RESUMEN
Procalcitonin (PCT) is a current, frequently-used marker for severe bacterial infection. The aim of this study was to develop a cost-effective detection kit for rapid quantitative and on-site detection of PCT. To develop the new PCT quantitative detecting kit, a double-antibody sandwich immunofluorescent assay was employed based on time-resolved immunofluorescent assay (TRFIA) combined with lateral flow immunoassay (LFIA). The performance of the new developed kit was evaluated in the aspects of linearity, precision, accuracy, and specificity. Two-hundred thirty-four serum samples were enrolled to carry out the comparison test. The new PCT quantitative detecting kit exhibited a higher sensitivity (0.08 ng/mL). The inter-assay coefficient of variation (CV) and the intra-assay CV were 5.4%-7.7% and 5.7%-13.4%, respectively. The recovery rates ranged from 93% to 105%. Furthermore, a high correlation (n = 234, r = 0.977, p < 0.0001) and consistency (Kappa = 0.875) were obtained when compared with the PCT kit from Roche Elecsys BRAHMS. Thus, the new quantitative method for detecting PCT has been successfully established. The results indicated that the newly-developed system based on TRFIA combined with LFIA was suitable for rapid and on-site detection for PCT, which might be a useful platform for other biomarkers in point-of-care tests.
Asunto(s)
Cromatografía de Afinidad , Biomarcadores , Calcitonina , Péptido Relacionado con Gen de Calcitonina , Sistemas de Atención de Punto , Precursores de ProteínasRESUMEN
OBJEVTIVE: To investigate the effects of Cyclocarya paliurus (C. paliurus) polysaccharides on the spleen injury of diabetic rats. METHODS: Animals were divided into 6 groups, including normal group, model group, control group, low-dose group of C. paliurus polysaccharides treatment, middle-dose group of C. paliurus polysaccharides treatment and high-dose group of C. paliurus polysaccharides treatment. Histological analysis of spleen was analyzed using hematoxilin and eosin. Levels of biological parameters and anti-oxidative enzymes were determined by spectrophotometry. Interleukin-7 (IL-7) and IL-10 were measured by enzyme-linked immunosorbent assay. RESULTS: Compared with that of model group, superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase level increased 78.63% (P < 0.05), 51.76% (P < 0.05), 2.95 times (P < 0.01) and 41.11% (P < 0.05) in the high-dose group of C. paliurus polysaccharides treatment, respectively. IL-7 and IL-10 increase 1.66 (P < 0.01) and 1.21 times (P < 0.01) in the high-dose group of C. paliurus polysaccharides treatment, respectively. CONCLUSION: It is suggested that C. paliurus polysaccharides may play a protecting role for spleen injury of diabetic rats by enhancing the antioxidative ability and evaluating the immunity.
Asunto(s)
Diabetes Mellitus Experimental , Juglandaceae , Animales , Antioxidantes/farmacología , Diabetes Mellitus Experimental/tratamiento farmacológico , Hojas de la Planta , Polisacáridos/uso terapéutico , Ratas , BazoRESUMEN
BACKGROUND: YTH domain family (YTHDF) 2 acts as a "reader" protein for RNA methylation, which is important in tumor regulation. However, the effect of YTHDF2 in liver hepatocellular carcinoma (LIHC) has yet to be elucidated. METHODS: We explored the role of YTHDF2 in LIHC based on publicly available datasets [The Cancer Genome Atlas (TCGA), International Cancer Genome Consortium (ICGC), and Gene Expression Omnibus (GEO)]. A bioinformatics approach was employed to analyze YTHDF2. Logistic regression analyses were applied to analyze the correlation between YTHDF2 expression and clinical characteristics. To evaluate the effect of YTHDF2 on the prognosis of LIHC patients, we used Kaplan-Meier (K-M) curves. Gene set enrichment analysis (GSEA) was undertaken using TCGA dataset. Univariate and multivariate Cox analyses were used to ascertain the correlations between YTHDF2 expression and clinicopathologic characteristics with survival. Genes co-expressed with YTHDF2 were identified and detected using publicly available datasets [LinkedOmics, University of California, Santa Cruz (UCSC), Gene Expression Profiling Interactive Analysis (GEPIA), and GEO]. Correlations between YTHDF2 and infiltration of immune cells were investigated by Tumor Immune Estimation Resource (TIMER) and GEPIA. RESULTS: mRNA and protein expression of YTHDF2 was significantly higher in LIHC tissues than in non-cancerous tissues. High YTHDF2 expression in LIHC was associated with poor prognostic clinical factors (high stage, grade, and T classification). K-M analyses indicated that high YTHDF2 expression was correlated with an unfavorable prognosis. Univariate and multivariate Cox analyses revealed that YTHDF2 was an independent factor for a poor prognosis in LIHC patients. GSEA revealed that the high-expression phenotype of YTHDF2 was consistent with the molecular pathways implicated in LIHC carcinogenesis. Analyses of receiver operating characteristic curves showed that YTHDF2 might have a diagnostic value in LIHC patients. YTHDF2 expression was associated positively with SF3A3 expression, which implied that they may cooperate in LIHC progression. YTHDF2 expression was associated with infiltration of immune cells and their marker genes. YTHDF2 had the potential to regulate polarization of tumor-associated macrophages, induce T-cell exhaustion, and activate T-regulatory cells. CONCLUSION: YTHDF2 may be a promising biomarker for the diagnosis and prognosis of LIHC and may provide new directions and strategies for LIHC treatment.
RESUMEN
BACKGROUND: Increasing evidence suggests that aberrant alternative splicing (AS) events are associated with progression of cancer. This study evaluated the prognostic value and clarify the role of AS events in cervical cancer (CC). METHODS: Based on RNA-seq AS event data and clinical information of CC patients in The Cancer Genome Atlas (TCGA) database, we sought to identify prognosis-related AS events in this setting. We selected several survival-associated AS events to construct a prognostic predictor for CC through the least absolute shrinkage and selection operator (LASSO) and multivariate Cox regression. Moreover, Kyoto Encyclopedia of Genes and Genomes and Gene Ontology analyses were performed on genes with prognosis-related AS events and constructed an AS-splicing factors (SFs) regulatory network. RESULTS: 2770 AS events were significantly correlated with overall survival (OS). The area under the curve (AUC) values of receiver-operator characteristic curve (ROC) for the final prognostic predictor were 0.926, 0.946 and 0.902 at 3, 5, and 10 years, respectively. These values indicated efficiency in prognostic risk stratification for patients with CC. The final prognostic predictor was an independent predictor of OS (HR: 1.24; 95% CI: 1.020-1.504; P < 0.05). The AS-SFs correlation network may reveal an underlying regulatory mechanism of AS events. CONCLUSION: AS events are essential participants in the prognosis of CC and hold great potentials for the prognostic stratification and development of treatment strategy.
RESUMEN
BACKGROUND: The aim of the present study was to investigate the diagnostic value of glypican-3 (GPC3), arginase-1 (Arg-1), and hepatocyte paraffin antigen 1 (HepPar-1) in differentiating hepatocellular carcinoma (HCC) from intrahepatic cholangiocarcinoma (ICC). METHODS: The expression of GPC3, HepPar-1 and Arg-1 were measured by immunohistochemistry in 47 cases of HCC, 29 cases of ICC and their paracancerous tissues. RESULTS: A high expression of GPC3, Arg-1 and HepPar-1 was observed in HCC tissues (68.09%, 76.60% and 78.72%, respectively; P>0.0125) while it was lower in ICC tissues (6.90%, 6.90% and 13.79%, respectively; P>0.0125). With regard to specificity, GPC3 performed better than Arg-1 and HepPar-1 (97.37% vs. 1.32% and 2.63%, respectively; P<0.05). The positive rate in poorly differentiated HCC for either GPC3, Arg-1 or HepPar-1 was lower than that in well- and moderately differentiated HCC. The majority of positive samples for GPC3 and Arg-1 were grade 2+ in well-, moderately- or poorly-differentiated HCC. Combined detection of GPC3, Arg-1 and HepPar-1 could increase the sensitivity up to 89.36% and the specificity to 100.00%, comparing with any single biomarker (P<0.05). CONCLUSIONS: GPC3, Arg-1 and HepPar-1 were all useful biomarkers in differentiating HCC from ICC. The combination models could improve the diagnosis value of HCC and help differentiating HCC from ICC.
RESUMEN
OBJEVTIVE: To investigate the efficacy of Cyclocarya paliurus (C. paliurus) polysaccharides on stre- ptozotocin-induced diabetic nephropathy in rats. METHODS: Rats were divided into 6 groups, including group of normal control, group of diabetic control, group of metformin treatment, low-dose group of C. paliurus polysaccharides treatment, middle-dose group of C. paliurus polysaccharides treatment and high-dose group of C. paliurus polysaccharides treatment. Histological analysis of kidney was analyzed using hematoxilin and eosin. Levels of blood glucose, creatinine, urea, uric acid were determined by spectrophotometry. Anti-oxidative enzymes were measured by real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). Advanced glycation end products (AGEs) and transforming growth factor-ß1 (TGF-ß1) level was measured by ELISA. RESULTS: Abnormal changes were observed in the group of diabetic control characterized by atrophy of the renal glomeruli with hypercellularity, congestion of glomerular tufts, dilation of the renal spaces, and degeneration of renal tubule. Compared with that of normal group, blood glucose, creatinine, urea, uric acid level was significantly increased in the group of diabetic control. Superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase level was significantly decreased, but AGEs and TGF-ß1 level was significantly increased. By contrast, administration of C. paliurus polysaccharides and metformin could reverse the above-mentioned results of the group of diabetic control, especially in the high-dose group of C. paliurus polysaccharides. CONCLUSION: Our findings suggest that C. paliurus polysaccharides may play a protecting role for nephropathy of diabetic rats by lowering glucose, creatinine, urea, uric acid level, enhancing the antioxidative ability, and reducing AGEs and TGF-ß1 expression.
Asunto(s)
Nefropatías Diabéticas/prevención & control , Medicamentos Herbarios Chinos/administración & dosificación , Juglandaceae/química , Polisacáridos/administración & dosificación , Sustancias Protectoras/administración & dosificación , Animales , Glucemia/metabolismo , Nefropatías Diabéticas/metabolismo , Glutatión Peroxidasa/metabolismo , Humanos , Riñón/efectos de los fármacos , Riñón/metabolismo , Masculino , Estrés Oxidativo/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Estreptozocina , Superóxido Dismutasa/metabolismoRESUMEN
Inflammation and immunity are important determinants of cancer initiation, promotion, and progression to cancer equilibrium or suppression. The NOD-like receptor family pyrin domain containing 3 (NLRP3) is an oligomeric intracellular immune receptor, and the main component of inflammasome. As a widely distributed effector of innate immunity, NLRP3 inflammasome affects development of many cancer types, but its exact role in colorectal cancer (CRC) is controversial. We found that cells with the macrophage (MΦ) marker CD68 and strong NLRP3 expression densely surrounded CRC tissue. The NLRP3 inflammasome was activated in MΦs by MΦ-CRC cell crosstalk; it resulted in faster migration of CRC cells, whereas blocking NLRP3 signaling suppressed CRC cell migration in vitro, and metastatic ability in vivo. NLRP3 signaling activation in MΦs can contribute to CRC cell migration and invasion.
Asunto(s)
Movimiento Celular , Neoplasias Colorrectales/metabolismo , Inflamasomas/metabolismo , Neoplasias Hepáticas/metabolismo , Macrófagos/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Comunicación Paracrina , Animales , Técnicas de Cocultivo , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , Medios de Cultivo Condicionados/metabolismo , Humanos , Interleucina-1beta/metabolismo , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/secundario , Macrófagos/patología , Masculino , Ratones , Ratones de la Cepa 129 , Ratones Endogámicos C57BL , Ratones Noqueados , Proteína con Dominio Pirina 3 de la Familia NLR/deficiencia , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Invasividad Neoplásica , Fenotipo , Transducción de Señal , Células THP-1RESUMEN
INTRODUCTION: Dexmedetomidine (DEX) has been demonstrated to inhibit inflammatory response and protect against multiorgan injury in various scenarios. The objectives of the present study were to ascertain whether DEX is able to attenuate acute lung injury (ALI) under heatstroke (HS), and to explore the underlying mechanism. METHODS: Male C57BL/6 mice were exposed to ambient temperature of 39.5â±â0.2°C until core temperature reach 43°C. DEX or 0.9% saline was injected i.p. immediately. At the end of the experiment, bronchoalveolar lavage fluid (BALF) and lung tissue were harvested. RESULTS: HS induce ALI and pulmonary dysfunction, while DEX treatment could significantly inhibit lung injury and improve respiratory dysfunction under HS. The overall effect was beneficial and improved the 72âh cumulative survival rate of mice with HS. Furthermore, HS significantly elevated the levels of cytokines in BALF, as well as increased the activity of toll-like receptor 4 (TLR4)/MyD88/nuclear factor-κB (NFκB) signaling pathway in lung tissue, while DEX treatment could inhibit such effects. Finally, DEX could upregulate the expression of caveolin 1 downregulated by HS, which may contribute to the inhibition of TLR4/MyD88/NFκB signaling pathway. DISCUSSION: In conclusion, the present results indicated that DEX may protect against lung inflammatory response and injury under HS via TLR4/MyD88/NFκB signaling pathway, and caveolin-1 may participate in the effects.
Asunto(s)
Lesión Pulmonar Aguda , Dexmedetomidina/farmacología , Trastornos de Estrés por Calor , Lesión Pulmonar Aguda/tratamiento farmacológico , Lesión Pulmonar Aguda/etiología , Lesión Pulmonar Aguda/metabolismo , Lesión Pulmonar Aguda/patología , Animales , Trastornos de Estrés por Calor/complicaciones , Trastornos de Estrés por Calor/tratamiento farmacológico , Trastornos de Estrés por Calor/metabolismo , Trastornos de Estrés por Calor/patología , Masculino , Ratones , Factor 88 de Diferenciación Mieloide/metabolismo , FN-kappa B/metabolismo , Transducción de Señal/efectos de los fármacos , Receptor Toll-Like 4/metabolismoRESUMEN
Recently, accumulating evidence from clinical and experimental researches have suggested that translationally controlled tumor protein (TCTP) and high mobility group box 1 (HMGB1) are implicated in colorectal cancer (CRC) metastasis. However, whether there is an interconnection between these two tumor-promoting proteins and how they affect CRC metastasis remain to be fully elucidated. In the present study, the expression level of TCTP in CRC tissues was assessed by immunohistochemical staining and immunoblotting, and the serum concentration of HMGB1 in patients with CRC was detected by enzyme-linked immunosorbent assay. In vitro, following the modulation of TCTP expression in colon cancer LoVo cells, the translocation behavior of HMGB1 was observed by immunofluorescence assay. Furthermore, the activity of nuclear factor-κB (NF-κB) in LoVo cells was evaluated by immunoblotting and luciferase assay, and the invasion ability of LoVo cells after different treatments was determined using cell invasion assay. In vivo, xenograft tumor model was established and the correlation of TCTP and HMGB1 expression in xenografted tumors was studied by immunohistochemical examination. The results revealed that the expression level of TCTP in CRC tissue and the serum concentration of HMGB1 in patients with CRC were significantly increased, and there was a strong positive correlation between them. In vitro experiments showed that the overexpression of TCTP on LoVo cells resulted in the release of HMGB1 from the nucleus to the cytoplasm and into the extracellular space. In addition, the overexpression of TCTP led to the activation of NF-κB in LoVo cells, and this effect was reversed by treatment with antibodies targeting HMGB1 or to its receptors Toll-like receptor 4 (TLR4) and receptor for advanced glycation end products advanced glycation end products (RAGE). Furthermore, inhibition of the HMGB1-TLR4/RAGE-NF-κB pathway significantly inhibited the TCTP-stimulated invasion of LoVo cells. In vivo experiments demonstrated that the overexpression of TCTP in nude mice promoted the development and spread of xenografted tumors, and concurrently enhanced the expression of HMGB1 in tumor tissues. Collectively, these findings suggested that TCTP promotes CRC metastasis through regulating the behaviors of HMGB1 and the downstream activation of the NF-κB signaling pathway.