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BACKGROUND: Metazoan guts are in permanent contact with microbial communities. However, the host mechanisms that have developed to manage the dynamic changes of these microorganisms and maintain homeostasis remain largely unknown. RESULTS: Serotonin (5-hydroxytryptamine [5-HT]) was found to modulate gut microbiome homeostasis via regulation of a dual oxidase (Duox) gene expression in both Bactrocera dorsalis and Aedes aegypti. The knockdown of the peripheral 5-HT biosynthetic gene phenylalanine hydroxylase (TPH) increased the expression of Duox and the activity of reactive oxygen species, leading to a decrease in the gut microbiome load. Moreover, the TPH knockdown reduced the relative abundance of the bacterial genera Serratia and Providencia, including the opportunistic pathogens, S. marcescens and P. alcalifaciens in B. dorsalis. Treatment with 5-hydroxytryptophan, a precursor of 5-HT synthesis, fully rescued the TPH knockdown-induced phenotype. CONCLUSIONS: The findings reveal the important contribution of 5-HT in regulating gut homeostasis, providing new insights into gut-microbe interactions in metazoans.
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Microbioma Gastrointestinal , Animais , Microbioma Gastrointestinal/fisiologia , Homeostase , Insetos , Serotonina , SerratiaRESUMO
Population growth and industrial development have exacerbated environmental pollution of both land and aquatic environments with toxic and harmful materials. Luminescence-based chemical sensors crafted for specific hazardous substances operate on host-guest interactions, leading to the detection of target molecules down to the nanomolar range. Particularly, the luminescence-based sensors constructed on the basis of metal-organic frameworks (MOFs) are of increasing interest, as they can not only compensate for the shortcomings of traditional detection techniques, but also can provide more sensitive detection for analytes. Recent years have seen MOFs-based fluorescent sensors show outstanding advantages in the field of hazardous substance identification and detection. Here, we critically discuss the application of MOFs for the detection of a broad scope of hazardous substances, including hazardous gases, heavy metal ions, radioactive ions, antibiotics, pesticides, nitro-explosives, and some harmful solvents as well as luminous and sensing mechanisms of MOF-based fluorescent sensors. The outlook and several crucial issues of this area are also discussed, with the expectation that it may help arouse widespread attention on exploring fluorescent MOFs (LMOFs) in potential sensing applications.
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Estruturas Metalorgânicas , Metais Pesados , Corantes , Poluição Ambiental , Substâncias Perigosas , Íons , Estruturas Metalorgânicas/químicaRESUMO
Because H2O2 is thermally unstable, it seems to be difficult to synthesize peroxides at elevated temperatures. We describe here the in situ generation of peroxide that is incorporated in a new uranyl peroxo complex, HT-UPO1, through the hydrothermal treatment of uranyl nitrate at 150 °C in the presence of organic ligands. In this novel process, a highly conjugated aromatic carboxylate linker, (E)-4-[2-(pyridin-4-yl)vinyl]benzoic acid (HPyVB), plays a crucial role by inducing the reduction of oxygen in air to form peroxide in situ and coordinating with uranyl to promote the preferred formation of thermally stable HT-UPO1. This work expands our knowledge on the speciation and chemistry of uranyl peroxide compounds and also sheds light on the possibility of their synthesis under more harsh conditions.
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The separation of actinides has a vital place in nuclear fuel reprocessing, recovery of radionuclides, and remediation of environmental contamination. Here we propose a new paradigm of nanocluster-based actinide separation, namely, nanoextraction, that can achieve efficient sequestration of uranium in an unprecedented form of giant coordination nanocages using a cone-shaped macrocyclic pyrogallol[4]arene as the extractant. The U24-based hexameric pyrogallol[4]arene nanocages with distinctive [U2(PG)2] binuclear units (PG = pyrogallol) that rapidly assembled in situ in monophasic solvent were identified by single-crystal X-ray diffraction, MALDI-TOF mass spectrometry, NMR spectroscopy, and small-angle X-ray and neutron scattering. Comprehensive biphasic extraction studies showed that this novel separation strategy has enticing advantages such as fast kinetics, high efficiency, and good selectivity over lanthanides, thereby demonstrating its potential for efficient separation of actinide ions.
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The purpose of this study was to assess the value of metagenomic next-generation sequencing (mNGS) of bronchoalveolar lavage fluid (BALF) for the diagnosis of severe respiratory diseases based on interpretation of sequencing results. BALF samples were harvested and used for mNGS as well as microbiological detection. Infectious bacteria or fungi were defined according to relative abundance and number of unique reads. We performed mNGS on 35 BALF samples from 32 patients. The positive rate reached 100% in the mNGS analysis of nine immunocompromised patients. Compared with the culture method, mNGS had a diagnostic sensitivity of 88.89% and a specificity of 74.07% with an agreement rate of 77.78% between these two methods. Compared with the smear method and PCR, mNGS had a diagnostic sensitivity of 77.78% and a specificity of 70.00%. In 13 cases, detection results were positive by mNGS but negative by culture/smear and PCR. The mNGS findings in 11/32 (34.4%) cases led to changes in treatment strategies. Linear regression analysis showed that diversity was significantly correlated with interval between disease onset and sampling. Dynamic changes in reads could indirectly reflect therapeutic effectiveness. BALF mNGS improves sensitivity of pathogen detection and provides guidance in clinical practice. Potential pathogens can be identified based on relative abundance and number of unique reads.
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Infecções Bacterianas/diagnóstico , Líquido da Lavagem Broncoalveolar/microbiologia , Sequenciamento de Nucleotídeos em Larga Escala , Metagenômica/métodos , Micoses/diagnóstico , Adulto , Idoso , Bactérias/classificação , Bactérias/isolamento & purificação , Estado Terminal , Feminino , Fungos/classificação , Fungos/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Análise de Regressão , Sensibilidade e Especificidade , Manejo de EspécimesRESUMO
γδT cells are an important source of IL-17A and play an anti- or protumor role depending on the surrounding microenvironment. The precise role of γδT cells in the development of malignant pleural effusions (MPE) remains unknown. Using flow cytometry, we analyzed the distribution and differentiation of γδT cells in wild-type (WT) and IL-10-∕- mice. We carefully elucidated the influence of γδT cells on the formation of MPE by depleting γδT cells from WT, IL-10-∕-, and IL-17a-∕- mice. The distribution of γδT17 cells in human MPE and peripheral blood was also determined. Our data showed that both γδT cells and IL-17A-producing γδT (γδT17) cells accumulated in murine MPE, and IL-10 deficiency enhanced their accumulation. γδT cells were the main source of IL-17A in MPE for both WT and IL-10-∕- mice. IL-10 inhibited the chemotactic response of γδT cells to MPE and the proliferation of these cells. IL-10 suppressed γδT cell secretion of IL-17A via RORγt. The ablation of γδT cells accelerated MPE accumulation in both WT and IL-10-∕- mice, but it did not influence MPE accumulation in IL-17a-∕- mice. Patients with higher frequencies of γδT17 cells had significantly longer survival times than patients with lower frequencies of γδT17 cells. Taken together, our data demonstrate that γδT17 cells play an inhibitory role in the progression of MPE, and the accumulation of γδT17 cells in MPE is suppressed by IL-10.
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Interleucina-17/metabolismo , Linfócitos Intraepiteliais/citologia , Derrame Pleural Maligno/metabolismo , Animais , Diferenciação Celular , Separação Celular , Sobrevivência Celular , Quimiotaxia , Citometria de Fluxo , Humanos , Interleucina-10/metabolismo , Leucócitos Mononucleares/citologia , Neoplasias Pulmonares/sangue , Neoplasias Pulmonares/metabolismo , Complexo Principal de Histocompatibilidade , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microscopia de Fluorescência , Derrame Pleural Maligno/patologia , Células Th17/patologia , Microambiente TumoralRESUMO
Mutation in the gene encoding microphthalmia-associated transcription factor (MITF) lead to Waardenburg syndrome 2 (WS2), an autosomal dominantly inherited syndrome with auditory-pigmentary abnormalities, which is clinically and genetically heterogeneous. Haploinsufficiency may be the underlying mechanism for WS2. However, the mechanisms explaining the genotypic and phenotypic variations in WS2 caused by MITF mutations are unclear. A previous study revealed that MITF interacts with LEF-1, an important factor in the Wnt signaling pathway, to regulate its own transcription through LEF-1-binding sites on the MITF promoter. In this study, four different WS2-associated MITF mutations (p.R217I, p.R217G, p.R255X, p.R217del) that are associated with highly variable clinical features were chosen. According to the results, LEF-1 can activate the expression of MITF on its own, but MITF proteins inhibited the activation. This inhibition weakens when the dosage of MITF is reduced. Except for p.R217I, p.R255X, p.R217G, and p.R217del lose the ability to activate TYR completely and do not inhibit the LEF-1-mediated activation of the MITF-M promoter, and the haploinsufficiency created by mutant MITF can be overcome; correspondingly, the mutants' associated phenotypes are less severe than that of p.R217I. The dominant negative of p.R217del made it have a second-most severe phenotype. This study's data imply that MITF has a negative feedback loop of regulation to stabilize MITF gene dosage that involves the Wnt signaling pathway and that the interaction of MITF mutants with this pathway drives the genotypic and phenotypic differences observed in Waardenburg syndrome type 2 associated with MITF mutations.
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Genótipo , Fator de Transcrição Associado à Microftalmia/genética , Mutação , Fenótipo , Síndrome de Waardenburg/genética , Síndrome de Waardenburg/metabolismo , Via de Sinalização Wnt , Linhagem Celular , Epistasia Genética , Genes Reporter , Estudos de Associação Genética , Humanos , Fator 1 de Ligação ao Facilitador Linfoide/metabolismo , Modelos Biológicos , Regiões Promotoras Genéticas , Ligação ProteicaRESUMO
Waardenburg syndrome (WS) is an autosomal dominant inherited non-syndromic type of hereditary hearing loss characterized by varying combinations of sensorineural hearing loss and abnormal pigmentation of the hair, skin, and inner ear. WS is classified into four subtypes (WS1-WS4) based on additional symptoms. WS2 is characterized by the absence of additional symptoms. Recently, we identified a SOX10 missense mutation c.422T > C (p.L141P) associated with WS2. We performed functional assays and found the mutant loses DNA-binding capacity, shows aberrant cytoplasmic and nuclear localization, and fails to interact with PAX3. Therefore, the mutant cannot transactivate the MITF promoter effectively, inhibiting melanin synthesis and leading to WS2. Our study confirmed haploinsufficiency as the underlying pathogenesis for WS2.
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Haplótipos/genética , Fator de Transcrição PAX3/genética , Polimorfismo de Nucleotídeo Único/genética , Fatores de Transcrição SOXE/genética , Síndrome de Waardenburg/genética , Adolescente , Humanos , Masculino , Mutação/genéticaRESUMO
BACKGROUND: The role of combinations of tumor markers such as carcinoembryonic antigen (CEA), carbohydrate antigens (CA) 125, 15-3, and 19-9, and CYFRA 21-1 (a fragment of cytokeratin 19) in diagnosing malignant pleural effusion (MPE) has not been clearly established. OBJECTIVES: This meta-analysis was performed to establish the overall diagnostic accuracies of combinations of these pleural fluid tumor markers for MPE. METHODS: The PubMed, Ovid, Embase, Web of Science, and Cochrane bibliographic databases were searched. Sensitivity, specificity, and other measures of the accuracy of combinations of pleural CEA, CA 125, CA 15-3, CA 19-9, and CYFRA 21-1 in the diagnosis of MPE were pooled after a systematic review of English-language studies. RESULTS: Twenty studies met the inclusion criteria. For pleural fluid tumor marker combinations including more than 3 studies, the summary estimates of the sensitivity/specificity for diagnosing MPE were as follows: CEA + CA 125, 0.65/0.98; CEA + CA 15-3, 0.64/0.98; CEA + CA 19-9, 0.58/0.98; CEA + CYFRA 21-1, 0.82/0.92; and CA 15-3 + CYFRA 21-1, 0.88/0.94. CONCLUSIONS: In patients with undiagnosed pleural effusion, the combinations of positive pleural CEA + CA 15-3 and CEA + CA 19-9 are highly suspicious for pleural malignancy, but the sensitivity of these tests is poor. Therefore, their routine role in the diagnostic algorithm of these patients is questionable, and management decisions should depend on positive cytological or biopsy results from the pleura.
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Antígenos de Neoplasias/metabolismo , Antígeno Ca-125/metabolismo , Antígeno CA-19-9/metabolismo , Antígeno Carcinoembrionário/metabolismo , Queratina-19/metabolismo , Mucina-1/metabolismo , Derrame Pleural Maligno/metabolismo , Biomarcadores Tumorais/metabolismo , Humanos , Derrame Pleural Maligno/diagnóstico , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Family members provide essential support for ICU patients, contributing to their mental and physical recovery. Empowering ICU patients' families may help them overcome inadequacies and meet their own and patients' acknowledged needs. Nursing should understand and address patients' families' empowerment status. AIMS AND OBJECTIVES: To develop a tool, the Nurses' Empowerment Scale for Intensive Care Unit (ICU) Patients' Families (NESIPF), to help ICU nursing staff assess the empowerment status of patients' families. DESIGN: Four-phase instrument development study. METHODS: A 19-item instrument was initially generated based on literature review and interviews with family members of ICU patients. The Delphi research method was applied to gain expert opinion and consensus via rounds of questionnaires. A panel of 27 experts experienced in critical care medicine, nursing and psychology participated in two Delphi rounds and their input helped formulate an 18-item pretest instrument. Families of 20 patients were recruited to examine instrument readability. After a 2-week interval, another 20 patients' families were recruited to examine test-retest reliability. Two hundred questionnaires were then administered and analysed to examine the instrument's construct validity, criterion-related validity and internal consistency. RESULTS: Expert authority coefficients of two Delphi rounds reached 0·89 and 0·91. Kendall' W coefficients of 0·113 (P < 0·001) in round 1 and 0·220 (P < 0·001) in round 2 indicated slight to fair agreement among experts. Content validity index (CVI) reached 1·0 for 12 items; the CVI for item 13 was <0·7 so it was excluded. Cronbach's α coefficient was 0·92, indicating acceptable internal consistency reliability. The coefficient of internal consistency of each dimension was 0·717-0·921. The Pearson correlation coefficient >0·9 (P < 0·05) showed an acceptable test-retest reliability. CONCLUSIONS: The instrument has acceptable reliability and validity and can assess the empowerment status of families of critically ill patients. RELEVANCE TO CLINICAL PRACTICE: Knowledge of families' empowerment status may help to address their psychological needs and their ability to provide family support.
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Família/psicologia , Necessidades e Demandas de Serviços de Saúde , Recursos Humanos de Enfermagem Hospitalar/psicologia , Poder Psicológico , Inquéritos e Questionários , Adulto , Cuidados Críticos/psicologia , Técnica Delphi , Feminino , Humanos , Masculino , Pesquisa em Avaliação de Enfermagem , Psicometria , Reprodutibilidade dos TestesRESUMO
OBJECTIVE: Gastrointestinal stromal tumors (GISTs) have a broad spectrum of biological behaviors ranging from benign, borderline and malignant. This study aimed to screen differentially expressed microRNAs (miRNAs) between malignant and borderline GISTs and to investigate the potential role of miRNAs in the malignant transformation of GISTs. METHODS: Six GIST samples including borderline tumors (n = 3) and malignant tumors (n = 3) were collected based on the clinical and pathological characteristics. Total RNA was extracted, followed by miRNA microarray analysis to screen the differentially expressed miRNAs. The most significantly expressed 4 miRNAs were then chosen for further validation by real-time PCR in 22 additional GIST samples. RESULTS: Direct comparison of malignant group versus borderline group revealed 14 significantly and differentially expressed miRNAs (P < 0.05, with a fold change of < 0.5 or > 2). Five miRNAs were up-regulated and nine were down-regulated in the malignant group. Four miRNAs (miR-221, miR-135b, miR-675(*) and miR-218) were most significantly and differentially expressed between the two groups. The differential expression of 2 miRNAs (miR-221 and miR-675(*)) were subsequently confirmed with good concordance by real-time PCR. CONCLUSIONS: The differential miRNA expression profiles between two groups are revealed by miRNA microarray assay, and confirmed by real-time PCR. Among differentially expressed miRNAs, miR-221 and miR-675(*) might be related to the malignant transformation of GISTs, and have a potential value in predicting biological behavior of GISTs.
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Transformação Celular Neoplásica , Neoplasias Gastrointestinais/genética , Tumores do Estroma Gastrointestinal/genética , Perfilação da Expressão Gênica , MicroRNAs/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Regulação para Baixo , Feminino , Neoplasias Gastrointestinais/patologia , Tumores do Estroma Gastrointestinal/patologia , Humanos , Masculino , MicroRNAs/genética , Análise em Microsséries , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase em Tempo Real , Regulação para CimaRESUMO
The present paper is dedicated to the quantitative determination of oxygen-containing impurities in the LiF-NaF-KF eutectic using electrochemical (cyclic and square-wave voltammetry) and reduction melting methods. The LiF-NaF-KF melt was analyzed before and after purifying electrolysis. The amount of oxygen-containing impurities removed from the salt during purification was determined. It was found that after electrolysis, the concentration of oxygen-containing impurities decreased by 7 times. The results obtained via electrochemical techniques and reduction melting were well-correlated, which made it possible to evaluate the quality of the LiF-NaF-KF F melt. To verify the analysis conditions, mechanical mixtures of LiF-NaF-KF containing Li2O were analyzed using the reduction melting method. The oxygen concentration in the mixtures varied from 0.672 to 2.554 wt. %. Based on the analysis results, the dependence approximated by the straight line was obtained. These data may be used to draw calibration curves and to further develop the procedure of oxygen analysis of fluoride melts.
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BACKGROUND: Lung cancer is the second most commonly diagnosed cancer and the leading cause of cancer-related death. Malignant pleural effusion (MPE) is a special microenvironment for lung cancer metastasis. Alternative splicing, which is regulated by splicing factors, affects the expression of most genes and influences carcinogenesis and metastasis. METHODS: mRNA-seq data and alternative splicing events in lung adenocarcinoma (LUAD) were obtained from The Cancer Genome Atlas (TCGA). A risk model was generated by Cox regression analyses and LASSO regression. Cell isolation and flow cytometry were used to identify B cells. RESULTS: We systematically analyzed the splicing factors, alternative splicing events, clinical characteristics, and immunologic features of LUAD in the TCGA cohort. A risk signature based on 23 alternative splicing events was established and identified as an independent prognosis factor in LUAD. Among all patients, the risk signature showed a better prognostic value in metastatic patients. By single-sample gene set enrichment analysis, we found that among tumor-infiltrating lymphocytes, B cells were most significantly correlated to the risk score. Furthermore, we investigated the classification and function of B cells in MPE, a metastatic microenvironment of LUAD, and found that regulatory B cells might participate in the regulation of the immune microenvironment of MPE through antigen presentation and promotion of regulatory T cell differentiation. CONCLUSIONS: We evaluated the prognostic value of alternative splicing events in LUAD and metastatic LUAD. We found that regulatory B cells had the function of antigen presentation, inhibited naïve T cells from differentiating into Th1 cells, and promoted Treg differentiation in LUAD patients with MPE.
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Adenocarcinoma de Pulmão , Linfócitos B Reguladores , Neoplasias Pulmonares , Segunda Neoplasia Primária , Derrame Pleural Maligno , Humanos , Processamento Alternativo , Adenocarcinoma de Pulmão/genética , Prognóstico , Neoplasias Pulmonares/genética , Microambiente Tumoral/genéticaRESUMO
OBJECTIVE: To explore the clinical significance of KRAS mutation detection in colorectal adenocarcinoma. METHODS: Paraffin-embedded tissue specimens were obtained from 440 patients with colorectal adenocarcinoma. The genomic DNA was extracted. Mutations of exon 2 of KRAS gene were examined by PCR and direct sequencing. RESULTS: Somatic mutations of KRAS gene were identified in 146 cases, with the mutation rate of 33.2% (146/440). Among these 146 patients, KRAS mutation involved codon 12 in 118 patients, including 35G > A (Gly12Asp, 62 cases), 35G > T (Gly12Val, 35 cases), 34G > T (Gly12Cys, 9 cases), 34G > A (Gly12Ser, 6 cases), 35G > C (Gly12Ala, 5 cases), and 34G > C (Gly12Arg, 1 case); in 27 patients the mutation involved codon 13, including 38G > A (Gly13Asp, 25 cases), 38G > C (Gly13 Val, 1 case) and 37G > T (Gly13 Cys, 1 case); and in one patient, the mutation involved codon 14 with 40G > A (Val14Ile). The status of KRAS or codon 12 mutations in colorectal adenocarcinoma was related to patients' gender (P = 0.021 and P = 0.030, respectively), and this significant correlation to females was conserved in clinical stage III (P = 0.007 and P = 0.003, respectively), but not in stages I, II, and IV. The status of KRAS or codon 12 mutations was also related to tumor stage. Between stage II and stage IV, the mutation rate of KRAS and codon 12 showed significant difference (P = 0.028 and 0.034, respectively). Between stage III and stage IV, only the codon 12 mutation rate showed significant difference (P = 0.011). Codon 13 mutation was not related to tumor stage. CONCLUSION: About one third of patients with colorectal adenocarcinoma have KRAS gene mutation, which might be related to patients' gender; and could be consistently detected by PCR and direct sequencing.
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Adenocarcinoma/genética , Neoplasias Colorretais/genética , Mutação , Proteínas Proto-Oncogênicas/genética , Proteínas ras/genética , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Códon , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Éxons , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Reação em Cadeia da Polimerase , Proteínas Proto-Oncogênicas p21(ras) , Análise de Sequência de DNA , Fatores Sexuais , Adulto JovemRESUMO
OBJECTIVE: To investigate the clinical stage and histological grade of gastrointestinal stromal tumors. METHODS: Twelve clinical and pathological parameters were assessed in 613 patients with follow-up information. These parameters were classified into two gross spread parameters including liver metastasis and peritoneal dissemination, five microscopic spread parameters including lymph node metastasis, vascular, fat, nerve and mucosal infiltration, and five histological parameters including mitotic count ≥ 10 per 50 high-power fields, muscularis propria infiltration, coagulative necrosis, perivascular pattern and severe nuclear atypia. RESULTS: The accumulated 5-year disease-free survival (DFS) and overall survival (OS) of 293 patients without any of these predictive parameters of malignancy were 99.3% and 100.0%, respectively. They were regarded as nonmalignant and further evaluations on the stage and grade of these tumors were not performed. At least one and at most seven predictive parameters of malignancy were identified in 320 patients. For these patients, the accumulated 5-year DFS and OS rates were 43.9% (mean 6.7 years) and 59.7% (mean 9.3 years), respectively. The DFS showed significant difference between patients with and without gross spread (P < 0.01), with and without microscopic spread (P = 0.001). DFS and OS were associated with the number of predictive parameters of malignancy in patients without gross spread (P < 0.01 for both DFS and OS), but not in patients with gross spread (P = 0.882 and 0.441, respectively). CONCLUSIONS: Malignant GIST could be divided into clinical stages I and II based on the absence and presence of gross spread, respectively. The degree of malignancy of patients in clinical stage I could be graded according to the number of predictive parameters of malignancy. Patients in clinical stage II were of the highest degree of malignancy regardless of the number of parameters. The staging and grading of gastrointestinal stromal tumors in this study are strongly associated with prognosis.
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Tumores do Estroma Gastrointestinal/patologia , Gradação de Tumores/métodos , Estadiamento de Neoplasias/métodos , Actinas/metabolismo , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígenos CD34/metabolismo , Intervalo Livre de Doença , Feminino , Seguimentos , Tumores do Estroma Gastrointestinal/metabolismo , Tumores do Estroma Gastrointestinal/cirurgia , Humanos , Neoplasias Hepáticas/secundário , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica , Proteínas Proto-Oncogênicas c-kit/metabolismo , Taxa de Sobrevida , Adulto JovemRESUMO
The arylalkylamine N-acetyltransferase (AANAT) enzymes catalyze the acetyl-CoA-dependent acetylation of an amine or arylalkylamine, which is involved in important biological processes of insects. Here, we carried out the molecular and biochemical identiï¬cation of an arylalkylamine N-acetyltransferase (AANAT) from the oriental fruit fly, Bactrocera dorsalis. Using a bacterial expression system, we expressed and puriï¬ed the encoded recombinant BdorAANAT1-V3 protein. The puriï¬ed recombinant protein acts on a wide range of substrates, including dopamine, tyramine, octopamine, serotonin, methoxytryptamine, and tryptamine, and shows similar substrate afï¬nity (i.e., Km values: 0.16-0.26 mM) except for serotonin (Km = 0.74 mM) and dopamine (Km = 0.84 mM). Transcriptional proï¬le analysis of BdorAANAT1 revealed that this gene is most prevalent in adults and abundant in the adult brain, gut, and ovary. Using the CRISPR/Cas9 technique, we successfully obtained a BdorAANAT1 knockout strain based on a wild-type strain (WT). Compared with the WT, the cuticle color of larvae and pupae is normal; however, in adult mutants, the yellow region of their thorax is darkly pigmented, and two black spots were evident at the abdomen's end. Moreover, the female BdorAANAT1 knockout mutant had a smaller ovary than the WT, and laid far fewer eggs. Loss of function of BdorAANAT1 caused by RNAi with mature adult females in which the reproductive system is fully developed had no effect on their fecundity. Altogether, these results indicate that BdorAANAT1 regulates ovary development. Our findings provide evidence for the insect AANAT1 modulating adult cuticle pigmentation and female fecundity.
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Arilalquilamina N-Acetiltransferase , Tephritidae , Feminino , Animais , Arilalquilamina N-Acetiltransferase/química , Dopamina/metabolismo , Serotonina/metabolismo , Ovário/metabolismo , Tephritidae/genética , Tephritidae/metabolismo , Pigmentação/genética , Proteínas Recombinantes/genética , Drosophila/metabolismoRESUMO
Muscarinic acetylcholine receptors (mAChRs) play important roles in the insect nervous system. These receptors are G protein-coupled receptors, which are potential targets for insecticide development. While the investigation of pharmacological properties of insect mAChRs is growing, the physiological roles of the receptor subtype remain largely indeterminate. Here, we identiï¬ed three mAChR genes in an important agricultural pest Bactrocera dorsalis. Phylogenetic analysis deï¬ned these genes as mAChR-A, -B, and -C. Transcripts of the three mAChRs are most prevalent in 1-d-old larvae and are more abundant in the brain than other body parts in adults. Functional assay of Bdor-mAChR-B transiently expressed in Chinese hamster ovary cells showed that it was activated by acetylcholine (EC50, 205.11 nM) and the mAChR agonist oxotremorine M (EC50, 2.39 µM) in a dose-dependent manner. Using the CRISPR/Cas9 technique, we successfully obtained a Bdor-mAChR-B knockout strain based on wild-type (WT) strain. When compared with WT, the hatching and eclosion rate of Bdor-mAChR-B mutants are significantly lower. Moreover, the crawl speed of Bdor-mAChR-B knockout larvae was lower than that of WT, while climbing performance was enhanced in the mutant adults. Adults with loss of function of Bdor-mAChR-B showed declined copulation rates and egg numbers (by mated females). Our results indicate that Bdor-mAChR-B plays a key role in the development, locomotion, and mating behavior of B. dorsalis.
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Acetilcolina/farmacologia , Proteínas de Insetos/genética , Agonistas Muscarínicos/farmacologia , Oxotremorina/análogos & derivados , Receptores Muscarínicos/genética , Tephritidae/genética , Animais , Sequência de Bases , Proteínas de Insetos/metabolismo , Masculino , Oxotremorina/farmacologia , Filogenia , Receptores Muscarínicos/metabolismo , Alinhamento de Sequência , Tephritidae/metabolismoRESUMO
Electrorefining process has been widely used to separate and purify metals, but it is limited by deposition potential of the metal itself. Here we report in-situ anodic precipitation (IAP), a modified electrorefining process, to purify aluminium from contaminants that are more reactive. During IAP, the target metals that are more cathodic than aluminium are oxidized at the anode and forced to precipitate out in a low oxidation state. This strategy is fundamentally based on different solubilities of target metal chlorides in the NaAlCl4 molten salt rather than deposition potential of metals. The results suggest that IAP is able to efficiently and simply separate components of aluminum alloys with fast kinetics and high recovery yields, and it is also a valuable synthetic approach for metal chlorides in low oxidation states.
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OBJECTIVE: To study the clinicopathologic features, differential diagnosis and pathogenesis of sclerosing angiomatoid nodular transformation of spleen. METHODS: Ten cases of sclerosing angiomatoid nodular transformation of spleen were retrieved from the archival file. Histochemical and immunohistochemical (EnVision method) studies were performed. Ultrastructural findings were also available in one of them. RESULTS: Sclerosing angiomatoid nodular transformation was characterized by micronodular appearance of vascular spaces lined by plump endothelial cells with interspersed ovoid spindle cells. Immunohistochemical study showed that the endothelial cells of vessels in the angiomatoid nodules had various expressions of immunologic phenotypes and could be mainly classified into 3 types: CD34(+)/CD31(+)/CD8â» endothelial cells of the capillaries, CD8(+)/CD31(+)/CD34â» lining cells of the sinusoids and CD31(+)/CD8â»/CD34â» endothelial cells of the small veins. Collagen network and dilated lymphatic sinuses were evident under transmission electron microscope. CONCLUSIONS: Sclerosing angiomatoid nodular transformation of spleen is a rare benign entity. It may represent a reactive condition and bears some relationship with splenic angioma. It needs to be distinguished from borderline or malignant vascular tumors of spleen.
Assuntos
Histiocitoma Fibroso Benigno/patologia , Neoplasias Esplênicas/patologia , Adulto , Antígenos CD34/metabolismo , Antígenos CD8/metabolismo , Diagnóstico Diferencial , Feminino , Hemangioendotelioma/metabolismo , Hemangioendotelioma/patologia , Hemangiossarcoma/metabolismo , Hemangiossarcoma/patologia , Histiocitoma Fibroso Benigno/metabolismo , Histiocitoma Fibroso Benigno/cirurgia , Histiocitoma Fibroso Benigno/ultraestrutura , Humanos , Masculino , Microscopia Eletrônica , Pessoa de Meia-Idade , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Neoplasias Esplênicas/metabolismo , Neoplasias Esplênicas/cirurgia , Neoplasias Esplênicas/ultraestruturaRESUMO
OBJECTIVE: To investigate possible differences in the prognosis in children with severe nocturia who received different drug withdrawal schedules. METHODS: Ninety-seven children with severe nocturia were randomly assigned to two groups: control (n=47) and observed (n=50). The control group accepted drug withdrawal immediately, while the observed group accepted dose tapering gradually after a 12-week treatment course. The frequency of enuresis was observed three months after complete drug withdrawal. RESULTS: During the treatment, the frequency of enuresis in all of children from both the control and the observed groups was reduced by over 90%. Forty-six children (92%) from the observed group showed the frequency of enuresis was reduced by over 90%, but 28 children (60%) from the control group (p<0.01) three months after the complete drug withdrawal. There were no significant differences in the adverse effect and the medication compliance between the two groups. CONCLUSIONS: The different schedules of drug withdrawal may lead to different prognosis, and the schedule of gradual drug withdrawal may be superior to the immediate one in children with nocturnal enuresis.