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Objective: To explore the clinical value of ultrasound-guided radiofrequency ablation in the treatment of retroperitoneal tumors. Methods: The clinical data of 13 patients with retroperitoneal tumors treated with ultrasound-guided radiofrequency ablation in the First Affiliated Hospital of Zhengzhou University from January 2018 to January 2020 were analyzed retrospectively. The ablation effect was evaluated and the postoperative complications were observed. The changes of tumor volume before and after radiofrequency ablation were compared. Results: The symptoms of pain and dyspepsia were significantly improved after radiofrequency ablation, and the hospital stay was (9.2±2.9) days. The tumor was ablated completely in 10 cases, tumor residual in 1 case and tumor metastasis in 2 cases. One patient had postoperative duodenal perforation complicated with intra-abdominal infection, and no serious complications occurred in other patients. There were 20 lesions in 13 patients. The maximum diameter of 20 lesions before operation and 1, 3, 6 months after operation were (39.5±15.9) mm, (30.6±4.9)mm, (15.6±7.7) mm and (9.9±3.1) mm, respectively, the maximum diameters of 1, 3 and 6 months after operation were smaller than that before operation (P<0.05). Conclusion: Ultrasound-guided radiofrequency ablation is a real-time, accurate, safe and effective minimally invasive treatment with few complications, and has a high clinical value for retroperitoneal tumors.
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Ablación por Catéter , Ablación por Radiofrecuencia , Neoplasias Retroperitoneales , Humanos , Neoplasias Retroperitoneales/diagnóstico por imagen , Neoplasias Retroperitoneales/cirugía , Estudios Retrospectivos , Resultado del Tratamiento , Ultrasonografía IntervencionalRESUMEN
The article "Circular RNA circCRIM1 suppresses lung adenocarcinoma cell migration, invasion, EMT, and glycolysis through regulating miR-125b-5p/BTG2 axis, by S.-J. Zhang, J. Ma, J.-C. Wu, Z.-Z. Hao, Y.-A. Zhang, Y.-J. Zhang, published in Eur Rev Med Pharmacol Sci 2020; 24 (7): 3761-3774-DOI: 10.26355/eurrev_202004_20841-PMID: 32329853" has been withdrawn from the authors due to some technical reasons. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/20841.
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OBJECTIVE: The present studies indicate that circRNAs play pivotal roles in human cancers. Lung adenocarcinoma (LUAC), one of lung cancer types, has high metastasis rate. Herein, we focused our study on the function and mechanism of circular RNA circCRIM1 in LUAC development. PATIENTS AND METHODS: Quantitative Real-time polymerase chain reaction (qRT-PCR) was performed to detect the levels of circCRIM1, miR-125b-5p, and BTG anti-proliferation factor 2 (BTG2). Transwell assay was carried out to assess cell migration and invasion. The protein levels of BTG2, EMT markers, and HK2 were measured by Western blot. Glycolysis was analyzed through determining glucose consumption and lactate production. Furthermore, the targets of circCRIM1 and miR-125b-5p were predicted and verified by starBase and the dual-luciferase reporter assay, respectively. Also, whether circCRIM1 affecting tumor growth in vivo was explored using mouse xenograft assay. RESULTS: CircCRIM1 and BTG2 were downregulated, and miR-125b-5p was upregulated in LUAC tissues/cells. CircCRIM1 upregulation inhibited LUAC cell migration, invasion, epithelial-mesenchymal transition (EMT), glycolysis, and tumor growth. Moreover, circCRIM1 regulated LUAC cell development through targeting miR-125b-5p. MiR-125b-5p affected LUAC cell growth via binding to BTG2. Also, circCRIM1 promoted BTG2 expression by inhibiting miR-125b-5p expression in LUAC cells. CONCLUSIONS: CircCRIM1 was lowly expressed in LUAC. Moreover, circCRIM1 functioned as a sponge of miR-125b-5p to improve BTG2 expression, thereby suppressing LUAC development. Our finding indicated that circCRIM1 could be considered as a biomarker and target for the diagnosis and therapy of LUAC patients.
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Adenocarcinoma del Pulmón/metabolismo , Movimiento Celular , Transición Epitelial-Mesenquimal , Glucólisis , Proteínas Inmediatas-Precoces/metabolismo , Neoplasias Pulmonares/metabolismo , MicroARNs/metabolismo , ARN Circular/metabolismo , Proteínas Supresoras de Tumor/metabolismo , Adenocarcinoma del Pulmón/patología , Proliferación Celular , Células Cultivadas , Humanos , Proteínas Inmediatas-Precoces/genética , Neoplasias Pulmonares/patología , MicroARNs/genética , Invasividad Neoplásica , ARN Circular/genética , Proteínas Supresoras de Tumor/genéticaRESUMEN
AIM: This study study aimed to investigate the effects of explicit and tacit knowledge sharing on clinical decision-making abilities and the mediating role of trust among registered nurses at Korean hospitals. BACKGROUND: Decision-making abilities comprise a key area of nursing practice and link nurses' perceptions with behaviours. INTRODUCTION: Tacit knowledge is embedded within an individual and cannot be expressed or transmitted to other people in a specific form. Over time, new nurses gradually gain experience and tacit knowledge and become experts. Trust, an organizational characteristic, may serve as a potential mediator in the association between knowledge sharing and decision-making abilities among nurses. However, few studies have investigated the mediatory role of trust in this association. METHOD: The data were collected from 210 nurses selected via random sampling. The research instrument in the model included Knowledge-Sharing Behavior, Trust, and Clinical Decision-Making in Nursing Scale. Structural equation modelling was used to analyse the collected data. FINDINGS: The study findings showed that explicit knowledge sharing directly affects decision-making abilities, whereas tacit knowledge sharing is only associated with decision-making abilities when trust plays a mediating role. DISCUSSION: A higher level of organizational trust can improve clinical decision-making abilities via tacit knowledge sharing. CONCLUSION: This study demonstrated that unlike explicit knowledge, which is shared more easily, tacit knowledge sharing does not directly lead to clinical decision-making abilities. A higher level of organizational trust leads to a stronger beneficial effect of tacit knowledge sharing on clinical decision-making abilities. IMPLICATIONS FOR NURSING AND HEALTH POLICY: These findings concerning the mediatory role of trust on the association between knowledge sharing and clinical decision-making abilities provide new knowledge that will allow nurses, managers, and researchers to support the clinical decision-making abilities of nurses.
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Conducta Cooperativa , Relaciones Interprofesionales , Personal de Enfermería en Hospital/psicología , Confianza/psicología , Compromiso Laboral , Actitud del Personal de Salud , Humanos , Gestión del Conocimiento , Personal de Enfermería en Hospital/organización & administración , Cultura OrganizacionalRESUMEN
Radiation-induced sterile insect technique is a biologically based, environment-friendly method for the suppression or eradication of a number of insect pests. Although the basic mechanisms underlying the technology have been well studied, little is known about the cell responses in organisms. Characterization of the metabolic shift associated with radiation exposure in sterile insects would be helpful for understanding the detailed mechanism underlying this technique and promote its practical application. In this article, a metabolomic study was performed to characterize the global metabolic changes induced by radiation using untreated and 40 Gy (60)Coγ-irradiated testes of Japanese pine sawyer, Monochamus alternatus Hope. Differential metabolites were detected and tentatively identified. Many key metabolites in glycolysis and the tricarboxylic acid cycle, as well as most fatty and amino acids, were elevated in irradiated male M. alternatus, presumably resulting from depression of glycolysis and the tricarboxylic acid cycle, each of which are important pathways for energy generation Adenosine Triphosphate (ATP) in insect spermatozoa. The findings in this article will contribute to our knowledge of the characteristic metabolic changes associated with irradiation sterility and understand the molecular mechanisms underlying radiation-induced sterile insect technique.
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Metabolismo de los Hidratos de Carbono/efectos de la radiación , Escarabajos/efectos de la radiación , Metabolismo de los Lípidos/efectos de la radiación , Metaboloma/efectos de la radiación , Control Biológico de Vectores , Animales , Escarabajos/metabolismo , Cromatografía de Gases y Espectrometría de Masas , Infertilidad Masculina , MasculinoRESUMEN
In an effort to identify novel biallelically inactivated tumor suppressor genes (TSGs) in sporadic invasive and preinvasive non-small-cell lung cancer (NSCLC) genomes, we applied a comprehensive integrated multiple 'omics' approach to investigate patient-matched, paired NSCLC tumor and non-malignant parenchymal tissues. By surveying lung tumor genomes for genes concomitantly inactivated within individual tumors by multiple mechanisms, and by the frequency of disruption in tumors across multiple cohorts, we have identified a putative lung cancer TSG, Eyes Absent 4 (EYA4). EYA4 is frequently and concomitantly deleted, hypermethylated and underexpressed in multiple independent lung tumor data sets, in both major NSCLC subtypes and in the earliest stages of lung cancer. We found that decreased EYA4 expression is not only associated with poor survival in sporadic lung cancers but also that EYA4 single-nucleotide polymorphisms are associated with increased familial cancer risk, consistent with EYA4s proximity to the previously reported lung cancer susceptibility locus on 6q. Functionally, we found that EYA4 displays TSG-like properties with a role in modulating apoptosis and DNA repair. Cross-examination of EYA4 expression across multiple tumor types suggests a cell-type-specific tumorigenic role for EYA4, consistent with a tumor suppressor function in cancers of epithelial origin. This work shows a clear role for EYA4 as a putative TSG in NSCLC.
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Carcinoma de Pulmón de Células no Pequeñas/epidemiología , Silenciador del Gen , Neoplasias Pulmonares/patología , Transactivadores/genética , Carcinoma de Pulmón de Células no Pequeñas/genética , Cromosomas Humanos Par 6 , Metilación de ADN , Epigénesis Genética , Eliminación de Gen , Regulación Neoplásica de la Expresión Génica , Frecuencia de los Genes , Genes Supresores de Tumor , Estudios de Asociación Genética , Variación Genética , Genoma Humano , Humanos , Neoplasias Pulmonares/genética , Polimorfismo de Nucleótido Simple , Transactivadores/metabolismo , Células Tumorales CultivadasRESUMEN
BACKGROUND: Limited physiological data for Tibetan macaques are available at present. This study will provide more rationale for evaluating this species. METHODS: Thirty-seven Tibetan macaques (15 males and 22 females) were used in this study. Somatometric measurements, clinical chemistry and hematology parameters, insulin, and C-peptide were analyzed. RESULTS: Females had higher values of waist and waist hip ratio (WHR) than males in somatometric measurements. There were no significant differences between the two genders in hematology. Significant differences between males and females were only found for aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in biochemistry testing. In addition, females had higher fasting insulin and C-peptide than males. There was a strongly positive correlation between age and some somatometric parameters. CONCLUSIONS: These physiological data will provide veterinarians and researchers with baseline values to evaluate experimental results using Tibetan macaques.
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Macaca/anatomía & histología , Macaca/sangre , Factores de Edad , Alanina Transaminasa/sangre , Animales , Aspartato Aminotransferasas/sangre , Biometría , Recuento de Células Sanguíneas , Pruebas de Coagulación Sanguínea , Péptido C/sangre , Electrólitos/sangre , Femenino , Hemoglobina Glucada/análisis , Hidrocortisona/sangre , Insulina/sangre , Masculino , Valores de ReferenciaRESUMEN
The ontogeny of IgM-producing cells was studied in juvenile mandarin fish Siniperca chuatsi, an important fish in China's aquaculture sector. The IgM-producing cells were localised through in situ hybridisation with a probe complementary to the Ig mu-chain in lymphoid-related tissues, including head kidney, spleen, thymus, intestine and gills. In head kidney, transcripts of Ig mu were first detected at 20days post-hatching (dph) with a few positive signals, and the number of IgM-producing cells increased obviously from 39dph onwards. At 136dph, a large amount of positive cells were observed in the entire organ with clusters of these cells located around the blood vessels. In spleen, IgM-producing cells were found from 26dph onwards, followed by an increase until 67dph; clusters of positive cells were also detected around blood vessels at 102dph. In thymus, IgM-producing cells were first observed at 39dph; thereafter, no obvious increase was detected until 78dph. The positive cells in thymus were distributed mainly in the outer zone of thymus. A few IgM-producing cells were still observed in thymus of 1-year-old mandarin fish. IgM-producing cells were not detected in the intestine until 87dph, with several discrete positively stained cells distributed in the lamina propria. IgM-producing cells, scattered mainly in primary gill filaments around blood vessels, were detected in gills from 90dph. As in other teleosts, these results indicated that the head kidney appears to be the primary organ for IgM production in mandarin fish, and IgM-producing cells exist in all organs examined in the present study, implying their lymphoid role in fish. In addition, it is suggested that vaccination after 20dph may be much more effective in mandarin fish.
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Inmunoglobulina M/biosíntesis , Inmunoglobulina M/genética , Tejido Linfoide/inmunología , Perciformes/genética , Perciformes/inmunología , Animales , Células Productoras de Anticuerpos/inmunología , Secuencia de Bases , ADN Complementario/genética , Branquias/citología , Branquias/inmunología , Cadenas mu de Inmunoglobulina/biosíntesis , Cadenas mu de Inmunoglobulina/genética , Hibridación in Situ , Intestinos/citología , Intestinos/inmunología , Riñón/citología , Riñón/inmunología , Tejido Linfoide/citología , Tejido Linfoide/crecimiento & desarrollo , Perciformes/crecimiento & desarrollo , Bazo/citología , Bazo/inmunología , Timo/citología , Timo/inmunologíaRESUMEN
BACKGROUND: This study aimed to retrospectively analyze the effect of long-term streptozotocin-induced diabetes mellitus (STZ-DM) on adolescent cynomolgus monkeys. METHODS: A total of 12 monkeys (six STZ-DM and six controls) were monitored for fasting glucose levels and locomotor activities, tested for hematological and serum parameters, measured for body weight and somatometric values. RESULTS: Fasting glucose was maintained at high levels in STZ-DM monkeys. At the age when normal adolescent monkeys dramatically increased their weight, STZ-DM led to the retardation of weight increase in diabetic monkeys. Moreover, STZ-DM monkeys showed abnormal lipid levels and somatometric measurements. In locomotor activity test, STZ-DM monkeys were more active than control ones. CONCLUSIONS: Long-term STZ-DM disrupts the normal growth of young monkeys and interferes with some aspects of hormone, lipid metabolism and physical activities. Mean plasma glucose (MPG) appeared to be an important factor in physical activity abnormalities of STZ-DM monkeys.
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Glucemia/metabolismo , Diabetes Mellitus Experimental/sangre , Macaca fascicularis/sangre , Enfermedades de los Monos/sangre , Actividad Motora , Animales , Peso Corporal , Diabetes Mellitus Experimental/fisiopatología , Macaca fascicularis/fisiología , Masculino , Enfermedades de los Monos/fisiopatología , Estudios Retrospectivos , Factores de TiempoRESUMEN
The Japanese pine sawyer Monochamus alternatus is one of the major forest pests. It damages pine directly and transfers the nematode Bursaphelenchus xylophilus to pine wood; resulting in serious economic losses around the world every year. Alpha-tubulin is one of most important proteins in most species. We cloned a ubiquitously expressed M. alternatus alpha-tubulin gene and analysed its nucleotides and protein structure; its sequence characters are consistent with what have been reported in other insects. The alignment of proteins showed that there is high homology of alpha-tubulin between M. alternatus and other species. Western blot and immunocytochemistry analyses suggested a common epitope of alpha-tubulin between M. alternatus and Strongylcentrotus purpuratus. We also expressed the protein in Escherichia coli for further functional studies.
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Escarabajos/genética , Regulación de la Expresión Génica , Tubulina (Proteína)/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Western Blotting , Clonación Molecular , Electroforesis en Gel de Poliacrilamida , Tracto Gastrointestinal/citología , Tracto Gastrointestinal/ultraestructura , Inmunohistoquímica , Datos de Secuencia Molecular , Filogenia , Estructura Secundaria de Proteína , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Alineación de Secuencia , Análisis de Secuencia de ADN , Tubulina (Proteína)/química , Tubulina (Proteína)/metabolismoRESUMEN
Early-trimester human fetal pancreas is a promising potential source of pancreatic progenitor cells. However, the ethical controversy associated with the source of these cells, and technical difficulties associated with their differentiation into insulin-producing cells have limited both their availability and utility. This study aimed to characterize a population of pancreatic progenitor cells (PPCs) isolated from human fetus and describe the effects of a novel factor, PDZ-domain containing-2 (PDZD2), and its secreted form (sPDZD2), on PPC proliferation and differentiation. In particular, we examined and characterized the expression of several stem cell (nestin, ABCG2, c-kit), growth and differentiation markers (GLP-1R, c-met, erbB1), and PDZD2 in PPCs by RT-PCR, Western blot, and immunocytochemistry. We also examined the effects of sPDZD2 on PPC proliferation and differentiation by examining BrdU incorporation, MTT, cell number, and real-time PCR as well as ELISA. PPCs were isolated, cultured and characterized from human fetal pancreas. PDZD2 and sPDZD2 were detected at high levels in both human fetal pancreas and in PPCs. sPDZD2 acted as a potent mitogen on PPCs, and inhibited the differentiation of PPC-derived islet-like cell-clusters (ICCs), evidenced by the downregulation of Isl-1, Pdx-1, and insulin mRNA levels. sPDZD2 treatment also reduced levels of C-peptide in ICCs. These results show that a novel pancreatic developmental factor, PDZD2, is sufficient to promote the proliferation of human fetal PPCs while limiting differentiation of ICCs into islet/endocrine cells. Findings from this study will contribute to the development of improved methods for islet transplantation therapy in the treatment of diabetes.
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Proteínas Adaptadoras Transductoras de Señales/fisiología , Diferenciación Celular/fisiología , Proliferación Celular , Proteínas de Neoplasias/fisiología , Páncreas/metabolismo , Proteínas Adaptadoras Transductoras de Señales/genética , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Western Blotting , Péptido C/metabolismo , Moléculas de Adhesión Celular , Diferenciación Celular/genética , Línea Celular , Humanos , Inmunohistoquímica , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Páncreas/citología , Páncreas/embriología , Proteínas Recombinantes/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
ONYX-015 is an attenuated chimeric human group C adenovirus, which preferentially replicates in and lyses tumor cells that are p53 negative. The purpose of this phase I, dose-escalation study was to determine the safety and feasibility of intravenous infusion with ONYX-015 in combination with enbrel in patients with advanced carcinoma. Enbrel is a recombinant dimer of human tumor-necrosis factor (TNF)-alpha receptor, previously shown to reduce the level of functional TNF. Nine patients, three in each cohort received multiple cycles of ONYX-015 infusion (1 x 10(10), 1 x 10(11) and 1 x 10(12) vp weekly for 4 weeks/cycle) in addition to subcutaneous enbrel (only during cycle 1) injections per FDA-indicated dosing. Of the nine patients, four had stable disease. No significant adverse events were attributed to the experimental regimen, confirming that enbrel can be safely administered along with oncolytic virotherapy. Two of the three patients in cohort 3 had detectable viral DNA at days 3 and 8 post-ONYX-015 infusion. Their detectable circulating viral DNA was markedly higher during cycle 1 (with enbrel coadministration) as compared with cycle 2 (without enbrel) at the same time points. Area under the curve determinations indicate a marked higher level of TNF-alpha induction and accelerated clearance at cycle 2 in the absence of enbrel. Further assessment is recommended.
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Adenoviridae , Antineoplásicos/efectos adversos , Carcinoma/tratamiento farmacológico , Inmunoglobulina G/efectos adversos , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Anciano , Antineoplásicos/administración & dosificación , Antineoplásicos/uso terapéutico , Etanercept , Femenino , Humanos , Inmunoglobulina G/administración & dosificación , Inmunoglobulina G/uso terapéutico , Infusiones Intravenosas , Masculino , Persona de Mediana Edad , Receptores del Factor de Necrosis Tumoral/administración & dosificación , Receptores del Factor de Necrosis Tumoral/uso terapéutico , Factor de Necrosis Tumoral alfa/sangre , Vacunas ViralesRESUMEN
To identify signature targets associated with patient-specific cancer lesions based on tumor versus normal tissue differential protein and mRNA coexpression patterns for the purpose of synthesizing cancer-specific customized RNA interference knockdown therapeutics. Analysis of biopsied tissue involved two-dimensional difference in-gel electrophoresis (2D-DIGE) analysis coupled with MALDI-TOF/TOF mass spectrometry for proteomic assessment. Standard microarray techniques were utilized for mRNA analysis. Priority was assigned to overexpressed protein targets with co-overexpressed genes with a high likelihood of functional nodal centrality in the cancer network as defined by the interactive databases BIND, HPRD and ResNet. HPLC-grade small interfering RNA (siRNA) duplexes were utilized to assess knockdown of target proteins in expressive cell lines as measured by western blot. Seven patients with metastatic cancer underwent biopsy. One patient (RW001) had biopsies from two disease sites 10 months apart. Seven priority proteins were identified, one for each patient (RACK 1, Ras related nuclear protein, heat-shock 27 kDa protein 1, superoxide dismutase, enolase1, stathmin1 and cofilin1). Prioritized proteins in RW001 from the two disease sites over time were the same. We demonstrated >80% siRNA inhibition of RACK 1 and stathmin1 of inexpressive malignant cell lines with correlated cell kill. Identification of functionally relevant target gene fingerprints, unique to an individual's cancer, is feasible 'at the bedside' and can be utilized to synthesize siRNA knockdown therapeutics. Further animal safety testing followed by clinical study is recommended.
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Genómica , Neoplasias/tratamiento farmacológico , Proteómica , Interferencia de ARN/fisiología , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Neoplasias/genética , Neoplasias/metabolismoRESUMEN
Immunoglobulin light chain cDNA sequences of a perciform fish, the mandarin fish Siniperca chuatsi were amplified from head kidney mRNA by reverse transcription (RT)-PCR and RACE methods using degenerated primer and gene specific ones. In cDNA sequences of the VL region, nucleotide exchanges were present mainly within CDRs, although a lesser degree of variability was also found in FRs. Moreover, the length of CDR1 and CDR3 in the mandarin fish is shorter than in most other fish species. In the middle of S. chuatsi CL region, a microsatellite sequence (AGC)(6-8) was found, which is also present in another perciform species, the spotted wolffish (Anarhichas minor). The comparison of amino acid sequence of the mandarin fish CL domain with those of other vertebrates showed the highest degree of similarity of 94.5% to the spotted wolffish, while the similarity with rainbow trout (Oncorhynchus mykiss) Ig L1 (62.7%) and channel catfish (Ictalurus punctatus) Ig LG (55.9%) isotypes is also higher. However, there is only 50% identity in the VL regions between the mandarin fish and the wolffish. The sequence similarity of the mandarin fish CL domain with those of higher vertebrate did not readily allow it to be classified as kappa or lambda isotype. The phylogenetic analyses also demonstrated that the CL genes of the mandarin fish and most other teleost fish cluster as a separate branch out of the mammal kappa and lambda branches.
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ADN Complementario/genética , Cadenas Ligeras de Inmunoglobulina/genética , Región Variable de Inmunoglobulina/genética , Perciformes/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Cadenas Ligeras de Inmunoglobulina/inmunología , Región Variable de Inmunoglobulina/inmunología , Datos de Secuencia Molecular , Perciformes/inmunología , Filogenia , ARN Mensajero/química , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Alineación de SecuenciaRESUMEN
The genetic structure of populations of the fish cestode, Bothriocephalus acheilognathi collected from Bailianhe Reservoir (BLH), Changshou (CSH) and Liangzi (LZH) Lakes was investigated by using 8 microsatellite loci. A total of 108 adult worms were genotyped at each of the 8 loci. For the 3 populations, the mean number of alleles per locus ranged from 2.38 to 5.5, and the mean expected heterozygosity ranged from 0.432 to 0.559. The average polymorphic information content (PIC) was from 0.384 to 0.492. The significant Fis values indicated non-random mating within LZH and BLH populations. On the other hand, when samples were further classified into subpopulations at the level of host fish species, no or little heterozygote deficiency was detected at most loci, showing that cross-fertilization, predominantly, but not exclusively, must have occurred within the subpopulations. Microsatellite markers also revealed an unexpected high level of genetic differentiation, as measured by R(st) and N(m) values or by delta(u)2 genetic distance among subpopulations from different hosts. Factors influencing the population genetic structure and the parasite host specificity are discussed.
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Cestodos/genética , Peces/parasitología , Variación Genética/genética , Repeticiones de Microsatélite/genética , Alelos , Animales , China , Polimorfismo GenéticoAsunto(s)
Cadenas Pesadas de Inmunoglobulina/genética , Inmunoglobulina M/genética , Perciformes/inmunología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , ADN Complementario/genética , Biblioteca Genómica , Datos de Secuencia Molecular , Perciformes/genética , ARN/química , ARN/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinariaRESUMEN
The molecular variation in Bothriocephalus acheilognathi Yamaguti, 1934 from 11 species of freshwater fish collected in Australia, China, the Czech Republic, England and Hawaii was investigated by determining the nucleotide sequences of the internal transcribed spacer region. The length of the first and second internal transcribed spacer sequences of multiple individuals ranged from 553 to 571 bp and 553 to 615 bp, and the G + C content from 53.1 to 53.5%. The percentage sequence divergence varied between 0 and 0.9% in the ITS1 and 0 and 6.6% in the ITS2, respectively, indicating the occurrence of intraspecific variation. It is demonstrated that the fragment length variation resulted primarily from microsatellite polymorphisms present in the ITS region, especially in the ITS2 region. Phylogenetic analyses revealed that B. acheilognathi examined in this study consisted of three closely related genotypes with certain degrees of host-specificity, and the genotype representing isolates from Cyprinus carpio L. was the most common and diverse form within the species B. acheilognathi.
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Cestodos/genética , Cyprinidae/parasitología , ADN de Helmintos/genética , ADN Espaciador Ribosómico/genética , Animales , Secuencia de Bases , Cestodos/química , Cestodos/clasificación , ADN de Helmintos/química , ADN Espaciador Ribosómico/química , Variación Genética , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa , Alineación de Secuencia , Análisis de Secuencia de ADNRESUMEN
Non-small-cell lung cancer frequently contains oncogenetic defects (mutations in ras, retinoblastoma, and p53 genes) that contribute to disease pathophysiology. Recent studies and clinical trials have focused on gene therapy approaches that either replace the function of defective tumor-suppressor genes such as p53 or inactivate mutant oncogenes such as ras. Ribozymes are RNA molecules with highly specific intrinsic enzymatic activity against target RNA sequences, which can discriminate mutant sequences that differ by a single base from their wild-type counterparts. Following binding to the RNA substrate by base-pair complementation, the ribozyme cleaves the target RNA irreversibly, then releases itself for new rounds of subsequent cleavage, resulting in significantly improved target:effector stoichiometry as compared with antisense oligonucleotides of the same specificity. Transcript-specific ribozymes have been used extensively for experimental oncogene inactivation. Ribozymes are effective for targeting mutant ras, p53, or the multidrug-resistant gene product for lung cancer cells in vitro. However, their in vivo effect is not well defined against this malignancy. We recently characterized the antitumor properties of an anti-K-ras ribozyme specific for the K-ras codon 12 mutation (GGT-->GTT). When delivered as a transgene by an adenoviral vector (ADV), the K-ras ribozyme (KRbz) suppressed growth of lung tumor xenografts expressing the relevant mutation, whereas the corresponding antisense sequence lacking catalytic activity did not. Multiple intratumoral (3-5) injections of KRbz-ADV were effective in producing complete tumor regressions of preexisting tumor xenografts. Clinical trials are under consideration to examine the applicability of this anti-K-ras ribozyme for treatment of non-small-cell lung cancers expressing the relevant mutation.
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ras mutations represent one of the most common oncogenetic lesions in human non-small cell lung cancer (NSCLC) and adversely affect the survival of patients afflicted with this disease. ras-directed gene therapy in the past employed primarily antisense oligonucleotides (AS-ODN) or expression vectors (such as a viral vector construct) that deliver the antisense sequence to inactivate the mutant oncogene message. These approaches produced minimal toxicity, and yet were limited in efficacy. Ribozymes present a viable alternative in antisense therapy by virtue of their renewable catalytic capability for site-specific RNA cleavage. We recently produced an adenoviral vector with a hammerhead ribozyme transgene (KRbz) that is specific for the K-ras codon 12 mutant sequence GUU, given the considerations that (a) in the United States, approx 30% of human NSCLCs express K-ras oncogene mutations, nearly all of which reside in codon 12; (b) anti-K-ras, anti-H, as well as anti-N-ras hammerhead ribozymes are potent growth inhibitors in various human cancers tested; and (c) in vitro and animal model studies suggest that ribozymes directed at oncogene (K- and H-ras C-fos, BCR-ABL) or human immunodeficiency viral gene messages are more effective than their antisense counterpart. This article describes the techniques involved in the production of the KRbz-adenoviral vector that is specific for the K-ras mutation GTT, and summarizes its in vivo antitumor effect against NSCLC xenografts expressing the relevant K-ras mutation in athymic mice.
Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas/genética , Genes ras , Técnicas Genéticas , Vectores Genéticos/genética , Neoplasias Pulmonares/genética , ARN Catalítico/genética , Línea Celular , Análisis Mutacional de ADN/métodos , Ácidos Grasos Monoinsaturados/química , Humanos , Liposomas , Mutación , Plásmidos , Compuestos de Amonio Cuaternario/química , Recombinación Genética , Análisis de Secuencia de ADN , Transfección/métodosRESUMEN
Cyprinid fish, Hemiculter leucisculus, Cultrichthys erythropterus and Culter dabryi, were sampled from Liangzi, Honghu and Tangxun lakes in the flood plain of the Yangtze River. The cestode Bothriocephalus acheilognathi Yamaguti, 1934 was found in the 3 lakes, but C. erythropterus sampled from Liangzi lake was found uninfected due probably to the small sample size. Findings of the cestode in the 3 lakes represent the first record of the parasite in the flood plain of the Yangtze River, indicating that B. acheilognathi may be distributed much wider in China than previously recognized.