein databases. Then, 127 unigenes could be identified as putative homologues of color-related genes in other species, including 28 up-regulated and 5 down-regulated unigenes. In petals, 16 flavonoids (4 anthocyanins and 12 flavonols) were detected in different contents during the color development due to the different expression levels of color-related genes, and four flavonols were detected in waterlily for the first time. Furthermore, UA3GTs were selected as the most important candidates involved in the flavonoid metabolic pathway, UA3GTs induced blue petal color formation in Nymphaea 'King of Siam'. CONCLUSIONS: This study will improve our understanding of the molecular mechanism of blue flowers in waterlily and provide the basis for molecular breeding of blue hardy waterlily cultivars.
Assuntos
Flores/genética , Flores/metabolismo , Metaboloma , Nymphaea/genética , Nymphaea/metabolismo , Transcriptoma , Biologia Computacional/métodos , Perfilação da Expressão Gênica , Metabolômica , FenótipoRESUMO
BACKGROUND: Tree peony (Paeonia section Moutan DC.) is known for its excellent ornamental and medicinal values. In 2011, seeds from P. ostii have been identified as novel resource of α-linolenic acid (ALA) for seed oil production and development in China. However, the molecular mechanism on biosynthesis of unsaturated fatty acids in tree peony seeds remains unknown. Therefore, transcriptome data is needed to better understand the underlying mechanisms. RESULTS: In this study, lipid accumulation contents were measured using GC-MS methods across developing tree peony seeds, which exhibited an extraordinary ALA content (49.3%) in P. ostii mature seeds. Transcriptome analysis was performed using Illumina sequencing platform. A total of 144 million 100-bp paired-end reads were generated from six libraries, which identified 175,874 contigs. In the KEGG Orthology enrichment of differentially expressed genes, lipid metabolism pathways were highly represented categories. Using this data we identified 388 unigenes that may be involved in de novo fatty acid and triacylglycerol biosynthesis. In particular, three unigenes (SAD, FAD2 and FAD8) encoding fatty acid desaturase with high expression levels in the fast oil accumulation stage compared with the initial stage of seed development were identified. CONCLUSIONS: This study provides the first comprehensive genomic resources characterizing tree peony seeds gene expression at the transcriptional level. These data lay the foundation for further understanding of molecular mechanism responsible for lipid biosynthesis and the high unsaturated fatty acids (especially ALA) accumulation. Meanwhile, it provides theoretical base for potential oilseed application in the respect of n-6 to n-3 ratio for human diets and future regulation of target healthy components of oils.
Assuntos
Ácidos Graxos/metabolismo , Paeonia/genética , Paeonia/metabolismo , Sementes/genética , Sementes/metabolismo , Transcriptoma , Vias Biossintéticas , Biologia Computacional , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Sequenciamento de Nucleotídeos em Larga Escala , Metabolismo dos Lipídeos , Anotação de Sequência Molecular , FenótipoRESUMO
BACKGROUND: Mounting evidence indicates that long noncoding RNAs (lncRNAs) could play a pivotal role in cancer biology. However, the overall biological role and clinical significance of PVT1 in gastric carcinogenesis remains largely unknown. METHODS: Expression of PVT1 was analyzed in 80 GC tissues and cell lines by qRT-PCR. The effect of PVT1 on proliferation was evaluated by MTT and colony formation assays, and cell apoptosis was evaluated by Flow-cytometric analysis. GC cells transfected with shPVT1 were injected into nude mice to study the effect of PVT1 on tumorigenesis in vivo. RIP was performed to confirm the interaction between PVT1 and EZH2. ChIP was used to study the promoter region of related genes. RESULTS: The higher expression of PVT1 was significantly correlated with deeper invasion depth and advanced TNM stage. Multivariate analyses revealed that PVT1 expression served as an independent predictor for overall survival (p = 0.031). Further experiments demonstrated that PVT1 knockdown significantly inhibited the proliferation both in vitro and in vivo. Importantly, we also showed that PVT1 played a key role in G1 arrest. Moreover, we further confirmed that PVT1 was associated with enhancer of zeste homolog 2 (EZH2) and that this association was required for the repression of p15 and p16. To our knowledge, this is the first report showed that the role and the mechanism of PVT1 in the progression of gastric cancer. CONCLUSIONS: Together, these results suggest that lncRNA PVT1 may serve as a candidate prognostic biomarker and target for new therapies in human gastric cancer.
Assuntos
Proliferação de Células/genética , Inibidor de Quinase Dependente de Ciclina p15/genética , Inibidor p16 de Quinase Dependente de Ciclina/genética , RNA Longo não Codificante/genética , Neoplasias Gástricas/genética , Animais , Apoptose/genética , Biomarcadores Tumorais/genética , Carcinogênese/genética , Linhagem Celular , Linhagem Celular Tumoral , Proteína Potenciadora do Homólogo 2 de Zeste , Epigenômica/métodos , Pontos de Checagem da Fase G1 do Ciclo Celular/genética , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Complexo Repressor Polycomb 2/genética , Prognóstico , Regiões Promotoras Genéticas/genética , Transfecção/métodosRESUMO
BACKGROUND: Increasing evidence indicates that long noncoding RNAs (IncRNAs) perform specific biological functions in diverse processes. Recent studies have reported that IncRNAs may be involved in ß cell function. The aim of this study was to characterize the role of IncRNA TUG1 in mouse pancreatic ß cell functioning both in vitro and in vivo. METHODS: qRT-PCR analyses were performed to detect the expression of lncRNA TUG1 in different tissues. RNAi, MTT, TUNEL and Annexin V-FITC assays and western blot, GSIS, ELISA and immunochemistry analyses were performed to detect the effect of lncRNA TUG1 on cell apoptosis and insulin secretion in vitro and in vivo. RESULTS: lncRNA TUG1 was highly expressed in pancreatic tissue compared with other organ tissues, and expression was dynamically regulated by glucose in Nit-1 cells. Knockdown of lncRNA TUG1 expression resulted in an increased apoptosis ratio and decreased insulin secretion in ß cells both in vitro and in vivo . Immunochemistry analyses suggested decreased relative islet area after treatment with lncRNA TUG1 siRNA. CONCLUSION: Downregulation of lncRNA TUG1 expression affected apoptosis and insulin secretion in pancreatic ß cells in vitro and in vivo. lncRNA TUG1 may represent a factor that regulates the function of pancreatic ß cells.
Assuntos
Células Secretoras de Insulina/metabolismo , Insulina/metabolismo , RNA Longo não Codificante/metabolismo , Animais , Apoptose , Proliferação de Células , Células Cultivadas , Regulação para Baixo , Glucose/farmacologia , Secreção de Insulina , Células Secretoras de Insulina/citologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos NOD , Pâncreas/metabolismo , Interferência de RNA , RNA Longo não Codificante/antagonistas & inibidores , RNA Longo não Codificante/genética , RNA Interferente Pequeno/metabolismoRESUMO
Colorectal cancer (CRC) remains an important public health problem in the world. Long noncoding RNA (lncRNA) is an RNA molecular that is longer than 200 nucleotides and cannot be translated into a protein. Recent studies have shown that lncRNAs play important roles in carcinogenesis and cancer metastasis. The aim of this study was to evaluate the expression and biological role of lncRNA maternally expressed gene 3 (MEG3) in colorectal cancer. Quantitative real-time-PCR (qRT-PCR) was performed to investigate the expression of MEG3 in tumor tissues and corresponding nontumor colorectal tissues from 62 patients. The lower expression of MEG3 was remarkably correlated with low histological grade, deep tumor invasion, and advanced tumor node metastasis (TNM) stage. Multivariate analyses revealed that MEG3 expression served as an independent predictor for overall survival. Further experiments revealed that overexpressed MEG3 significantly inhibited CRC cell proliferation both in vitro and in vivo. In conclusion, our study demonstrated that MEG3 is involved in the development and progression of colorectal cancer by regulating cell proliferation and shows that MEG3 may be a potential diagnostic and prognostic target in patients with colorectal cancer.
Assuntos
Biomarcadores Tumorais/biossíntese , Proliferação de Células/genética , Neoplasias Colorretais/genética , RNA Longo não Codificante/biossíntese , Idoso , Apoptose/genética , Biomarcadores Tumorais/genética , Linhagem Celular Tumoral , Neoplasias Colorretais/patologia , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Estimativa de Kaplan-Meier , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Prognóstico , RNA Longo não Codificante/genéticaRESUMO
BACKGROUND: Accumulating evidence indicates that the long non-coding RNA HOTAIR plays a critical role in cancer progression and metastasis. However, the overall biological role and clinical significance of HOTAIR in gastric carcinogenesis remains largely unknown. METHODS: HOTAIR expression was measured in 78 paired cancerous and noncancerous tissue samples by real-time PCR. The effects of HOTAIR on gastric cancer cells were studied by overexpression and RNA interference approaches in vitro and in vivo. Insights of the mechanism of competitive endogenous RNAs (ceRNAs) were gained from bioinformatic analysis, luciferase assays and RNA binding protein immunoprecipitation (RIP). The positive HOTAIR/HER2 interaction was identified and verified by immunohistochemistry assay and bivariate correlation analysis. RESULTS: HOTAIR upregulation was associated with larger tumor size, advanced pathological stage and extensive metastasis, and also correlated with shorter overall survival of gastric cancer patients. Furthermore, HOTAIR overexpression promoted the proliferation, migration and invasion of gastric carcinoma cells, while HOTAIR depletion inhibited both cell invasion and cell viability, and induced growth arrest in vitro and in vivo. In particular, HOTAIR may act as a ceRNA, effectively becoming a sink for miR-331-3p, thereby modulating the derepression of HER2 and imposing an additional level of post-transcriptional regulation. Finally, the positive HOTAIR/HER2 correlation was significantly associated with advanced gastric cancers. CONCLUSIONS: HOTAIR overexpression represents a biomarker of poor prognosis in gastric cancer, and may confer malignant phenotype to tumor cells. The ceRNA regulatory network involving HOTAIR and the positive interaction between HOTAIR and HER2 may contribute to a better understanding of gastric cancer pathogenesis and facilitate the development of lncRNA-directed diagnostics and therapeutics against this disease.
Assuntos
Biomarcadores Tumorais/genética , Carcinoma/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , RNA Longo não Codificante/genética , Receptor ErbB-2/genética , Neoplasias Gástricas/genética , Animais , Sequência de Bases , Biomarcadores Tumorais/metabolismo , Carcinoma/metabolismo , Carcinoma/mortalidade , Carcinoma/patologia , Proliferação de Células , Sobrevivência Celular , Feminino , Humanos , Metástase Linfática , Masculino , Camundongos , Camundongos Nus , MicroRNAs/metabolismo , Dados de Sequência Molecular , Transplante de Neoplasias , RNA Longo não Codificante/metabolismo , Receptor ErbB-2/metabolismo , Transdução de Sinais , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/mortalidade , Neoplasias Gástricas/patologia , Análise de Sobrevida , Microambiente TumoralRESUMO
In this paper, a novel three-dimensional (3D) porous lanthanide-organic framework, Eu2(µ4-pmdc)2(OH)2·3H2O (1), which is stable up to 400 °C, has been hydrothermally synthesized and characterized. It shows intriguing single-crystal-to-single-crystal transformation and reversible dehydration/rehydration phenomenon upon removal and rebinding of the lattice water molecules, which is supported by single-crystal X-ray diffraction, powder X-ray diffraction, and photoluminescence data.
Assuntos
Ácidos Carboxílicos/química , Európio/química , Pirimidinas/química , Água/química , Cristalização , Cristalografia por Raios X , Dessecação , Medições Luminescentes , Modelos Moleculares , Porosidade , Temperatura , Difração de Raios XRESUMO
Transfusion-related acute lung injury (TRALI) is a serious complication associated with blood transfusion and can cause transfusion associated fatalities. Both antibody dependent and non-dependent mechanisms are involved in TRALI, as proposed over the past years. Nonetheless, many details of the immune cells involved in TRALI, particularly the Mac1(+)/Gr1(+) cells from donors, are not fully understood yet. Here we used an in vitro transwell system and a mouse model to study the role of donor leukocytes, present in the donor material, in the occurrence of TRALI reactions. We found that there is a number of immature myeloid cells with Mac1(+)/Gr1(+) phenotype present in the red blood cell (RBC) products, when prepared by regular methods. We found that murine Mac1(+)/Gr1(+) cells from stored RBC products display an elevated MHC I and CD40 expression, as well as an enhanced tumor necrosis factor alpha(TNF-α), interlukin-6(IL-6) and macrophage inflammatory protein 2 (MIP-2) secretion. When tested in a transwell endothelial migration assay, Mac1(+)/Gr1(+) cells showed a significant capability to cross the endothelial barrier. In vivo investigation demonstrated that compared to the purified RBC transfusion, more murine Mac1(+)/Gr1(+) cells from the regular method produced RBC sequestered in the lung, which associated to shorter survival. Taken together, these data suggest that donor derived Mac1(+)/Gr1(+) cells can play a significant role in TRALI reactions, and that reduction of Mac1(+)/Gr1(+) cell number from RBC products is necessary to control the severity of TRALI reactions in clinic.
Assuntos
Lesão Pulmonar Aguda/etiologia , Células Mieloides/imunologia , Reação Transfusional , Lesão Pulmonar Aguda/imunologia , Adolescente , Adulto , Animais , Anticorpos/imunologia , Doadores de Sangue , Antígeno CD11b/biossíntese , Antígeno CD11b/imunologia , Citocinas/imunologia , Feminino , Citometria de Fluxo , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Pessoa de Meia-Idade , Modelos Animais , Receptores de Quimiocinas/biossíntese , Receptores de Quimiocinas/imunologia , Adulto JovemRESUMO
Product quality control is crucial for industrial production, but there is lack of simple and effective detect methods. In this study, the chromatographically pure N-hexane from different manufacturers and same manufacturers with different batches was detected with fluorescence fingerprint technology. The results showed that the fluorescence fingerprints of all samples were different from each other. The numbers of fluorescence peaks of the fingerprints of the famous international manufacturer was stable and the intensity was low. The chromatographically pure N-hexane made in China generally had more peaks, higher intensities and greater changes as compared to the imported product. This indicated that the domestic products had more impurities with high concentration and the product quality was unstable. The study showed that the fluorescence fingerprint can be used as a novel method for quality control of chemical reagents.
RESUMO
In recent years, three-dimensional fluorescence spectrometry has been widely used to study the transportation and transformation of the environment pollutants. But little understanding about the relationship between fluorescence characteristics and molecular structure restricts its application. In the present paper, the excitation-emission matrix (EEM) of the typical aromatic pollutants and isomers, phenanthrene and anthracene were studied. The result showed that there existed a peak locating at lambda ex/lambdaem = 225/340 nm in the EEM of both phenanthrene and anthracene. Furthermore, the peaks at 275/360 nm of phenanthrene located quite close to the peak of anthracene at 285/360 nm. However, the difference between the EEM of phenanthrene and anthracene was significant. There existed the third fluorescence peak at 275/340 nm and the most intensive peak at 225/340 nm in the EEM of phenanthrene. The EEM of anthracene was more complicated. The most intensive peaks located at lambda ex,/lambdaem = 250/ 380, 250/400 and 250/425 nm respectiveoy. In addition, the fluorescence intensity of anthracene at 225/340 nm was about 1. 63 times that of phenanthrene when their concentrations were about 0. 058 1 mg L-1. The orbital energy gap of the frontier molecules of phenanthrene and anthracene were 4. 779 and 3. 621 eV respectively according to the density functional theory. Owe to the smaller energy gap and better symmetry of electron cloud, anthracene was easier to be excited under the excitation of longer wavelength with higher fluorescence intensity. The density functional theory is a good tool to estimate the luminous capability of organic matters.
RESUMO
The present paper studied fluorescence fingerprint properties of the municipal wastewater with industrial wastewater as major components. There existed three typical fluorescence peaks in the excitation-emission matrix of the municipal wastewater, locating at about lambda(ex)/lambda(em) of 275/310, 230/340 and 220/310 nm respectively. The wastewater didn't display typical protein-like fluorescence as the municipal wastewater with domestic sewage as major component. The fluorescence intensity of the wastewater was quite high with remarkable difference between workday and weekend. These might relate to the high content of industrial wastewater. The advantages of the fluorescence fingerprint such as easy and fast measurement and rich information about the components of wastewater make it a novel tool in water quality monitoring and early-warning.
Assuntos
Resíduos Industriais/análise , Esgotos/análise , Águas Residuárias/análise , Água/análise , Monitoramento Ambiental , Fluorescência , Qualidade da ÁguaRESUMO
BACKGROUND: MicroRNAs (miRNAs) are short, non-coding RNAs (~22 nt) that play important roles in the pathogenesis of human diseases by negatively regulating gene expression. Although miR-196a has been implicated in several other cancers, its role in non-small cell lung cancer (NSCLC) is unknown. The aim of the present study was to examine the expression pattern of miR-196a in NSCLC and its clinical significance, as well as its biological role in tumor progression. METHODS: Expression of miR-196a was analyzed in 34 NSCLC tissues and five NSCLC cell lines by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The effect of DNA methylation on miR-196a expression was investigated by 5-aza-2-deoxy-cytidine treatment and bisulfite sequencing. The effect of miR-196a on proliferation was evaluated by MTT and colony formation assays, and cell migration and invasion were evaluated by transwell assays. Analysis of target protein expression was determined by western blotting. Luciferase reporter plasmids were constructed to confirm the action of miR-196a on downstream target genes, including HOXA5. Differences between the results were tested for significance using Student's t-test (two-tailed). RESULTS: miR-196a was highly expressed both in NSCLC samples and cell lines compared with their corresponding normal counterparts, and the expression of miR-196a may be affected by DNA demethylation. Higher expression of miR-196a in NSCLC tissues was associated with a higher clinical stage, and also correlated with NSCLC lymph-node metastasis. In vitro functional assays demonstrated that modulation of miR-196a expression affected NSCLC cell proliferation, migration and invasion. Our analysis showed that miR-196a suppressed the expression of HOXA5 both at the mRNA and protein levels, and luciferase assays confirmed that miR-196a directly bound to the 3'untranslated region of HOXA5. Knockdown of HOXA5 expression in A549 cells using RNAi was shown to promote NSCLC cell proliferation, migration and invasion. Finally, we observed an inverse correlation between HOXA5 and miR-196a expression in NSCLC tissues. CONCLUSIONS: Our findings indicate that miR-196a is significantly up-regulated in NSCLC tissues, and regulates NSCLC cell proliferation, migration and invasion, partially via the down-regulation of HOXA5. Thus, miR-196a may represent a potential therapeutic target for NSCLC intervention.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/patologia , Proteínas de Homeodomínio/metabolismo , Neoplasias Pulmonares/patologia , MicroRNAs/metabolismo , Apoptose/fisiologia , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Processos de Crescimento Celular/fisiologia , Linhagem Celular Tumoral , Movimento Celular/fisiologia , Metilação de DNA , Técnicas de Silenciamento de Genes , Proteínas de Homeodomínio/genética , Humanos , Neoplasias Pulmonares/metabolismo , MicroRNAs/biossíntese , MicroRNAs/genética , Invasividade Neoplásica , Regulação para CimaRESUMO
Macrophage activation is modulated by both environmental cues and endogenous programs. In the present study, we investigated the role of a PAQR family protein, monocyte to macrophage differentiation-associated (MMD), in macrophage activation and unveiled its underlying molecular mechanism. Our results showed that while MMD expression could be detected in all tissues examined, its expression level is significantly up-regulated upon monocyte differentiation. Within cells, EGFP-MMD fusion protein could be co-localized to endoplasmic reticulum, mitochondria, Golgi apparatus, but not lysosomes and cytoplasm. MMD expression is up-regulated in macrophages after LPS stimulation, and this might be modulated by RBP-J, the critical transcription factor of Notch signaling. Overexpression of MMD in macrophages increased the production of TNF-α and NO upon LPS stimulation. We found that MMD overexpression enhanced ERK1/2 and Akt phosphorylation in macrophages after LPS stimulation. Blocking Erk or Akt by pharmacological agent reduced TNF-α or NO production in MMD-overexpressing macrophages, respectively. These results suggested that MMD modulates TNF-α and NO production in macrophages, and this process might involves Erk or Akt.
Assuntos
MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Linfocinas/metabolismo , Macrófagos/enzimologia , Proteínas de Membrana/metabolismo , Óxido Nítrico/biossíntese , Proteínas Proto-Oncogênicas c-akt/metabolismo , Fator de Necrose Tumoral alfa/biossíntese , Animais , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Linhagem Celular , Ativação Enzimática/efeitos dos fármacos , Proteína de Ligação a Sequências Sinal de Recombinação J de Imunoglobina/metabolismo , Espaço Intracelular/efeitos dos fármacos , Espaço Intracelular/metabolismo , Lipopolissacarídeos/farmacologia , Linfocinas/genética , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Camundongos , Fosforilação/efeitos dos fármacos , Receptores Notch/metabolismo , Transdução de Sinais/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/genéticaRESUMO
Three-dimensional fluorescence spectroscopy is a new and effective chemical analysis method and is employed in water environment more and more widely. But the utilization is limited by the bottleneck, i. e. shortage of fluorescence data of contaminants in aqueous solution. This paper presents the three-dimensional fluorescence properties of a toxic contaminant, sodium butyl naphthalene sulfonate, in aqueous solution. There existed four peaks at about lambda(ex)/em = 230/340, 280/340, 225/650 and 280/650 nm respectively. The intensity of all the peaks except the peak at 225/650 nm increased as the concentration augmented, while the intensity of peak at 225/650 nm increased as the concentration augmented when the concentration was less than 0.5 mg x L(-1), and decreased as the concentration augmented when the concentration was greater than 0.5 mg x L(-1). The pH would lead to the variation in the fluorescence intensity vary rather than to change the peak location. The fluorescence intensities were stable when pH was in the range of 2-10. The study indicates that it is feasible to measure sodium butyl naphthalene sulfonate directly with the fluorescence intensity at 280/340 nm. The linear range is between 0 and 0.033 3 mg x L(-1). This simple and rapid method could provide reliable results without complex pretreatment.
RESUMO
BACKGROUND: Chronic HBV infection is a serious worldwide health problem that mainly causes liver cirrhosis and hepatocellular carcinoma (HCC). Few studies have explored how T cell exhaustion helps HBV avoid immune system attack and how to reverse that exhaustion. Recently, T cell immunoglobulin and immune receptor tyrosine-based inhibitory motif (ITIM) domain (TIGIT) have been identified as coinhibitory receptors, similar to PD-1. This study explores the expression of TIGIT and the T cell function changes in patients with chronic HBV infection. RESULTS: In this study, we found that the expression of TIGIT on T cells increased significantly in patients with chronic HBV infection. High expression of TIGIT on T cells is associated with functional exhaustion. Importantly, this study demonstrates that blocking TIGIT can reverse T cell exhaustion and restore function in patients with chronic HBV infection. CONCLUSIONS: HBV induces T cell exhaustion by up-regulating the expression of TIGIT. Blocking the TIGIT/PVR signaling pathway can reverse T cell exhaustion, so this discovery provides an immunotherapy target to battle chronic HBV infection.