RESUMO
The envelope protein (Env) of the Jaagsiekte sheep retrovirus (JSRV) is known to be a unique oncoprotein responsible for inducing ovine pulmonary adenocarcinoma (OPA). The objective of this study was to prepare a specific monoclonal antibody (mAb) against the JSRV Env protein using bioinformatic analysis. According to the structure and epitope prediction results of JSRV Env, the JSRV-Env572-615 antigen was prepared via peptide synthesis (amino acid sequence 572-615, denoted as JSRV-Env572-615). BALB/c mice were immunized to prepare the anti-JSRV-Env572-615 mAb. Spleen cells were fused with SP2/0 myeloma cells after being screened by indirect ELISA and cloned by limiting dilution. The specificity of mAb was evaluated by western blot analysis and immunohistochemistry assays. Western blot results showed that the JSRV Env protein was able to bind to mAb with high specificity. Immunohistochemistry assays demonstrated that the mAb was able to recognize JSRV Env in adenomatous hyperplasia of the lung. Furthermore, JSRV was detected in peripheral blood leukocytes during the pre-clinical period of OPA in 2 of the 25 sheep using this newly synthesized mAb. Therefore, this mAb may be a useful tool for the detection of JSRV in sheep.
Assuntos
Adenocarcinoma/diagnóstico , Adenocarcinoma/veterinária , Anticorpos Monoclonais/biossíntese , Anticorpos Antivirais/biossíntese , Retrovirus Jaagsiekte de Ovinos/imunologia , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/veterinária , Adenomatose Pulmonar Ovina/diagnóstico , Adenocarcinoma/imunologia , Adenocarcinoma/virologia , Adenocarcinoma de Pulmão , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/química , Anticorpos Monoclonais/isolamento & purificação , Anticorpos Antivirais/química , Anticorpos Antivirais/isolamento & purificação , Especificidade de Anticorpos , Biologia Computacional , Diagnóstico Precoce , Epitopos/química , Epitopos/imunologia , Produtos do Gene env/química , Produtos do Gene env/imunologia , Retrovirus Jaagsiekte de Ovinos/isolamento & purificação , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/virologia , Pulmão/imunologia , Pulmão/virologia , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/virologia , Camundongos , Camundongos Endogâmicos BALB C , Peptídeos/administração & dosagem , Peptídeos/síntese química , Peptídeos/imunologia , Adenomatose Pulmonar Ovina/imunologia , Adenomatose Pulmonar Ovina/virologia , Ovinos , Carneiro Doméstico , Baço/citologia , Baço/imunologiaRESUMO
The protective effect of procyanidine and its oligomers against high glucose-mediated oxidative stress injury in endothelial progenitor cells (EPCs), and effect of procyanidin on vascular endothelial growth factor receptor-2 (VEGFR-2) expression and downstream signal pathway were analyzed in vitro. Rat bone marrow mononuclear cells were isolated, cultured under normal and high glucose (HG) conditions, and the changes in cell morphology observed. The EPCs were identified, and the oxidative stress products produced by EPCs (under normal and HG conditions) were quantified. Subsequently, an appropriate number of EPCs were cultured with and without procyanidin (OPC), and the MDA concentration and relative expression of VEGFR-2, AKT, IκB-α, and nuclear factor (NF)-κB were detected 1, 3, 5, and 7 days post-culture. We observed minor (round, translucent, gradually adhering) and significant (fusiform morphology/pebble distribution) cell morphological changes 3 and 7 days post-culture, respectively. Apoptosis and oxidative stress product release in EPCs cultured with HG increased significantly compared to the control group (P < 0.05). The oxidative stress product generation and relative expression of VEGFR-2, AKT, IkB-α, and NF-κB were not significantly affected by OPC addition in normal glucose conditions (P > 0.05); alternately, products generated as a result of oxidative stress were significantly reduced, the relative expression of VEGFR-2, AKT, and NF-κB protein was upregulated, and that of IκB-α was downregulated (P < 0.05) in HG + OPC EPCs. Therefore, procyanidin may promote cell proliferation by alleviating oxidative damage to EPCs under HG conditions, and upregulating VEGFR-2 expression and its downstream signal pathway.
Assuntos
Biflavonoides/farmacologia , Catequina/farmacologia , Células Progenitoras Endoteliais/efeitos dos fármacos , Células Progenitoras Endoteliais/metabolismo , Glucose/farmacologia , Proantocianidinas/farmacologia , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Animais , Células Cultivadas , Estresse Oxidativo/efeitos dos fármacos , Ratos , Transdução de Sinais/efeitos dos fármacosRESUMO
We examined the expression of angiopoietin-2 in serum samples from patients diagnosed with colorectal cancer and healthy volunteers and investigated the feasibility of using angiopoietin-2 as a potential diagnostic colorectal cancer biomarker. Enzyme-linked immunosorbent assays were used to measure the levels of angiopoietin-2 in patients with colorectal cancer and healthy control subjects. Correlations between serum angiopoietin-2 levels and clinicopathological factors were investigated. A receiver operating characteristic curve was used to predict cut-off values of the markers. Serum concentrations of angiopoietin-2 were significantly higher in patients with colorectal cancer than in controls (2896 ± 1273 vs 1554 ± 991 pg/mL, P = 0.004). Serum angiopoietin-2 expression levels were significantly positively correlated with TNM stage (P = 0.003), lymph node involvement (P = 0.04), and distant metastases (P = 0.005). Receiver operating characteristic curve analysis revealed that serum level of angiopoietin-2 was a potential biomarker for differentiating colorectal cancer patients from controls and had a receiver operating characteristic area under the curve of 0.859 (95% confidence interval = 0.740-0.978). At a cut-off value of 2710 pg/mL, the sensitivity was 79.3% and the specificity was 82.4%. Our results suggest that angiopoietin-2 can be used as a diagnostic biomarker for colorectal cancer in clinical practice. Additional studies are needed to clarify the detailed mechanism of angiopoietin-2 in the carcinogenesis and metastasis of colorectal cancer.
Assuntos
Angiopoietina-2/sangue , Neoplasias Colorretais/sangue , Neoplasias Colorretais/diagnóstico , Adulto , Idoso , Biomarcadores , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica , Estadiamento de Neoplasias , Prognóstico , Curva ROC , Carga TumoralRESUMO
The association between the CCDC26 rs4295627 single nucleotide polymorphism (SNP) and the glioma risk has been studied previously, but these studies have yielded conflicting results. The aim of the present study is to analyze this association more vigorously, by means of a meta-analysis. A comprehensive literature search was performed in databases PubMed and EMBASE. Six articles including 12 case-control studies in English with 11,368 controls and 5891 cases were eligible for the meta-analysis. We conducted subgroup analyses by the source of controls, ethnicity, and country. Our meta-analysis revealed that the rs4295627 SNP was associated with the glioma risk in a heterozygote model (TG versus TT: odds ratio = 1.35, 95% confidence interval = 1.26-1.45, P = 0.066). Moreover, our results suggested that the rs4295627 SNP was associated with a notably increased risk of glioma among Caucasians except for Swedes in 4 models (the homozygote model, recessive model, dominant model, and additive model). Nonetheless, in Sweden and China, the results showed no associations. No evidence of the publication bias was uncovered. Thus, our meta-analysis suggests that the rs4295627 SNP is associated with an increased risk of glioma. Additional studies are needed to derive more precise conclusion.
Assuntos
Neoplasias Encefálicas/genética , Glioma/genética , Peptídeos e Proteínas de Sinalização Intracelular/genética , Polimorfismo de Nucleotídeo Único , Neoplasias Encefálicas/etnologia , Estudos de Casos e Controles , China , Glioma/etnologia , Humanos , RNA Longo não Codificante , SuéciaRESUMO
Radish floral bud abortion (FBA) is an adverse biological phenomenon that occurs during reproduction. Although FBA occurs frequently, its mechanism remains unknown. To elucidate the molecular mechanism underlying FBA, we detected gene expression differences between aborted and normal buds of radish using cDNA-amplified fragment length polymorphism (AFLP) and real-time polymerase chain reaction (real-time PCR). A total of 221 differentially expressed transcript-derived fragments (TDFs) were detected by 256 cDNA-AFLP primer combinations, of which 114 were upregulated and 107 were downregulated in the aborted buds. A total of 54 TDFs were cloned and sequenced. A BLAST search revealed that all TDFs have homologous sequences and 29 of these corresponded to known genes, whose functions were mainly related to metabolism, stimulus response, transcriptional regulation, and transportation. Expressions of 6 TDFs with different functions were further analyzed by real-time PCR yielding expression profiling results consistent with the cDNA-AFLP analysis. Our results indicated that radish FBA is related to abnormalities in various physiological and biochemical plant processes.