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As one of the most widely used herbicides in agricultural industry, the residues of glyphosate (GLY) are frequent environmental pollutants. Freshwater planarian Dugesia japonica has been developed as a model for neurotoxicology. In this study, the effects of GLY on locomotion and feeding behavior, as well as neuroenzyme activities and mRNA expressions of D. japonica were determined. Additionally, histochemical localization was executed to explore the damage to the central nervous system (CNS) of planarians stressed by GLY. The results showed that the locomotor velocity, ingestion rate and the neuroenzyme activity were inhibited and the gene expressions were altered. Also, histo-architecture injury to CNS of planarians upon GLY exposure in a time-dependent manner was observed. Collectively, our results indicate that GLY can cause neurotoxicity to freshwater planarians representing as reduction in locomotor velocity and feeding rate by disturbing the neurotransmission systems and damaging the structure of CNS.
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Planárias , Animais , Planárias/genética , Glicina/toxicidade , Glicina/metabolismo , GlifosatoRESUMO
Glyphosate (GLY) is one of the most widely used agrochemicals in the world, and its exposure has become a public health concern. The freshwater planarian is an ideal test organism for detecting the toxicity of pollutants and has been an emerging animal model in toxicological studies. Nevertheless, the underlying toxicity mechanism of GLY to planarians has not been thoroughly explored. To elucidate the toxicity effects and molecular mechanism involved in GLY exposure of planarians, we studied the acute toxicity, histological change, and transcriptional response of Dugesia japonica subjected to GLY. Significant morphological malformations and histopathological changes were observed in planarians after GLY exposure for different times. Also, a number of differentially expressed genes (DEGs) were obtained at 1, 3 and 5 d after exposure; Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of these DEGs were performed, and a global and dynamic view was obtained in planarians upon GLY exposure at the transcriptomic level. Furthermore, real-time quantitative PCR (qRT-PCR) was conducted on nine DEGs associated with detoxification, apoptosis, stress response, DNA repair, etc. The expression patterns were well consistent with the RNA sequencing (RNA-seq) results at different time points, which confirmed the reliability and accuracy of the transcriptome data. Collectively, our results established that GLY could pose adverse effects on the morphology and histo-architecture of D. japonica, and the planarians are capable of responding to the disadvantageous stress by dysregulating the related genes and pathways concerning immune response, detoxification, energy metabolism, DNA damage repair, etc. To the best of our knowledge, this is the first report of transcriptomic analyses of freshwater planarians exposed to environmental pollutants, and it provided detailed sequencing data deriving from transcriptome profiling to deepen our understanding the molecular toxicity mechanism of GLY to planarians.
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Poluentes Ambientais , Herbicidas , Planárias , Poluentes Químicos da Água , Animais , Planárias/genética , Herbicidas/toxicidade , Herbicidas/análise , Reprodutibilidade dos Testes , Poluentes Químicos da Água/toxicidade , Poluentes Ambientais/farmacologia , GlifosatoRESUMO
OBJECTIVE: To investigate the diagnostic performance of the apparent diffusion coefficient (ADC) derived from intratumoral and peritumoral zones for assessing pathologic prognostic factors in rectal cancer. MATERIALS AND METHODS: One hundred forty-six patients with rectal cancer who underwent preoperative MRI were prospectively enrolled. Two radiologists independently placed free-hand regions of interest (ROIs) in the largest tumor cross section and three small ROIs on the peritumoral zone adjacent to the tumor contour. Maximum values of tumor ADC (ADCtmax), minimum values of tumor ADC (ADCtmin), mean values of tumor ADC (ADCtmean), mean values of peritumor ADC (ADCpmean), and ADCpmean/ADCtmean (ADC ratio) were obtained on ADC maps and correlated with prognostic factors using uni- and multivariate logistic regression, and receiver operating characteristic curve (ROC) analysis. RESULTS: Interobserver agreement was excellent for ADCtmax and ADCtmean (intraclass correlation coefficient [ICC], 0.915-0.958), and were good for ADCtmin, ADCpmean, and ADC ratio (ICC, 0.774-0.878). The ADC ratio was significantly higher in the poor differentiation, T3-4 stage, lymph node metastasis (LNM)-positive, extranodal extension (ENE)-positive, tumor deposit (TD)-positive, and lymphovascular invasion (LVI)-positive groups than that in the well-moderate differentiation, T1-2 stage, LNM-negative, ENE-negative, TD-negative, and LVI-negative groups (p = 0.008, < 0.001, < 0.001, 0.001, < 0.001, and < 0.001, respectively). The area under the ROC curve (AUC) of the ADC ratio was the highest for assessing poor differentiation (0.700), T3-4 stage (0.707), LNM-positive (0.776), TD-positive (0.848), and LVI-positive (0.778). Both the ADC ratio (AUC = 0.677) and ADCpmean (AUC = 0.686) showed higher diagnostic performance for assessing ENE. CONCLUSION: The ADC ratio could provide better predictive performance for assessing preoperative prognostic factors in resectable rectal cancer. KEY POINTS: ⢠Both the peritumor/tumor ADC ratio and ADCpmean are correlated with important prognostic factors of resectable rectal cancer. ⢠Both peritumor ADC and peritumor/tumor ADC ratio had higher diagnostic performance than tumor ADC for assessment of prognostic factors in resectable rectal cancer. ⢠Peritumor/tumor ADC ratio showed the most capability for the assessment of prognostic factors in resectable rectal cancer.
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Imagem de Difusão por Ressonância Magnética , Neoplasias Retais , Humanos , Metástase Linfática , Imageamento por Ressonância Magnética , Prognóstico , Neoplasias Retais/diagnóstico por imagem , Neoplasias Retais/patologia , Estudos RetrospectivosRESUMO
OBJECTIVE: This study investigates the association of the liver and spleen signal intensity on MRI with anemia in patients with gynecologic cancer. METHODS: 332 patients with gynecological cancer and 78 healthy women underwent MRI examination. Liver and spleen MRI parameters and laboratory tests were obtained within 1 week. The signal intensity ratios of liver and spleen to the paraspinal muscle were calculated on gradient-echo T1-weighted images (T1WI) and T2-weighted images (T2WI) in both patients and healthy women, respectively. RESULTS: The ratios of liver and spleen to paraspinal muscle on T1WI and T2WI were lower in patients than in the healthy women, respectively (P<0.0001). The ratios of the liver and spleen to paraspinal muscle on T1WI and T2WI decreased with the increasing stage of anemia and decreasing hemoglobin levels (P<0.001). The ratios of the liver to paraspinal muscle on T1WI, spleen to paraspinal muscle on T1WI, and the liver and spleen to paraspinal muscle on T2WI could predict anemia stage≥1 (AUC=0.576, 0.643, 0.688, and 0.756, respectively), ≥2 (AUC=0.743, 0.714, 0.891, and 0.922, respectively) and 3 (AUC=0.851, 0.822, 0.854, and 0.949, respectively). CONCLUSION: T2WI-based spleen signal intensity ratios showed the highest potential for non-invasive evaluation of anemia in gynecological cancer.
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Anemia , Neoplasias , Anemia/diagnóstico por imagem , Feminino , Humanos , Fígado/diagnóstico por imagem , Imageamento por Ressonância Magnética/métodos , Baço/diagnóstico por imagemRESUMO
OBJECTIVE: To assess the role of region of interest (ROI) selection of intravoxel incoherent motion (IVIM) for predicting lymph node metastases (LNM) and tumor response after chemoradiation therapy (CRT) in locally advanced rectal cancer. MATERIALS AND METHODS: Seventy-nine patients with biopsy-proven rectal adenocarcinoma who underwent pre- and post-CRT MRI and surgery were prospectively enrolled. The exclusion criteria included nonresectable and/or metastatic disease and loss of follow-up. Pathological stage was determined using ypTNM stage and tumor regression grade. Slow diffusion coefficient (D), fast diffusion coefficient (D*), perfusion-related diffusion fraction (f), apparent diffusion coefficient (ADC) and their percentage changes (Δ%) were evaluated by two readers using whole-volume, single-slice and small samples ROI methods. Risk factors including carcinoembryonic antigen, post-CRT T-staging, extramural venous invasion and IVIM parameters were evaluated through multivariate analyses. Areas under the receiver operating characteristic curves (AUCs) were calculated to evaluate diagnostic performance. Duration of follow-up was two-year. Recurrence-free survival of patients with LNM and tumor response was estimated using Kaplan-Meier analysis. RESULTS: Interobserver agreement were good for pre- and post-CRT three ROI methods (intraclass correlation coefficient [ICC], 0.581-0.953). Whole-volume ROI-derived Δ%D was an independent risk factor for LNM, non-pathological complete response (non-pCR) and poor response (odds ratio, 0.940, 0.952, 0.805, respectively; all p < 0.001). Whole-volume ROI-derived Δ%D showed best AUC of 0.810, 0.851 and 0.903 for LNM, non-pCR and poor response (cutoff value, 31.8%, 54.5%, 52.8%, respectively). Patients with post-CRT LNM showed reduction in 2-year recurrence-free survival (hazard ratio, 3.253). CONCLUSIONS: Whole-volume ROI-derived Δ%D provided high diagnostic performance for evaluating post-CRT LNM and tumor response. Patients with post-CRT LNM showed earlier recurrence.
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Quimiorradioterapia , Neoplasias Retais , Imagem de Difusão por Ressonância Magnética , Humanos , Linfonodos/diagnóstico por imagem , Movimento (Física) , Neoplasias Retais/tratamento farmacológico , Neoplasias Retais/terapia , RetoRESUMO
BACKGROUND: Freshwater planarians of the genus Dugesia (Platyhelminthes, Tricladida, Dugesiidae) are distributed in a major part of the Old World and Australia, although until recently only very few species were known from China. RESULTS: Two new species of Dugesia from Southern China are described on the basis of an integrative taxonomic approach. BI and ML phylogenetic trees based on the independent genes and on the concatenated dataset had similar topologies, only differing in some nodes that were weakly supported. Phylogenetic trees based on the concatenated dataset revealed that D. adunca Chen & Sluys, sp. nov. and D. tumida Chen & Sluys, sp. nov. are not closely related and belong to different clades. The two new species occupy separate long branches with high support values and, thus, are well-differentiated from their congeners. Separate species status of D. adunca and D. tumida is supported also by the genetic distances between the species included in our analysis, albeit that COI distances varied greatly among species. Dugesia adunca from Guangxi Province is characterized by the following features: living mature animals rather small; asymmetrical openings of the oviducts into the bursal canal; penis papilla with shape of an aquiline bill, albeit with a blunt tip; asymmetrical penis papilla, with a large antero-dorsal lip and a much smaller ventro-posterior lip; very large seminal vesicle, provided with trabeculae; small diaphragm; mixoploid karyotype with diploid complements of 2n = 2x = 16 and triploid complements of 2n = 3x = 24, with all chromosomes being metacentric. Dugesia tumida from Guangdong Province is characterized by a penis papilla provided with a large, symmetrical penial valve from the middle of which arises the small, distal section of the papilla; a duct intercalated between the seminal vesicle and the small diaphragm; ventrally displaced ejaculatory duct curving upwards before opening to the exterior; penis papilla highly asymmetrical, having a slim and long ventral portion and a short and stubby dorsal part; vasa deferentia separately opening into antero-dorsal portion of seminal vesicle; oviducts openings symmetrically into ventral portion of the bursal canal, near its opening into the atrium; mixoploid karyotype, with diploid chromosome portraits of 2n = 2x = 16, and triploid complements of 2n = 3x = 24, with all chromosomes being metacentric. In the context of the various kinds of mixoploidy and the sexualization of specimens, reproductive modalities within the genus Dugesia are shortly discussed. CONCLUSION: Molecular, morphological, and karyological markers show that the two populations examined represent members of the genus Dugesia and constitute two new, distinct species.
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Two new species of the genus Dugesia (Platyhelminthes, Tricladida, Dugesiidae) from the tropical monsoon forest in southern China are described on the basis of an integrative taxonomic study involving morphology, karyology, histology, and molecular analyses. The new species Dugesiacircumcisa Chen & Dong, sp. nov. is characterised by asymmetrical openings of the oviducts; right vas deferens opening at anterior portion of the seminal vesicle and the left one opening at mid-lateral portion of the seminal vesicle; two diaphragms in ejaculatory duct, the latter being ventrally displaced and opening at the tip of the penis papilla, which is provided with a nozzle; wide duct connecting male atrium and common atrium; chromosome complement triploid with 24 metacentric chromosomes. The other new species, Dugesiaverrucula Chen & Dong, sp. nov., is characterised by the large size of the living worm, usually exceeding 3.5 cm in length; asymmetrical openings of the oviducts; subterminal opening of ventrally displaced ejaculatory duct; vasa deferentia symmetrically opening into the postero-lateral portion of the seminal vesicle; well-developed duct between the seminal vesicle and diaphragm; single dorsal bump near the root of the penis papilla; bursal canal with pleated wall and spacious posterior section; unstalked cocoons; chromosome complement diploid with 16 metacentric chromosomes. Inter-specific molecular distances and their positions in the phylogenetic tree reveal that D.circumcisa and D.verrucula are clearly separated from their congeners.
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Autophagy-related gene 6 (Atg6) plays an essential role in autophagy, and loss of its function impairs neurogenesis. Planarian is a good model for the study of the central nervous system (CNS) regeneration. It can regenerate a new head de novo in 1 week following decapitation. Therefore, functional analysis of Atg6 in planarian CNS regeneration is very important for understanding of autophagy in the regulation of neurogenesis. In this work, we reported the molecular characteristics of Atg6 in Dugesia japonica (DjAtg6) for the first time and examined its function by RNAi. The full-length cDNA of DjAtg6 is 1366 bp encoding 423 amino acids. The deduced amino sequence of DjAtg6 contains the coil-coil domain and ß-α-repeated autophagy-specific domain shared by ATG6/Beclin 1 family. Following amputation before and after the pharynx, DjAtg6 transcripts increased and were mainly distributed in the newly regenerated brain structure. RNAi-DjAtg6 delayed planarian head regeneration with a small size of brain, and decreased the expression levels of neural-related genes. In addition, our results revealed that RNAi-DjAtg6 did not affect the stem cell proliferation, but down-regulated the cell migration-related genes mmp1 and mmp2. Furthermore, RNAi-mmp1 and RNAi-mmp2 delayed planarian head regeneration. Therefore, our results suggest that DjAtg6 is important for planarian CNS regeneration. The abnormal CNS regeneration caused by RNAi-DjAtg6 may be related to cell migration, but the detailed mechanism needs to be further investigated.
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Planárias , Animais , Autofagia , Encéfalo , Sistema Nervoso Central , Planárias/genética , Interferência de RNARESUMO
Two new species of the genus Dugesia (Platyhelminthes, Tricladida, Dugesiidae) from Hainan Island of China are described on the basis of morphological, karyological and molecular data. Dugesia semiglobosa Chen & Dong, sp. nov. is mainly characterized by a hemispherical, asymmetrical penis papilla with ventrally displaced ejaculatory duct opening terminally at tip of penis papilla; vasa deferentia separately opening into mid-dorsal portion of intrabulbar seminal vesicle; two diaphragms in the ejaculatory duct; copulatory bursa formed by expansion of bursal canal, lined with complex stratified epithelium, which projects through opening in bursa towards intestine, without having open communication with the gut; mixoploid chromosome complement diploid (2n = 16) and triploid (3n = 24), with metacentric chromosomes. Dugesia majuscula Chen & Dong, sp. nov. is mainly characterized by oviducts opening asymmetrically into female reproductive system; hyperplasic ovaries; expanded posterior section of bursal canal; vasa deferentia separately opening into mid-dorsal portion of seminal vesicle; asymmetrical penis papilla due to ventral course of ejaculatory duct, which has subterminal and dorsal opening at tip papilla; mixoploid chromosome complement diploid (2n = 16) and triploid (3n = 24); chromosomes metacentric. Apart from their anatomy, separate species status of the two new species is supported also by their genetic distances and by their positions in the phylogenetic tree. The sexualization process may have been induced by the lower temperatures, in comparison with their natural habitat, under which the worms were cultured in the laboratory.
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OBJECTIVE: To determine diagnostic performance of diffusion-weighted (DW) magnetic resonance (MR) volume and apparent diffusion coefficient values (ADCs) for assessing lymph node metastases (LNM) and good response after chemoradiation therapy (CRT) in patients with locally advanced rectal cancer (LARC). MATERIALS AND METHODS: This retrospective study consisted of 61 patients with LARC who underwent pre- and post-CRT DW images. Two radiologists independently placed free-hand regions of interest in each tumor-containing section on DW images to calculate pre- and post-CRT tumor volume and tumor volume reduction rates (Δvolume). Regions of interest were drawn to include tumor on maximum cross-sectional slice to obtain ADCs. Areas under the receiver operating characteristic curves (AUCs) were calculated to evaluate diagnostic performance in identifying LNM and good response after CRT using these parameters. RESULTS: Inter-observer agreement and intra-observer agreement were excellent for pre- and post-CRT DW MR volume (intraclass correlation coefficient [ICC], 0.889-0.948) and moderate for pre- and post-CRT ADCs (ICC, 0.535-0.811). AUCs for identifying LNM were 0.508 for pre-CRT DW MR volume versus 0.705 for pre-CRT ADC, 0.855 for post-CRT DW MR volume versus 0.679 for post-CRT ADC, and 0.887 for Δvolume versus 0.533 for ΔADC. AUCs for identifying good response were 0.518 for pre-CRT volume versus 0.506 for pre-CRT ADC, 0.975 for post-CRT volume versus 0.723 for post-CRT ADC, and 0.987 for Δvolume versus 0.655 for ΔADC. CONCLUSION: DW MR Δvolume provided high diagnostic performance in discriminating LNM after CRT. DW MR Δvolume was equally as accurate as post-CRT DW MR volume for evaluating good response. KEY POINTS: ⢠Inter-observer agreement and intra-observer agreement were excellent for pre- and post-CRT DW MR volume (intraclass correlation coefficient [ICC], 0.889-0.948) and moderate for pre- and post-CRT ADCs (ICC, 0.535-0.811). ⢠DW MR Δvolume provided high diagnostic performance in identifying LNM after CRT (AUC, 0.887) and good response (AUC, 0.987) and was significantly more accurate than pre-CRT DW MR volume (AUC, 0.508 and 0.518, respectively) and ADCs (AUC, 0.705 and 0.506, respectively). ⢠DW MR Δvolume (AUC, 0.987) was equally as accurate as post-CRT DW MR volume (AUC, 0.975) for evaluating good response, while pre-CRT DW MR volume and ADCs were not reliable for evaluating LNM and good response after CRT (AUC, 0.506-0.723).
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Neoplasias Retais , Estudos Transversais , Imagem de Difusão por Ressonância Magnética , Humanos , Metástase Linfática , Espectroscopia de Ressonância Magnética , Terapia Neoadjuvante , Neoplasias Retais/diagnóstico por imagem , Neoplasias Retais/terapia , Estudos Retrospectivos , Resultado do TratamentoRESUMO
This study is to investigate optimum apparent diffusion coefficient (ADC) parameter for predicting lymphovascular invasion (LVI), lymph node metastasis (LNM) and histology type in resectable rectal cancer. 58 consecutive patients with resectable rectal cancer were retrospectively identified. The minimum, maximum, average ADC and ADC difference value were obtained on ADC maps. Maximum ADC and ADC difference value increased with the appearance of LVI (r = 0.501 and 0.495, P < 0.001, respectively) and development of N category (r = 0.615 and 0.695, P < 0.001, respectively). ADC difference value tended to rise with lower tumor differentiation (r = - 0.269, P = 0.041). ADC difference value was an independent risk factor for predicting LVI (odds ratio = 1.323; P = 0.005) and LNM (odds ratio = 1.526; P = 0.005). Maximum ADC and ADC difference value could distinguish N0 from N1 category, N0 from N1-N2, N0-N1 from N2 (all P < 0.001). Only ADC difference value could distinguish histology type (P = 0.041). ADC difference value had higher area under the receiver operating characteristic curve than maximum ADC in identifying LVI (0.828 vs 0.797), N0 from N1 category (0.947 vs 0.847), N0 from N1-N2 (0.935 vs 0.874), and N0-N1 from N2 (0.814 vs 0.770). ADC difference value may be superior to the other ADC value parameters to predict LVI, N category and histology type of resectable rectal cancer.
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Imagem de Difusão por Ressonância Magnética , Neoplasias Retais/diagnóstico , Neoplasias Retais/cirurgia , Adulto , Idoso , Idoso de 80 Anos ou mais , Biópsia , Feminino , Neoplasias Gastrointestinais/diagnóstico , Neoplasias Gastrointestinais/patologia , Neoplasias Gastrointestinais/cirurgia , Humanos , Interpretação de Imagem Assistida por Computador , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Variações Dependentes do Observador , Razão de Chances , Prognóstico , Curva ROC , Neoplasias Retais/patologia , Estudos Retrospectivos , Fatores de Risco , Adulto JovemRESUMO
As the worldwide top-selling herbicide, glyphosate is ubiquitously distributed in the natural environment, and its influence on the ecological safety and human health has being increasingly concerned. In this study, mRNA expressions of GPX and three heat shock protein genes in freshwater planarian Dugesia japonica in response to glyphosate were determined, and two oxidative stress parameters were measured. The results suggested that GPX activity can be used as a more sensitive biomarker in contrast with GPX gene expression, and mRNA expressions of Hsp70, Hsp90 genes are more sensitive than Hsp40 for planarians in response to glyphosate stress. Besides, the deduced T-AOC as well as varied GPX activity and mRNA expression levels of Hsps also indicated that glyphosate exposure would inhibit antioxidation and induce oxidative stress in D. japonica, while specific antioxidant systems and stress proteins tried to protect cells by their own regulation. The results of this study will be helpful to elucidate the stress response mechanisms of freshwater planarians to herbicide glyphosate.
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Expressão Gênica/efeitos dos fármacos , Glicina/análogos & derivados , Herbicidas/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Planárias/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Animais , Antioxidantes/metabolismo , Biomarcadores/metabolismo , Glicina/toxicidade , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo , Estresse Oxidativo/genética , Planárias/genética , Planárias/metabolismo , GlifosatoRESUMO
BACKGROUND: Pulmonary artery aneurysm (PAA), usually associated with congenital heart disease (CHD), is a very rare clinical condition. Pulmonary artery dissection (PAD) is considered the most life-threatening complication of PAA, and patients can progress from being asymptomatic to sudden death. We report the following case of PAA associated with complicated congenital heart disease and simultaneous chronic PAD. To our knowledge, few such complicated cases have ever been reported. CASE PRESENTATION: A 36-year-old male presented to our hospital with a 10-year history of heart fatigue after activities and aggravated for 2 years. Computed tomography angiography (CTA) and echocardiogram showed a giant main pulmonary artery aneurysm (14 cm) with complicated congenital heart disease (a small patent ductus arteriosus, ventricular septal defects, aortic coarctation). Chronic PAD, which was mistaken for a pulmonary valve before operation, was detected during surgery. CONCLUSIONS: PAA associated with complicated CHD and simultaneous PAD is very rare, and its clinical symptoms are varied. Radiologists and surgeons should pay attention to determining whether this patient simultaneously had PAD when PAA was detected on preoperative CTA imaging.
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Dissecção Aórtica/complicações , Cardiopatias Congênitas/complicações , Artéria Pulmonar/cirurgia , Adulto , Dissecção Aórtica/diagnóstico por imagem , Dissecção Aórtica/cirurgia , Doença Crônica , Angiografia por Tomografia Computadorizada , Ecocardiografia , Cardiopatias Congênitas/diagnóstico por imagem , Cardiopatias Congênitas/cirurgia , Humanos , Masculino , Artéria Pulmonar/diagnóstico por imagemRESUMO
As the top-selling herbicide in the world, glyphosate distributes widely in natural environment and its influence on the ecological security and human health has attracted more and more concern. Glutathione S-transferases (GSTs) are a well-characterized superfamily of isoenzymes for cellular defense against exogenous toxic substances and therefore protect organisms from injury. In this study, the complete cDNA sequence of GST gene (named as Dja-GST) in freshwater planarian Dugesia japonica was firstly cloned by means of RACE method. The full-length Dja-GST comprises of 706 nucleotides which encodes a polypeptide of 200 amino acids. Dja-GST has two representative GST domains at the N- and C-termini. The conservative GST-N domain includes G-site Y8, F9, R14, W39, K43, P52 and S64, while the variable GST-C domain contains H-site K104, V156, D159 and L161. Sequence analysis, phylogenetic tree reconstruction and multiple alignment collectively indicate that Dja-GST belongs to the Sigma class of GST superfamily. Also, GST gene expression profile, GST enzymatic activity and MDA content in response to glyphosate exposure were systematically investigated and the correlations among them were analyzed. The results suggest that glyphosate exposure modified the mRNA transcription and enzymatic activity of GST, as well as the MDA content in planarians, indicating that Dja-GST might play an important part in organisms defending against oxidative stress induced by glyphosate. This work lays a molecular foundation for further exploring the exact functions of Dja-GST and gives an important implication for evaluating the ecological environment effects of herbicide glyphosate.
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Glutationa Transferase/genética , Glicina/análogos & derivados , Herbicidas/toxicidade , Planárias/fisiologia , Poluentes Químicos da Água/toxicidade , Animais , Clonagem Molecular , Água Doce , Glicina/toxicidade , Estresse Oxidativo , GlifosatoRESUMO
Heavy metal pollution is a global health issue affecting people worldwide, and the exploration of sensitive biomarkers to assess the toxicity of heavy metals is an important work for researchers. Cathepsin L, role as a tissue-specific biomarker to assess the biological effects of environmental pollutants, has not received much attention. In this work, the full-length cDNA of cathepsin L gene from the planarian Dugesia japonica (designated DjCatL) was cloned by rapid amplification of cDNA ends (RACE) technique. The cDNA sequence of DjCatL is 1161 bp, which encodes a protein of 346 amino acids with a molecular weight of 39.03â¯kDa. Sequence analysis revealed that DjCatL contains highly conserved ERF/WNIN, GNFD, and GCXGG motifs, which are the features of the cathepsin L protein family. Whole-mount in situ hybridization (WISH) results revealed that the transcripts of DjCatL are specifically distributed in the intestinal system, suggesting that this gene is related to food digestion in planarians. Both quantitative polymerase chain reaction (qPCR) and WISH results revealed that the transcriptional levels of DjCatL are inhibited significantly by heavy metal (Cd2+, Hg2+, and Cu2+) exposure in a dose-dependent manner. Therefore, we proposed that cathepsin L can be used as a tissue-specific biomarker to assess the heavy metal pollution in the aquatic environment.
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Catepsina L/genética , Expressão Gênica/efeitos dos fármacos , Metais Pesados/toxicidade , Planárias/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Sequência de Aminoácidos , Animais , Catepsina L/metabolismo , Clonagem Molecular , Biomarcadores Ambientais/efeitos dos fármacos , Planárias/genética , RNA Mensageiro/genéticaRESUMO
OBJECTIVE. The purpose of this study was to assess whether MR volumetric data on DW and T2-weighted MR images are correlated with lymphovascular invasion and lymph node metastases in resectable rectal cancer. MATERIALS AND METHODS. This retrospective study consisted of 50 consecutive patients with rectal cancer who underwent radical surgery within 1 week of MRI. The gross tumor volume was determined on both diffusion-weighted and T2-weighted MR images and correlated with pathologic lymphovascular invasion and lymph node metastases using univariate, multivariate, and ROC curve analyses. RESULTS. Both gross tumor volume values showed correlations with lymphovascular invasion (r = 0.750 vs r = 0.710; p < 0.0001) and lymph node metastases (r = 0.780 vs r = 0.755; p < 0.0001). Both values were associated with lymphovascular invasion and lymph node metastases in univariate analysis (all p < 0.0001), whereas only the DWI-based value was an independent risk factor for lymphovascular invasion (odds ratio = 1.207; p = 0.005) and lymph node metastases (odds ratio = 1.420; p = 0.005) in multivariate analysis. Both values could distinguish between N0 and N1, N0 and N1-N2, and N0-N1 and N2 disease (all p < 0.0001) in the Mann-Whitney U test. The area under the ROC curve was higher for the DWI-based value in lymphovascular invasion (0.899 vs 0.877), N0 vs N1 (0.865 vs 0.827), N0 vs N1-N2 (0.934 vs 0.911), and N0-N1 vs N2 (0.932 vs 0.927). CONCLUSION. Tumor volumetry data correlated with both lymphovascular invasion and lymph node metastases in resectable rectal cancer. In particular, the DWI-based gross tumor volume showed the most potential for noninvasive preoperative evaluation of lymphovascular invasion and lymph node metastases.
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As an important antioxidant enzyme, the superoxide dismutase (SOD) can protect aerobic organisms from oxidative damage through catalyzing the dismutation of superoxide into hydrogen peroxide and oxygen. The SODs have been cloned in some species and their dynamic expression or enzymatic activity in response to environmental stressors were investigated. In the current study, the full-length cDNA of two SODs from freshwater planarian Dugesia japonica were firstly cloned (named as DjCuZnSOD and DjMnSOD, respectively). The complete cDNA of DjCuZnSOD consists of 661 nucleotides encoding 186 amino acids while the 765 bp DjMnSOD encodes a polypeptide of 226 residues. Sequence analysis and multiple alignment showed that DjCuZnSOD possesses two CuZnSOD family signature motifs and an N-terminal signal peptide suggesting it is an extracellular secretory protein. DjMnSOD possesses the MnSOD family signature sequence and is predicted to be located in mitochondrion with a mitochondrial targeting sequence. Phylogenetic analysis based on CuZnSOD and MnSOD orthologs from representative species further verified that DjCuZnSOD is an extracellular CuZnSOD while DjMnSOD is a mitochondrial MnSOD. For the purpose of studying their potential role against environmental pollutants, D. japonica were exposed to glyphosate or 1-decyl-3-methylimidazolium bromide ([C10mim]Br), and the mRNA expression levels of DjCuZnSOD and DjMnSOD along with total SOD activity were measured. The results showed that DjCuZnSOD exhibited more sensitive expression profiles in response to environmental pollutants in contrast with DjMnSOD, and the total SOD activity in response to both pollutants was more related to the expression level of DjCuZnSOD than to DjMnSOD, indicating that the mRNA expression of CuZnSOD would be a more sensitive biomarker than MnSOD in monitoring the pollution of aquatic environment and CuZnSOD might play more important role than MnSOD in eliminating superoxide anions caused by pollutants in D. japonica.
Assuntos
DNA Complementar/genética , Água Doce , Regulação da Expressão Gênica/efeitos dos fármacos , Planárias/enzimologia , Planárias/genética , Superóxido Dismutase-1/metabolismo , Superóxido Dismutase/metabolismo , Poluentes Químicos da Água/toxicidade , Sequência de Aminoácidos , Animais , Clonagem Molecular , Filogenia , Planárias/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Homologia de Sequência de Aminoácidos , Superóxido Dismutase/química , Superóxido Dismutase/genética , Superóxido Dismutase-1/química , Superóxido Dismutase-1/genética , Fatores de TempoRESUMO
Catalase (CAT) is an important antioxidant enzyme that protects aerobic organisms against oxidative damage by degrading hydrogen peroxide to oxygen and water. CAT mRNAs have been cloned from many species and employed as useful biomarkers of oxidative stress. In the present study, we cloned the cDNA sequence of CAT gene from freshwater planarian Dugesia japonica (designated as DjCAT) by means of RACE method. Sequence analysis and multiple alignment jointly showed that the full-length cDNA sequence consists of 1734 nucleotides, encoding 506 amino acids. Three catalytic amino acid residues of His71, Asn144 and Tyr354, two CAT family signature sequences of a proximal active site signature (60FDRERIPERVVHAKGGGA77) and a heme-ligand signature motif (350RLFSYRDTQ358) are highly conserved, suggesting that the DjCAT belongs to the NADPH and heme-binding CAT family and has similar functions. In addition, the transcriptional level of CAT gene and activity of CAT enzyme upon acute exposure of environmental pollutants glyphosate and 1-decyl-3-methylimidazolium bromide ([C10mim]Br) were investigated systematically. The variation of CAT mRNA expression in D. japonica was quantified by real-time PCR and the results indicated that it was up-regulated after exposure to glyphosate or [C10mim]Br with a dose-dependent manner but not linearly. Even though the variation trend of CAT activity upon glyphosate stress was not monotonously increased and inconsistent with that after [C10mim]Br exposure on day 1 and 3 sampling time, with the duration prolonged to day 5 they both presented a dose-dependent increase and the differences achieved extreme significance in all treated groups compared to the control. These findings suggested that DjCAT plays an important role in antioxidant defense in D. japonica, and the mRNA expression of CAT would also be used as an effective biomarker to monitor the pollution in aquatic environment just like its corresponding enzyme.
Assuntos
Catalase/genética , Catalase/metabolismo , DNA Complementar/metabolismo , Poluentes Ambientais/farmacologia , Expressão Gênica/efeitos dos fármacos , Planárias/enzimologia , Sequência de Aminoácidos , Animais , Antioxidantes/metabolismo , Biomarcadores/metabolismo , Brometos/farmacologia , Clonagem Molecular , Relação Dose-Resposta a Droga , Glicina/análogos & derivados , Glicina/farmacologia , Herbicidas/farmacologia , Imidazóis/farmacologia , Oxirredução , Estresse Oxidativo , RNA Mensageiro/metabolismo , Análise de Sequência de DNA , Regulação para Cima/efeitos dos fármacos , GlifosatoRESUMO
OBJECTIVE: To determine whether gross tumor volume (GTV) and the maximum diameter of resectable cervical cancer at magnetic resonance imaging (MRI) could predict lymph node metastasis (LNM) and lymphovascular space invasion (LVSI). MATERIALS AND METHODS: A total of 315 consecutive patients with cervical cancer were retrospectively identified. Gross tumor volume and the maximum diameter of tumor were evaluated on MRI. Univariate and multivariate logistic regression analyses were performed to determine whether tumor size could predict LNM and LVSI. Cutoffs of GTV, maximum diameter, and the International Federation of Gynecology and Obstetrics (FIGO) classification of tumor were first investigated in 255 patients (group A) and then validated in an independent cohort of 60 patients (group B) using area under the receiver operating characteristic curve (AUC) analysis for predicting the presence of LNM and LVSI. RESULTS: Univariate analysis showed that GTV and the maximum diameter of tumor could predict LNM and LVSI (all P < 0.0001). Multivariate analyses indicated GTV as an independent risk factor of LNM and LVSI (all P < 0.0001). In group A, GTV, the maximum diameter, and the FIGO stage could identify LNM (AUC, 0.813, 0.741, and 0.69, respectively) and LVSI (AUC, 0.806, 0.751, and 0.684, respectively). In group B, GTV, the maximum diameter, and the FIGO stage could help identify LNM (AUC, 0.902, 0.825, and 0.759, respectively) and LVSI (AUC, 0.771, 0.748, and 0.700, respectively). CONCLUSIONS: Gross tumor volume and the maximum diameter of resectable cervical cancer at MRI demonstrated capability in predicting LNM and LVSI, which were more accurate than FIGO stage.