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BACKGROUND: Ebosin is an exopolysaccharide produced by Streptomyces sp. 139, and its biosynthetic gene cluster (ste) has been previously described. Ste234 has high homology to the well-known ATP-binding cassette transport system DasABC, which has been linked to the regulation of morphological differentiation, antibiotics biosynthesis and aminosugars utilization in Streptomycetes. This study was conducted to evaluate the effect of the DasA family sugar binding protein Ste2 on Streptomyces sp. 139. RESULTS: The disruption of ste2 results in the upregulation of transcription of genes within Ebosin biosynthetic gene cluster and a two-fold increase in Ebosin production. RNA sequencing data suggests that the disruption of ste2 results in the decreased utilization of carbon and nitrogen sources, increased sensitivity to oxidative stress, as well as differed strain morphology, all of which have been experimentally proven. CONCLUSIONS: Taken together, Ste2 controls Ebosin yields, aminosugars uptake, sensitivity to oxidative stress, and morphological differentiation of Streptomyces sp. 139.
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Streptomyces , Família Multigênica , Nutrientes , Estresse Oxidativo , Streptomyces/genética , Streptomyces/metabolismo , Açúcares/metabolismoRESUMO
Histone deacetylases (HDACs) have been implicated in numerous biological events. However, to date, the role of HDAC6 in early embryos remains unknown. In the current study, Tubastatin A (TubA), a potent HDAC6 inhibitor, was used to block HDAC6 activity in mouse embryos. We found that TubA exposure significantly reduced the blastocyst formation of early embryos. Confocal microscopy revealed the markedly increased chromosomal congression failure in the mouse embryos treated with the HDAC6 inhibitor. Moreover, the HDAC6 inhibition resulted in the overproduction of reactive oxygen species (ROS) in embryos. In addition, we observed the accumulation of phosphorylated γH2AX in TubA-treated embryos, indicative of the increased DNA damage. In line with this, cell apoptosis of blastocysts was frequently detected in HDAC6-deficient embryos compared with their controls. Altogether, our data indicate that HDAC6 may serve as an important regulator of chromatin structure and mitochondrial function, determining the developmental potential of the early embryos of mouse.
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Desenvolvimento Embrionário/fisiologia , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Desacetilase 6 de Histona/metabolismo , Ácidos Hidroxâmicos/farmacologia , Indóis/farmacologia , Animais , Desenvolvimento Embrionário/efeitos dos fármacos , Feminino , Desacetilase 6 de Histona/antagonistas & inibidores , Desacetilase 6 de Histona/genética , Inibidores de Histona Desacetilases/farmacologia , Camundongos , Gravidez , Transporte Proteico , Espécies Reativas de OxigênioRESUMO
The clustered regularly interspaced short palindromic repeat (CRISPR)-Cas system has emerged as the dominating tool for genome engineering, while also changes the speed and efficiency of metabolic engineering in conventional and non-conventional yeasts. Among these CRISPR-Cas systems, CRISPR-Cas9 technology has usually been applied for removing unfavorable target genes. Here, we used CRISPR-Cas9 technology to delete the gal80 gene in uracil-deficient strain and had successfully remolded the engineered Saccharomyces cerevisiae that can produce artemisinic acid without galactose induction. An L9(34) orthogonal test was adopted to investigate the effects of different factors on artemisinic acid production. Fermentation medium III with sucrose as carbon sources, 1% inoculum level, and 84-h culture time were identified as the optimal fermentation conditions. Under this condition, the maximum artemisinic acid production by engineered S. cerevisiae 1211-2 was 740 mg/L in shake-flask cultivation level. This study provided an effective approach to reform metabolic pathway of artemisinic acid-producing strain. The engineered S. cerevisiae 1211-2 may be applied to artemisinic acid production by industrial fermentation in the future.
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Artemisininas/metabolismo , Galactose/metabolismo , Proteínas Repressoras/genética , Proteínas de Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Sistemas CRISPR-Cas , Fermentação , Deleção de Genes , Engenharia Metabólica , Proteínas Repressoras/metabolismo , Saccharomyces cerevisiae/crescimento & desenvolvimento , Proteínas de Saccharomyces cerevisiae/metabolismo , Uracila/metabolismoRESUMO
A PCR method is described to identify the species origin of various animal and human tissue-derived biochemical drugs. Four commercialized drugs, including spermary tablets, compound embryonic bovine liver extract tablets, spleen aminopeptide solution, and placenta polypeptide injection, were used as a proof-of-principle in this study. Primers were designed to amplify conservative regions of mitochondrial cytochrome b and ATPase 8 genes from beef, pork, lamb and human DNA, respectively. The specificity of primers for ATPase 8 gene is found to be higher than those for cytochrome b under the given experimental conditions. The amplicon sizes of ATPase 8 were 212, 271, 293 and 405 bp for pork, beef, lamb and human tissue, respectively. The minimum detectable concentration of DNA sample for species identification is 0.05-0.5 pg·µL-1. The species origin can be distinguished by this method in extremely low concentrations of template DNAs extracted. Conceivably, this PCR method for meat authentication may be extended to quality control of other biochemical drugs and raw materials. Graphical abstract A specific PCR method was developed for the detection of species origin in biochemical drugs via species-specific primers targeting mitochondrial ATPase 8 genes. The PCR products were separated by gel electrophoresis and species origins were indicated by comparison to references.
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Primers do DNA/genética , ATPases Mitocondriais Próton-Translocadoras/genética , Preparações Farmacêuticas/análise , Reação em Cadeia da Polimerase , Animais , Bovinos , Citocromos b/genética , DNA Mitocondrial/genética , Eletroforese em Gel de Ágar , Humanos , Carne/análise , Tamanho da Partícula , Ovinos , Propriedades de Superfície , Suínos , ComprimidosRESUMO
The objective of this study was to investigate the expression and significance of the transforming growth factor ß type II receptor (TGFßRII) in diffuse large B cell lymphoma. All patients were enrolled at the First Affiliated Hospital of Liaoning Medical University between 2001 and 2007. The median follow-up period was 53.3 months. Of the 338 patients studied, 131 (38.76 %) had TGFßRII positive expression on immunohistochemistry. The 5 year survival rate was significantly higher in patients with TGFßRII expression than in those without TGFßRII expression (40.3 vs. 31.6 %, P = 0.041). Multivariate analysis identified TGFßRII expression as an independent predictive parameter for survival, in addition to lactate dehydrogenase, clinical stage, and histologic subtype. TGFßRII expression may be considered a new prognostic factor of diffuse large B cell lymphoma.
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Biomarcadores Tumorais/genética , Linfoma Difuso de Grandes Células B/genética , Prognóstico , Proteínas Serina-Treonina Quinases/biossíntese , Receptores de Fatores de Crescimento Transformadores beta/biossíntese , Adulto , Idoso , Biomarcadores Tumorais/biossíntese , Feminino , Humanos , Imuno-Histoquímica , Linfoma Difuso de Grandes Células B/patologia , Masculino , Pessoa de Meia-Idade , Proteínas Serina-Treonina Quinases/genética , Receptor do Fator de Crescimento Transformador beta Tipo II , Receptores de Fatores de Crescimento Transformadores beta/genética , Transdução de Sinais , Análise de Sobrevida , Fator de Crescimento Transformador beta/genéticaRESUMO
Post-polycythemia vera myelofibrosis (post-PV MF) is a critical hematologic evolution of polycythemia vera (PV). The main purpose of the present study was to identify the possible risk factors for the occurrence and prognosis of post-PV MF in Chinese patients with PV. A cohort of 272 Chinese PV patients with JAK2(V617F) or exon12 mutation was retrospectively analyzed. Of the 272 patients with PV, 63 developed post-PV MF. Platelet count >550 × 10(9) /L and splenomegaly were identified as independent risk factors for post-PV MF. The median duration of survival for post-PV MF patients was 8 years. Anemia and age >65 years at diagnosis of post-PV MF were identified as significant predictors for the poor prognosis of post-PV MF. In conclusion, platelet counts and splenomegaly were significant predictors for the transformation to post-PV MF, while anemia (hemoglobin levels <100 g/L) and age>65 years were significant predictors for poor prognosis of post-PV MF in Chinese PV patients with JAK2(V617F) or exon12 mutation.
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Janus Quinase 2/genética , Policitemia Vera/complicações , Mielofibrose Primária/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Povo Asiático , Estudos de Coortes , Feminino , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Policitemia Vera/genética , Policitemia Vera/mortalidade , Prognóstico , Estudos Retrospectivos , Fatores de Risco , Análise de SobrevidaRESUMO
OBJECTIVE: To explore the survival and the risk factors of poor prognosis in Chinese patients with polycythemia vera (PV). METHODS: A total of 816 patients with a definite diagnosis of PV were enrolled from August 1983 to June 2013 into this study. The standardized mortality ratio (SMR) was calculated by comparing the cumulative survival of 816 PV patients with age- and sex- and calendar year-matched healthy Chinese population from the national bureau of statistics of the People's Republic of China. The clinical features of diagnosis and prognosis of PV patients were analyzed by Cox regression to identify risk factors for the poor prognosis of PV and to develop a dynamic prognostic model in Chinese patients. The effects of different treatments on the development of acute myelocytic leukemia (AML) and post-PV myelofibrosis (post-PV MF) were determined by Kaplan-Meier analysis. JAK2 V617F allele burden (V617F%) was determined by quantitative real-time PCR in 104 patients. RESULTS: The median follow-up time was 6 (1-42) years. The 10-, 15- and 20-year overall survival (OS) was 89.50%, 76.70% and 64.70%, respectively. The SMR was 17.40 (95% CI: 13.71-21.78). Cox regression analysis revealed that white blood cell (WBC) count>10×10(9)/L (HR=3.10, 95% CI: 1.47-6.53, P=0.003), age>60 years (HR=2.89, 95% CI: 1.84-4.53, P<0.001) and prior thrombosis (HR=2.66, 95% CI: 1.65-4.29, P<0.001) were significant predictors for the poor prognosis of PV. Based on the hazard radio, 816 patents were allocated into 4 categories with significantly different survival: low (sum of points=0; median survival no reached), intermediate 1 (sum of points=1; median survival 33.10 (28.20-38.00) years), intermediate 2 (sum of points=2; median survival 23.00 (16.08-29.92) years), high (sum of points=3; median survival 13.00 (10.58-15.42) years). The mortality of high risk group was 5.37 fold higher than low risk patients. The 10- and 20-year survival of no post-PV MF were 89.50% and 79.60%, respectively, for interferon α (IFN-α); 73.80% and 43.50%, respectively, for hydroxyurea treatment; 82.20% and 71.40%, respectively, for alkylating agent treatment; and 80.00% and 38.20%, respectively, for no cytoreductive treatment. The treatment of exposure to IFN-α associated with a higher rate of no-post-PV MF survival (Log-rank=9.79, P=0.020). There were more post-PV MF patients with V617F%≥50% compared with those V617F%<50% (P<0.001). CONCLUSIONS: The mortality of PV patients is significantly higher than that of healthy Chinese population. The WBC count>10×10(9)/L, age>60 years, and prior thrombosis are identified as significant predictors for the prognosis of PV. The risk of post-PV MF transformation may be ameliorated by IFN-α via decreasing the burden of JAK2 V617F mutation.
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Policitemia Vera , Mielofibrose Primária , Alelos , Povo Asiático , China , Humanos , Interferon-alfa , Estimativa de Kaplan-Meier , Leucemia Mieloide Aguda , Contagem de Leucócitos , Mutação , Prognóstico , Reação em Cadeia da Polimerase em Tempo Real , Análise de Regressão , Fatores de Risco , TromboseRESUMO
Chronic myelogenous leukemia (CML) is a complex disease with a genetic basis. The genetic association studies (GASs) that have investigated the association between adult CML and 5,10-methylenetetrahydrofolate reductase (MTHFR) C677T and A1298C polymorphisms have produced contradictory and inconclusive results. The aim of this meta-analysis is to provide a relatively comprehensive assessment of the association of these polymorphisms with adult CML risk. A literature search for eligible GAS published before September 15, 2013 was conducted in PubMed, Embase, Web of Science, Cochrane Library, and China National Knowledge Infrastructure (CNKI) databases. Pooled odds ratios (ORs) with their corresponding 95% confidence intervals (95% CIs) were used to evaluate the strength of the association under a fixed or random effect model according to heterogeneity test results. All analyses were performed using the Stata software, version 12.0. Twelve case-control studies were included in this meta-analysis with a total of 932 CML patients and 3,465 healthy controls. For MTHFR C677T (dbSNP: rs1801133, C>T), though the pooled ORs were not significant in the overall population, all the ORs greater than 1 suggested an increased risk of CML for carriers of the risk allele. However, stratified analysis based on genotyping method revealed a significant association in the PCR-restriction fragment length polymorphism (RFLP) subgroup, possibly as a result of heterogeneity. For MTHFR A1298C (dbSNP: rs1801131, A>C), the combined results showed that carriers of the C allele may be associated with a decreased risk of adult CML. Stratified analysis showed that the magnitude of this effect was especially significant among Asians, indicating ethnicity differences in adult CML susceptibility. This meta-analysis shows that the C allele of MTHFR A1298C may be associated with a decreased risk in adult CML, especially among Asians, while MTHFR C677T may not be associated with adult CML risk. However, the development of adult CML may be the result of gene-gene and gene-environment interactions, which should be considered in future individual GAS and subsequent meta-analyses.
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Predisposição Genética para Doença , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Polimorfismo Genético , Estudos de Associação Genética , Humanos , RiscoRESUMO
1. Nilotinib, a tyrosine kinase inhibitor, could potently inhibit SN-38 glucuronidation mainly catalyzed by UDP-glucuronosyltransferase (UGT) 1A1. This study was designed to investigate whether nilotinib can be used as a selective inhibitor of UGT1A1 in human liver. 2. Assays with recombinant UGTs indicated that nilotinib could strongly inhibit the activity of UGT1A1 and decreased the activity of extra-hepatic UGT1A7 to a much lesser extent. The inhibition on 4-methylumbelliferone (4Mu) glucuronidation by recombinant UGT1A1 obeyed competitive inhibition mechanism, with a kinetic constants (Ki) value of 0.17 µM. Assays with human liver microsomes (HLM) demonstrated that nilotinib could selectively inhibit estradiol-3-O-glucuronidation (E2-3-O-glucuronidation), a probe reaction of UGT1A1. Kinetic studies displayed that the inhibition on E2-3-O-glucuronidation followed non-competitive inhibition model, different from the inhibition on 4Mu glucuronidation. The Ki values were calculated to be 0.14 and 0.53 µM, depending on the enzyme sources of recombinant UGT1A1 or HLM, respectively. 3. Given that UGT1A7 is an extra-hepatic enzyme, this study indicates that nilotinib can be used as a selective inhibitor of UGT1A1 in human liver.
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Inibidores Enzimáticos/química , Glucuronosiltransferase/antagonistas & inibidores , Glucuronosiltransferase/metabolismo , Pirimidinas/química , Catálise , Glucuronídeos/metabolismo , Humanos , Cinética , Fígado/efeitos dos fármacos , Microssomos Hepáticos/metabolismo , Fenótipo , Proteínas Recombinantes/metabolismoRESUMO
PURPOSE: The objective of the study was to evaluate the use of tumor content in circulating cell-free DNA (ccfDNA) for monitoring hepatocellular carcinoma (HCC) throughout its natural history. EXPERIMENTAL DESIGN: We included 67 patients with hepatitis B virus-related HCC, of whom 17 had paired pre- and posttreatment samples, and 90 controls. Additionally, in a prospective cohort with hepatitis B virus surface antigen-positive participants recruited in 2012 and followed up biannually with blood sample collections until 2019, we included 270 repeated samples before diagnosis from 63 participants who later developed HCC (pre-HCC samples). Shallow whole-genome sequencing and the ichorCNA method were used to analyze genome-wide copy number and tumor content in ccfDNA. RESULTS: High tumor content was associated with advanced tumor stage (P < 0.001) and poor survival after HCC diagnosis [HR = 12.35; 95% confidence interval (CI) = 1.413-107.9; P = 0.023]. Tumor content turned negative after surgery (P = 0.027), whereas it remained positive after transarterial chemoembolization treatment (P = 0.578). In non-HCC samples, the mean tumor content (±SD) was 0.011 (±0.007) and had a specificity of 97.8% (95% CI = 92.2%-99.7%). In pre-HCC samples, the tumor content increased from 0.014 at 4 years before diagnosis to 0.026 at 1 year before diagnosis. The sensitivity of tumor content in detecting HCC increased from 22.7% (95% CI = 11.5%-37.8%) within 1 year before diagnosis to 30.4% (95% CI = 13.2%-52.9%) at the Barcelona Clinic Liver Cancer (BCLC) stage 0/A, 81.8% (95% CI = 59.7%-94.8%) at stage B, and 95.5% (95% CI = 77.2%-99.9%) at stage C. CONCLUSIONS: The tumor content in ccfDNA is correlated with tumor burden and may help in monitoring HCC 1 yearearlier than clinical diagnosis and in predicting patient prognosis.
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Biomarcadores Tumorais , Carcinoma Hepatocelular , Ácidos Nucleicos Livres , Neoplasias Hepáticas , Humanos , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/diagnóstico , Carcinoma Hepatocelular/sangue , Carcinoma Hepatocelular/patologia , Carcinoma Hepatocelular/terapia , Carcinoma Hepatocelular/mortalidade , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/diagnóstico , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/sangue , Neoplasias Hepáticas/terapia , Neoplasias Hepáticas/mortalidade , Masculino , Feminino , Pessoa de Meia-Idade , Biomarcadores Tumorais/genética , Ácidos Nucleicos Livres/sangue , Estudos Prospectivos , Idoso , Prognóstico , DNA Tumoral Circulante/genética , DNA Tumoral Circulante/sangue , Vírus da Hepatite B/isolamento & purificação , Vírus da Hepatite B/genética , Estadiamento de Neoplasias , AdultoRESUMO
Genome-wide circulating cell-free DNA (ccfDNA) fragmentation for cancer detection has been rarely evaluated using blood samples collected before cancer diagnosis. To evaluate ccfDNA fragmentation for detecting early hepatocellular carcinoma (HCC), we first modeled and tested using hospitalized HCC patients and then evaluated in a population-based study. A total of 427 samples were analyzed, including 270 samples collected prior to HCC diagnosis from a population-based study. Our model distinguished hospital HCC patients from controls excellently (area under curve 0.999). A high ccfDNA fragmentation score was highly associated with an advanced tumor stage and a shorter survival. In evaluation, the model showed increasing sensitivities in detecting HCC using 'pre-samples' collected ≥4 years (8.3%), 3-4 years (20.0%), 2-3 years (31.0%), 1-2 years (35.0%), and 0-1 year (36.4%) before diagnosis. These findings suggested ccfDNA fragmentation is sensitive in clinical HCC detection and might be helpful in screening early HCC.
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BACKGROUND: Accurate estimation of incidence and prevalence is vital for preventing and controlling diabetes. Administrative data (including insurance data) could be a good source to estimate the incidence of diabetes. However, how to determine the look-back period (LP) to remove cases with preceding records remains a problem for administrative data. A short LP will cause overestimation of incidence, whereas a long LP will limit the usefulness of a database. Therefore, it is necessary to determine the optimal LP length for identifying incident cases in administrative data. OBJECTIVE: This study aims to offer different methods to identify the optimal LP for diabetes by using medical insurance data from the Chinese population with reference to other diseases in the administrative data. METHODS: Data from the insurance database of the city of Weifang, China from between January 2016 and December 2020 were used. To identify the incident cases in 2020, we removed prevalent patients with preceding records of diabetes between 2016 and 2019 (ie, a 4-year LP). Using this 4-year LP as a reference, consistency examination indexes (CEIs), including positive predictive values, the κ coefficient, and overestimation rate, were calculated to determine the level of agreement between different LPs and an LP of 4 years (the longest LP). Moreover, we constructed a retrograde survival function, in which survival (ie, incident cases) means not having a preceding record at the given time and the survival time is the difference between the date of the last record in 2020 and the most recent previous record in the LP. Based on the survival outcome and survival time, we established the survival function and survival hazard function. When the survival probability, S(t), remains stable, and survival hazard converges to zero, we obtain the optimal LP. Combined with the results of these two methods, we determined the optimal LP for Chinese diabetes patients. RESULTS: The κ agreement was excellent (0.950), with a high positive predictive value (92.2%) and a low overestimation rate (8.4%) after a 2-year LP. As for the retrograde survival function, S(t) dropped rapidly during the first 1-year LP (from 1.00 to 0.11). At a 417-day LP, the hazard function reached approximately zero (ht=0.000459), S(t) remained at 0.10, and at 480 days, the frequency of S(t) did not increase. Combining the two methods, we found that the optimal LP is 2 years for Chinese diabetes patients. CONCLUSIONS: The retrograde survival method and CEIs both showed effectiveness. A 2-year LP should be considered when identifying incident cases of diabetes using insurance data in the Chinese population.
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Diabetes Mellitus , Seguro , Humanos , População do Leste Asiático , Estudos Retrospectivos , Diabetes Mellitus/epidemiologia , Povo AsiáticoRESUMO
CRISPR/Cas9 has been widely used in engineering Saccharomyces cerevisiae for gene insertion, replacement and deletion due to its simplicity and high efficiency. The selectable markers of CRISPR/Cas9 systems are particularly useful for genome editing and Cas9-plasmids removing in yeast. In our previous research, GAL80 gene has been deleted by the plasmid pML104-mediated CRISPR/Cas9 system in an engineered yeast, in order to eliminate the requirement of galactose supplementation for induction. The maximum artemisinic acid production by engineered S. cerevisiae 1211-2 (740 mg/L) was comparable to that of the parental strain 1211 without galactose induction. Unfortunately, S. cerevisiae 1211-2 was inefficient in the utilization of the carbon source ethanol in the subsequent 50 L pilot fermentation experiment. The artemisinic acid yield in the engineered S. cerevisiae 1211-2 was only 20%-25% compared with that of S. cerevisiae 1211. The mutation of the selection marker URA3 was supposed to affect the growth and artemisinic acid production. A ura3 mutant was successfully restored by a recombinant plasmid pML104-KanMx4-u along with a 90 bp donor DNA, resulting in S. cerevisiae 1211-3. This mutant could grow normally in a fed-batch fermentor with mixed glucose and ethanol feeding, and the final artemisinic acid yield (> 20 g/L) was comparable to that of the parental strain S. cerevisiae 1211. In this study, an engineered yeast strain producing artemisinic acid without galactose induction was obtained. More importantly, it was the first report showing that the auxotrophic marker URA3 significantly affected artemisinic acid production in a pilot-scale fermentation with ethanol feeding, which provides a reference for the production of other natural products in yeast chassis.
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Artemisininas , Proteínas de Saccharomyces cerevisiae , Fermentação , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismoRESUMO
Currently, multiple myeloma (MM) is still an incurable disease. Deciphering its pathogenesis will bring new targets for clinical diagnosis and treatment. In the present study, we identified a MM-associated circular RNA (circRNA), circ-MYBL2, which was dramatically decreased in MM tissue and serum samples in comparison to normal samples. Low circ-MYBL2 level was closely correlated with high clinical stage and unfavorable outcome, and serum circ-MYBL2 had excellent accuracy in diagnosing MM. Exogenous circ-MYBL2 expression notably repressed MM cell viability, DNA synthesis and cell cycle progression. Further exploration revealed that circ-MYBL2 exerted the tumor-inhibiting effect by affecting the phosphorylation level of its linear isoform, in which circ-MYBL2 facilitated the binding of Cyclin F to MYBL2, dampening MYBL2 phosphorylation and activation, thereby inhibiting the transcription of a number of well-known proliferation-related oncogenes. Importantly, overexpression of circ-MYBL2 significantly reduced the tumor size of subcutaneous xenografts in nude mice. Taken together, our data unveil a regulatory mechanism linking circ-MYBL2 and its host gene mediated by Cyclin F, providing a potential diagnostic, prognostic and therapeutic target for MM patients.
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Biomarcadores Tumorais/análise , Biomarcadores Tumorais/sangue , Proteínas de Ciclo Celular/análise , Proteínas de Ciclo Celular/sangue , Genes Supressores de Tumor , Mieloma Múltiplo/diagnóstico , Mieloma Múltiplo/genética , RNA Circular/análise , RNA Circular/sangue , Transativadores/análise , Transativadores/sangue , Animais , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Sobrevivência Celular/genética , Ciclinas , Expressão Gênica/genética , Xenoenxertos , Humanos , Camundongos Nus , Terapia de Alvo Molecular , Mieloma Múltiplo/patologia , Mieloma Múltiplo/terapia , Transplante de Neoplasias , Fosforilação , Transativadores/genética , Transativadores/metabolismoRESUMO
Members of the genus Streptomyces are known for their ability to produce compounds with various bioactivities and for their complex morphologies. Streptomyces sp. strain 139 is the producer strain of the exopolysaccharide (EPS) ebosin, which has remarkable in vivo antirheumatic arthritis activity. Here, we report its complete genome sequence, which will facilitate the study of the biosynthesis of ebosin.
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Rituximab, a CD20-reactive chimeric monoclonal antibody (mAb), induces apoptosis of lymphoma cells and promotes phagocytosis by dendritic cells and produces a cellular immunoresponse by cross-presentation of tumor antigens to T cells. Heat-stressed tumor cells also stimulate dendritic-cell maturation and induce specific cytotoxic T cells against tumor cells by inducing expression of heat-shock proteins. In this study, we used heat-stressed and rituximab-coated CD20+ lymphoma cells as antigens to load onto immature dendritic cells, and found that rituximab-coated CD20+ Raji cells could promote phagocytosis by dendritic cells, and that rituximab-coated or heat-stressed and then rituximab-coated CD20+ Raji cells resulted in increased dendritic cell maturation. Moreover, only CD20+ Raji cells treated with heat and rituximab effectively enhanced the immunostimulating functions of dendritic cells. Thus, our data indicate that rituximab and heat-shock proteins have synergistic effects, resulting in increased induction of cytotoxic T cells against B-cell lymphoma.
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Anticorpos Monoclonais/farmacologia , Antígenos CD20/imunologia , Antineoplásicos/farmacologia , Células Dendríticas/imunologia , Resposta ao Choque Térmico/imunologia , Linfoma/imunologia , Linfócitos T Citotóxicos/imunologia , Anticorpos Monoclonais Murinos , Linhagem Celular Tumoral , Proteínas de Choque Térmico/imunologia , Temperatura Alta , Humanos , Imunidade Celular/efeitos dos fármacos , Imunidade Celular/imunologia , RituximabRESUMO
Objective To explore whether Toll-like receptor 4 (TLR4) inhibits the proliferation and invasion of lymphoma cells through the nuclear factor-kappaB (NF-κB) signaling pathway and the down-regulation of Bcl2 and matrix metalloproteinase-9 (MMP-9). Methods Raji cells were treated with the TLR4 inhibitor TAK-242 at 10, 35, 60, 110, 170, 230 µmol/L for 2, 6, 24 hours. CCK-8 assay was used to select the optimal treatment concentration and time. The cells were then divided into control group and 10, 60 µmol/L TAK-242 (24-hour) groups. TranswellTM assay was performed to evaluate the cell invasion. Real-time quantitative PCR was used to detect the mRNA levels of TLR4 and NF-κBp65. Western blot analysis was used to test the protein levels of TLR4, NF-κBp65, Bcl2 and MMP-9. Results Compaired with the control group, the proliferation and invasion ablity of the Raji cells treated with TAK-242 (24 hour)apparently decreased. After the inhibition of TLR4, the mRNA and protein levels of NF-κBp65 were reduced, and the levels of Bcl2 and MMP-9 were also down-regualted. Conclusion Inhibition of TLR4 can decrease the proliferation and invation ability of lymphoma cells through the NF-κB pathway and the down-regulation of Bcl-2 and MMP-9 expression.
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Proliferação de Células , Linfoma/patologia , Transdução de Sinais , Receptor 4 Toll-Like/antagonistas & inibidores , Linhagem Celular Tumoral , Humanos , Metaloproteinase 9 da Matriz/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Fator de Transcrição RelA/metabolismoRESUMO
A molybdovanadosilicic acid H5SiMo11VO40·8H2O was synthesized and investigated in this work. The structure features and hydration degree of this acid were characterized by IR, UV, XRD and TG-DTA. Its proton conductivity was studied by electrochemical impedance spectroscopy (EIS). The EIS measurements demonstrated that H5SiMo11VO40·8H2O showed excellent proton conduction performance with proton conductivity reaching 5.70 × 10-3 S cm-1 at 26 °C and 70% relative humidity. So, it is a new solid high proton conductor. The conductivity enhances with the increase of temperature, and it exhibits Arrhenius behavior. The activation energy value for proton conduction is 21.4 kJ mol-1, suggesting that the proton transfer in this solid acid is dominated by Vehicle mechanism.
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Two vanadium-substituted polyoxometalate acid salt gel electrolytes, [PyPS]3H4SiW9V3O40 and [PyPS]5H2SiW9V3O40, have been synthesized using a 1-(3-sulfonic group) propylpyridine (PyPS) and a Keggin vanadium-substituted heteropoly acid H7SiW9V3O40 through an ionic self-assembly method, and adjusting the ratio of cation and anion. A substitution effect of the acid salt gel electrolytes has been investigated. Interestingly, when protons of the polyoxometalate acid salt gel electrolytes are substituted, both the conductivity and the phase transformation temperature increase. The fastest conductivity of these gel electrolytes was as high as 2.57 × 10-2 S cm-1 at 110 °C.
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Long non-coding RNAs (lncRNAs) are implicated in the complex network of cancer including Multiple myeloma (MM) and play important roles in tumor development. lncH19 was significantly up-regulated in multiple cancer types, suggesting it is a potential oncogene. However, the exact functions and downstream mechanisms are largely unknown. This study aimed to investigate whether H19 participates in the cell growth of MM and elucidate the underlying mechanism. We found that H19 was abnormally overexpressed in MM cell lines and sorted CD138+ MM bone marrow tissues. H19 knockdown induced by shRNA transfection significantly inhibited proliferation, viability and colony formation in MM cells, as well as inactivated NF-κB pathway. Moreover, combination treatment of H19 knockdown and NF-κB suppression (induced by specific inhibitor PDTC) produced synergistically inhibitory effects. Bone marrow expression of H19 was positively associated with circulating IL-6 or IL-8 level in the same MM patients. And patients with high expression of H19 had a lower survival rate. Taken together, we confirmed the abnormal upregulation of a novel lncRNA, H19, in human MM. H19 was involved in MM cell growth. The linkage between H19 and NF-κB pathway may provide a novel interpretation for the mechanism of H19's growth regulation in MM.