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OBJECTIVE: Pancreatic ductal adenocarcinoma (PDAC) is the most lethal malignancy and lacks effective treatment. We aimed to understand molecular mechanisms of the intertwined interactions between tumour stromal components in metastasis and to provide a new paradigm for PDAC therapy. DESIGN: Two unselected cohorts of 154 and 20 patients with PDAC were subjected to correlation between interleukin (IL)-33 and CXCL3 levels and survivals. Unbiased expression profiling, and genetic and pharmacological gain-of-function and loss-of-function approaches were employed to identify molecular signalling in tumour-associated macrophages (TAMs) and myofibroblastic cancer-associated fibroblasts (myoCAFs). The role of the IL-33-ST2-CXCL3-CXCR2 axis in PDAC metastasis was evaluated in three clinically relevant mouse PDAC models. RESULTS: IL-33 was specifically elevated in human PDACs and positively correlated with tumour inflammation in human patients with PDAC. CXCL3 was highly upregulated in IL-33-stimulated macrophages that were the primary source of CXCL3. CXCL3 was correlated with poor survival in human patients with PDAC. Mechanistically, activation of the IL-33-ST2-MYC pathway attributed to high CXCL3 production. The highest level of CXCL3 was found in PDAC relative to other cancer types and its receptor CXCR2 was almost exclusively expressed in CAFs. Activation of CXCR2 by CXCL3 induced a CAF-to-myoCAF transition and α-smooth muscle actin (α-SMA) was uniquely upregulated by the CXCL3-CXCR2 signalling. Type III collagen was identified as the CXCL3-CXCR2-targeted adhesive molecule responsible for myoCAF-driven PDAC metastasis. CONCLUSIONS: Our work provides novel mechanistic insights into understanding PDAC metastasis by the TAM-CAF interaction and targeting each of these signalling components would provide an attractive and new paradigm for treating pancreatic cancer.
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Fibroblastos Associados a Câncer/metabolismo , Carcinoma Ductal Pancreático/patologia , Quimiocinas CXC/metabolismo , Neoplasias Pancreáticas/patologia , Macrófagos Associados a Tumor/metabolismo , Animais , Carcinoma Ductal Pancreático/mortalidade , Estudos de Coortes , Humanos , Interleucina-33/metabolismo , Camundongos Knockout , Metástase Neoplásica , Neoplasias Pancreáticas/mortalidade , Regulação para CimaRESUMO
Zinc bioavailability with the presence of other elements in wheat grains might be affected by fertilizers. A long-term field experiment was conducted to examine effects of N fertilizer on Zn bioavailability in wheat grain tissues, with changes in the concentrations, distribution, and speciation of Zn as well as P and sulfur S via synchrotron-based technology. Results showed that addition of N fertilizer was associated with changes in Zn concentrations and distributions in grain tissues, especially in the crease region and endosperm. Simultaneously, N addition enhanced Zn-S colocalization in the crease region and endosperm and lowered the P/Zn ratio and Zn-P colocalization. Addition of N fertilizer with P increased Zn-cysteine (9.2%) and decreased Zn-phytate (47.3%) in the crease region, leading to potentially higher grain Zn bioavailability. Thus, addition of N fertilizer improved concentrations and bioavailability of Zn, by coordinating the relationships among Zn, P and S within wheat grains.
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Fertilizantes , Triticum , Fertilizantes/análise , Disponibilidade Biológica , Grão Comestível/química , ZincoRESUMO
Near-infrared (NIR) fluorescence optical imaging is an emerging imaging technique for studying diseases at the molecular level. Optical imaging with a NIR emitting fluorophore for targeting tumor vasculature offers a noninvasive method for early detection of tumor angiogenesis and efficient monitoring of response to anti-tumor vasculature therapy. The previous in vitro results demonstrated that the GX1 peptide, identified by phage-display technology, is a tumor vasculature endothelium-specific ligand. In this report, Cy5.5-conjugated GX1 peptide was evaluated in a subcutaneous U87MG glioblastoma xenograft model to investigate tumor-targeting efficacy. The in vitro flow cytometry results revealed dose-dependent binding of Cy5.5-GX1 peptide to U87MG glioma cells. In vivo optical imaging with the Cy5.5-GX1 probe exhibited rapid U87MG tumor targeting at 0.5 h p.i., and high tumor-to-background contrast at 4 h p.i. Tumor specificity of Cy5.5-GX1 was confirmed by effective blocking of tumor uptake in the presence of unlabeled GX1 peptide (20 mg/kg). Ex vivo imaging further confirmed in vivo imaging findings, and demonstrated that Cy5.5-GX1 has a tumor-to-muscle ratio (15.21 ± 0.84) at 24 h p.i. for the non-blocked group and significantly decreased ratio (6.95 ± 0.75) for the blocked group. In conclusion, our studies suggest that Cy5.5-GX1 is a promising molecular probe for optical imaging of tumor vasculature.
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Carbocianinas , Imagem Molecular/métodos , Sondas Moleculares , Biblioteca de Peptídeos , Peptídeos , Neoplasias Gástricas/diagnóstico , Animais , Bacteriófagos/química , Bacteriófagos/genética , Bacteriófagos/metabolismo , Carbocianinas/química , Linhagem Celular Tumoral , Feminino , Corantes Fluorescentes/química , Humanos , Camundongos , Camundongos Nus , Imagem Molecular/instrumentação , Sondas Moleculares/química , Sondas Moleculares/genética , Neovascularização Patológica , Peptídeos/química , Peptídeos/genética , Espectrometria de Fluorescência/métodos , Espectroscopia de Luz Próxima ao Infravermelho/métodos , Estômago/irrigação sanguínea , Estômago/química , Neoplasias Gástricas/químicaRESUMO
Plastic mulching (PM) is widely used to improve crop water use efficiency and grain yield, but few studies have reported the effects of PM on cereal crop quality, especially sulfur (S) nutrition of wheat, which has significant effects on grain protein content, dough rheology, baking quality and human health. To fill this knowledge gap, we conducted a multi-site field experiment on the Loess Plateau from 2014 to 2016 to study the effects of PM combined with nitrogen (N) fertilizer on grain yield, shoot S accumulation, and grain S concentration of winter wheat in dryland. Compared with no mulching (NM), PM increased grain yield by 13.7% but decreased grain S concentration, S requirement for 1,000 kg-1 grain, soil available S concentration, and post-anthesis S uptake by 9.0, 9.7, 24.4, and 51.8%, respectively. Plastic mulching significantly increased shoot S accumulation at anthesis by 19.2%, but there was no significant difference at maturity. Additionally, grain S concentration and S requirement had a linear-plateau relationship with N fertilization amount, reaching maximum values at 110 and 127 kg N ha-1 under PM, 37.5 and 27.0% higher than those under NM. Furthermore, shoot S accumulation and N application rates well-fitted the linear-plateau model at anthesis and maturity. At maturity, straw, grain, and shoots accumulated the most S at threshold N rates of 120, 85 and 110 kg N ha-1, respectively. Crucially, stem + leaf S concentration at anthesis had a significant linear relationship with grain S concentration under PM; a 1 g kg-1 increase in stem leaf concentration corresponded with a 0.24 g kg-1 increase in grain S concentration. This study's findings suggest that combining soil S supplementation with optimal N fertilizer under PM in northwest China and other regions with similar cropping systems increases grain S concentration and improves nutritional and processing qualities.
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Increasing iron (Fe) and zinc (Zn) concentrations in crop grains with high yield is an effective measure to ensure food supply and alleviate mineral malnutrition in humans. Micronutrient concentrations in grains depend on not only their availability in soils but also their uptake in roots and translocation to shoots and grains. In this three-year field study, we investigated genotypic variation in Fe and Zn uptake and translocation within six wheat cultivars and examined in detail Fe and Zn distributions in various tissues of two cultivars with similar high yield but different grain Fe and Zn concentrations using synchrotron micro-X-ray fluorescence. Results revealed that root Fe and Zn concentrations were 11 and 44% greater in high-nutrient (HN) than in low-nutrient (LN) concentration cultivar. Although both cultivars accumulated similar amounts of Fe in shoots, HN cultivar had greater accumulation of Fe in grain and greater accumulation of Zn in both shoots and grain. Grain Zn concentration was positively correlated with shoot Zn accumulation, and grain Fe concentration was positively correlated with the ability to translocate Fe from leaves/stem to grains. In the first nodes of shoots, HN cultivar had 482% greater Fe and 36% greater Zn concentrations in the enlarged vascular bundle (EVB) than LN cultivar. In top nodes, HN cultivar had 225 and 116% greater Fe and Zn concentrations in the transit vascular bundle and 77 and 71% greater in the EVB when compared to LN cultivar. HN cultivar also had a greater ability to allocate Fe and Zn to the grain than LN cultivar. In conclusion, HN cultivar had greater capacity of Fe and Zn acquirement by roots and translocation and partitioning from shoots into grains. Screening wheat cultivars for larger Fe and Zn concentrations in shoot nodes could be a novel strategy for breeding crops with greater grain Fe and Zn concentrations.
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Triticum , Zinco , Grão Comestível , Fluorescência , Humanos , Ferro , Melhoramento Vegetal , Síncrotrons , Triticum/genética , Raios XRESUMO
The physiological disorders in humans resulting from the excess dietary intake of manganese (Mn) via whole-grain food has attracted considerable attention. However, the speciation and bioavailability of Mn in wheat grains and their response to different phosphorus (P) fertilization rates are still unclear. In the current study, using a long-term field trial with P application rates of 0, 21.8, 43.6, 65.5 and 87.3 kg/ha, we examined changes in the concentration, distribution, and speciation of Mn of wheat grains using synchrotron-based X-ray fluorescence microscopy and X-ray absorption spectroscopy. The total Mn concentration in grains was found to be increased by phosphorus fertilization, especially in embryo in the form of Mn(II), but this phosphorus fertilization also decreased Mn concentrations in the nucellar projection. In this study, the speciation of Mn in different wheat grain tissues was examined, and results indicate that in calcareous soils, high rates of P fertilizers can increase Mn concentrations in wheat grain, including Mn which is likely to be of high bioavailability, and thus may increase the risk for human to expose to high Mn intake via whole-grain food.
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Solo , Triticum , Fertilização , Fertilizantes/análise , Humanos , Manganês , FósforoRESUMO
Improving the concentration and bioavailability of zinc (Zn) in cereal grains is an important way to solve the problem of Zn deficiency in human body. The bioavailability of Zn is related to both its distribution and speciation in grains. In the current study, we examined the differences of Zn concentration, distribution, and speciation within grains among wheat cultivars with similar high grain yield but contrasting grain Zn concentration using synchrotron micro X-ray fluorescence (µ-XRF) and X-ray absorption near-edge structure (XANES). Results showed that compared to the low-Zn cultivar, the Zn concentration was 103, 50, 76, 33, and 64% higher in the crease region, aleurone layer, scutellum, embryonic axis, and endosperm of the high-Zn cultivar, respectively. Zinc mainly colocalized with phosphorus (P) in the aleurone layer and the scutellum, but less colocalization of Zn with P and a much lower concentration ratio of P/Zn were found in the high-Zn cultivar. Sulfur (S) is present in the form of scattered spots in the endosperm in accord with Zn, but the colocalization of Zn with S was predominant in the modified aleurone layer and the nucellar projection of the high-Zn cultivar. XANES results showed the lower proportion of Zn-phytate in the high-Zn cultivar. Findings indicated that it is possible to select the high-yield wheat cultivar with both high grain Zn concentration and high bioavailability, which provide a new perspective for genetic Zn biofortification.
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Grão Comestível , Triticum , Biofortificação , Disponibilidade Biológica , Humanos , ZincoRESUMO
PURPOSE: To synthesize the dimer of GX1 and identify whether its affinity and targeting are better than those of GX1. To prepare 68Ga-DOTA-KEK-(GX1)2 and to apply it to PET and Cerenkov imaging of gastric cancer. METHODS: 68Ga-DOTA-KEK-(GX1)2 was prepared, and the labeling yield and stability were determined. Its specificity and affinity were verified using an in vitro cell binding assay and competitive inhibition test, cell immunofluorescence, and cell uptake and efflux study. Its tumor-targeting ability was determined by nano PET/CT and Cerenkov imaging, standardized uptake value (SUV), signal-to-background ratio (SBR) quantification, and a biodistribution study in tumor-bearing nude mice. RESULTS: 68Ga-DOTA-KEK-(GX1)2 was successfully prepared, and the labeling yield was more than 97%. It existed stably for 90 min in serum. The binding of 68Ga-DOTA-KEK-(GX1)2 to cocultured HUVECs (Co-HUVECs) was higher than that to human umbilical vein endothelial cells (HUVECs), BGC823 cells, and GES cells. It was also higher than that of 68Ga-DOTA-GX1, indicating that the dimer did improve the specificity and affinity of GX1. The binding of KEK-(GX1)2 to Co-HUVECs was significantly higher than that of GX1. Additionally, the uptake of 68Ga-DOTA-KEK-(GX1)2 by Co-HUVECs was higher than that of 68Ga-DOTA-GX1 and reached a maximum at 60 min. Nano PET/CT and Cerenkov imaging showed that the tumor imaging of the nude mice injected with 68Ga-DOTA-KEK-(GX1)2 was clear, and the SUV and SBR value of the tumor sites were significantly higher than those of the nude mice injected with 68Ga-DOTA-GX1, indicating that the probe had better targeting in vivo. Finally, the biodistribution showed quantitatively that when organs such as the kidney and liver metabolized rapidly, the radioactivity of the tumor site of the nude mice injected with 68Ga-DOTA-KEK-(GX1)2 decreased relatively slowly. At the same time, the percentage of injected dose per gram (%ID/g) of the tumor site was higher than that of other normal organs except the liver and kidney at 60 min, which indicated that the tumor had good absorption of the probe. CONCLUSION: GX1 was modified successfully, and the in vivo and in vitro properties of the GX1 dimer were significantly better than those of GX1. The imaging probe, 68Ga-DOTA-KEK-(GX1)2, was successfully prepared, which provides a candidate probe for PET and Cerenkov diagnosis of gastric cancer.
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Identification of molecules specific to the retinal neovasculature will promote antiangiogenic therapy with enhanced targeting ability. The specificity of phage-displayed peptide GX1 (a cyclic 7-mer peptide motif CGNSNPKSC) to gastric cancer neovasculature has been extensively confirmed both in vitro and in vivo. To investigate the potential application of GX1 in antiangiogenic therapy targeting retinal angiogenesis-related diseases, we performed immunohistochemistry and immunofluorescence analyses. GX1 demonstrated positive staining in the retinal neovasculature in an oxygen-induced mouse model of retinopathy (OIR) as well as in rat retinal microvasculature endothelial cells (RMECs), confirming the major role of the GX1 receptor during retinal angiogenesis. Dimeric GX1 was synthesized to increase the binding affinity to the GX1 receptor, and the antiangiogenic effects were examined in RMECs in vitro and the retinal neovasculature in the OIR in vivo. Cell proliferation was evaluated using a Cell Counting Kit-8 (CCK-8) assay, revealing that compared with the GX1 monomer, dimeric GX1 significantly inhibited RMEC proliferation (P < 0.05). This finding may be attributed to the enhanced (P < 0.05) apoptosis induced by dimeric GX1 in RMECs based on results obtained from TUNEL, flow cytometric and cell cycle analyses. In RMECs, in vitro cell migration and tube formation were significantly inhibited following exposure to dimeric GX1. Intravitreal administration of dimeric GX1 resulted in a greater reduction in the retinal neovascularization in vivo than administration of the GX1 monomer (P < 0.05). In conclusion, dimeric GX1 showed greater inhibition of angiogenesis than monomeric GX1 and could be a promising agent for antiangiogenic therapy in retinal angiogenesis-related diseases.
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Inibidores da Angiogênese/farmacologia , Neovascularização Patológica/tratamento farmacológico , Peptídeos/farmacologia , Neovascularização Retiniana/tratamento farmacológico , Inibidores da Angiogênese/uso terapêutico , Animais , Apoptose/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Dimerização , Modelos Animais de Doenças , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/patologia , Camundongos Endogâmicos C57BL , Peptídeos/uso terapêutico , Ratos , Neovascularização Retiniana/patologiaRESUMO
PURPOSE: The GX1 peptide (CGNSNPKSC) can specifically bind to TGM2 and possesses the ability to target the blood vessels of gastric cancer. This study intends to develop an integrated dual-functional probe with higher affinity, specificity and targeting and to characterize it in vivo and in vitro. METHODS: The dimer and tetramer of GX1 were prepared using cross-linked PEG and labeled with 99mTc. The best targeting probe [PEG-(GX1)2] was selected by gamma camera imaging in nude mouse models of gastric cancer. 188Re-PEG-(GX1)2 was prepared and characterized through cell binding analysis and competitive inhibition experiments, gamma camera imaging, MTT analysis and flow cytometry, BLI, immunohistochemistry, HE staining and biochemical analysis. RESULTS: PEG-(GX1)2 bound specifically to Co-HUVEC with higher affinity than GX1. 188Re-PEG-(GX1)2 had better ability to target gastric cancer in tumor-bearing nude mice and higher T/H ratios than 188Re-GX1. 188Re-PEG-(GX1)2 inhibited the growth of Co-HUVEC and induced apoptosis, and its effects were more robust than those of 188Re-GX1. BLI showed that 188Re-PEG-(GX1)2 inhibited tumor proliferation in vivo with a stronger effect than 188Re-GX1. Compared with 188Re-GX1, 188Re-PEG-(GX1)2 suppressed tumor angiogenesis and tumor cell proliferation and induced tumor cell apoptosis in vivo. The 188Re-PEG-(GX1)2 group did not cause visible changes in liver and kidney morphology and function in vivo. CONCLUSION: The dimer of GX1 was synthesized by using cross-linked PEG, and then 188Re-PEG-(GX1)2 was prepared. This radiopharmaceutical played both diagnostic and therapeutic functions, and gamma camera imaging could be utilized to detect the distribution of drugs in vivo during treatment. Through a series of experiments in vitro and in vivo, the feasibility of the drug was confirmed, and these results laid the foundation for the subsequent development and application of GX1.
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Inibidores da Angiogênese/metabolismo , Proteínas de Ligação ao GTP/metabolismo , Imagem Molecular/métodos , Fragmentos de Peptídeos/metabolismo , Radioisótopos/metabolismo , Rênio/metabolismo , Neoplasias Gástricas/metabolismo , Transglutaminases/metabolismo , Inibidores da Angiogênese/uso terapêutico , Animais , Linhagem Celular Tumoral , Feminino , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Sondas Moleculares/metabolismo , Proteína 2 Glutamina gama-Glutamiltransferase , Neoplasias Gástricas/tratamento farmacológico , Neoplasias Gástricas/patologia , Carga Tumoral/efeitos dos fármacos , Carga Tumoral/fisiologiaRESUMO
BACKGROUND: The discovery of the importance of angiogenesis in tumor growth has emphasized the need to find specific vascular targets for tumor-targeted therapies. Previously, using phage display technology, we identified the peptide GX1 as having the ability to target the gastric cancer vasculature. The present study investigated the bioactivities of GX1, as well as its potential ability to cooperate with recombinant mutant human tumor necrosis factor alpha (rmhTNFalpha), in gastric cancer therapy. RESULTS: Tetrazolium salt (MTT) assay showed that GX1 could inhibit cell proliferation of both human umbilical vein endothelial cells (HUVEC) (44%) and HUVEC with tumor endothelium characteristics, generated by culturing in tumor-conditioned medium (co-HUVEC) (62%). Flow-cytometry (FCM) and western blot assays showed that GX1 increased the rate of apoptosis from 11% to 31% (p < 0.01) by up-regulating caspase 3 expression level. A chorioallantoic membrane assay indicated that GX1 could suppress neovascularization in vivo, with the microvessel count decreasing from 21 to 11 (p < 0.05). When GX1 was fused to rmhTNFalpha, GX1-rmhTNFalpha selectively concentrated in the gastric cancer vasculature, as shown by enzyme-linked immunosorbent assay, immunofluorescence and emission-computed tomography. In vitro MTT and FCM assays showed that, compared to rmhTNFalpha alone, GX1-rmhTNFalpha was more effective at suppressing co-HUVEC proliferation (45% vs. 61%, p < 0.05) and inducing apoptosis (11% vs. 23%, p < 0.05). In a tumor formation test, GX1-rmhTNFalpha more effectively inhibited tumor growth than rmhTNFalpha (tumor volume: 271 mm3 vs. 134 mm3, p < 0.05), with less systemic toxicity as measured by body weight (20.57 g vs. 19.30 g, p < 0.05). These therapeutic effects may be mediated by selectively enhanced tumor vascular permeability, as indicated by Evan's blue assay. CONCLUSION: GX1 had both homing activity and the ability to inhibit vascular endothelial cell proliferation in vitro and neovascularization in vivo. Furthermore, when GX1 was conjugated to rmhTNFalpha, the fusion protein was selectively delivered to targeted tumor sites, significantly improving the anti-tumor activity of rmhTNFalpha and decreasing systemic toxicity. These results demonstrate the potential of GX1 as a homing peptide in vascular targeted therapy for gastric cancer.
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Inibidores da Angiogênese/uso terapêutico , Antineoplásicos/uso terapêutico , Oligopeptídeos/uso terapêutico , Neoplasias Gástricas/irrigação sanguínea , Neoplasias Gástricas/tratamento farmacológico , Fator de Necrose Tumoral alfa/uso terapêutico , Animais , Apoptose/efeitos dos fármacos , Células Cultivadas , Progressão da Doença , Células Endoteliais/citologia , Células Endoteliais/efeitos dos fármacos , Humanos , Camundongos , Camundongos Nus , Peptídeos Cíclicos , Neoplasias Gástricas/patologia , Fator de Necrose Tumoral alfa/química , Fator de Necrose Tumoral alfa/metabolismoRESUMO
PURPOSE: To seek a high sensitive and convenient method for early diagnosis of gastric cancer by testing MG7-Ag in serum of gastric cancer patients and some other control groups using a convenient ELISA method. EXPERIMENT DESIGN: The expression of serum MG7-Ag was detected in 116 preoperative gastric cancer patients, 63 postoperative gastric cancer patients, 78 precancerous lesion patients, 50 healthy blood donors and patients of other cancers by a convenient ELISA method. For comparison, serum CEA, CA 50, CA 19-9 and TAG-72 were also detected in preoperative gastric cancer patients. Meanwhile, the expression of MG7-Ag was detected by immunohistochemical analysis in the groups of patients with gastric cancer or precancerous lesion mentioned above. RESULTS: The positive rate of Mg7-Ag determined by ELISA was 83. 6% of preoperative gastric cancer patients, 54.8% of lung cancer patients, 45.5% of rectal cancer patients, 17.6% of colonic cancer patients, 14.2% of breast cancer patients, 47.6% of postoperative gastric cancer patients, 12.8% of precancerous lesions patients and 0% of healthy blood donors, respectively. The sensitivity of ELISA (83.6%) was found to be similar with that of immunohistochemistry (94%, p > 0. 01), while the false positive rate was lower (12.8% vs. 51.3%). MG7-Ag expression level in gastric cancer was correlated with tumor differentiation (p < 0. 01) and pathological stage (p < 0. 01). CONCLUSION: This ELISA method may be a non-invasive candidate method for screening of large population with high risk of gastric cancer.
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Antígenos de Neoplasias/sangue , Ensaio de Imunoadsorção Enzimática/métodos , Neoplasias Gástricas/sangue , Adulto , Idoso , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Sensibilidade e EspecificidadeRESUMO
Agricultural management methods, such as cultivation or fallowing, have led to significant changes in soil fertility and hence, crop yield. Such changes may have stemmed from changes in soil microbial communities and associated biogeochemical processes. This phenomenon is particularly true in organic-poor soil in the Loess Plateau of China. In this study, we examined three existing soil management regimes as part of a 10-year field experiment and evaluated their effects on fungal and bacterial community structures by performing high-throughput 454 pyrosequencing. These management regimes were (i) fertilized winter wheat (Triticum aestivum L.) (FW), (ii) continuous natural fallow with weeds but without crop grown (NF), and (iii) continuous bare fallow without weeds or crop grown (BF). After 10 years, soil organic carbon (SOC), microbial biomass carbon (MBC), and available potassium (K) concentrations were highest in NF. Soil N behaved differently, with BF obtaining the highest nitrate nitrogen (N). Meanwhile, slight differences in total N (TN) were observed among FW, NF, and BF. Available phosphorus (P) was highest and available K was lowest in FW. Microbial communities were dominated by Ascomycota (59.1% of fungal sequences), and Acidobacteria, Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria (75.7% of bacterial sequences) in FW, NF and BF at the phylum level. Soil management regimes did not affect the fungal and bacterial richness and diversity but significantly modified their community compositions. Compared with FW, the abundances of Ascomycota (fungi phylum) and Alternaria, Gibberella, and Emericella (fungi genus) were increased by NF, whereas the values of Chaetomium, Humicola, and Cryptococcus (fungi genus) were decreased by BF. The abundances of Verrucomicrobia (bacteria phylum), and Steroidobacter (bacteria genus) were increased by NF, and Bacteroides (bacteria genus) was increased by BF. Canonical correspondence analysis showed that SOC, available P, and TN might be the key factors in community formation. Therefore, the decadal absence of plants (BF) affected soil fertility by increased available K and nitrate N, whileas natural fallow (NF) affected soil fertility by increased SOC, available K, and MBC, and they all changed fungal and bacterial community compositions.
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Microbiologia do Solo , Solo/classificação , Triticum , Alternaria/isolamento & purificação , Ascomicetos/isolamento & purificação , Biomassa , Carbono , Chaetomium/isolamento & purificação , China , Cryptococcus/isolamento & purificação , Emericella/isolamento & purificação , Gibberella/isolamento & purificação , Nitrogênio/análise , Potássio/análiseRESUMO
PURPOSE: This study aimed to evaluate the potential of PEGylated dimeric GX1 peptide as a radiotracer for imaging of colorectal cancer vasculature in a LoVo tumor xenografted mouse model. PROCEDURES: The [(99m)Tc]PEG-(GX1)2 peptide was synthesized and identified. Confocal immunofluorescence analysis, receptor binding assay, and competitive inhibition assay were performed to evaluate the binding specificity and the receptor binding affinity of PEG-(GX1)2 to Co-human umbilical vein endothelial cells (HUVECs). Single photon emission computed tomography imaging and biodistribution were performed to evaluate the targeting ability of PEG-(GX1)2 to colorectal cancer. RESULTS: The studies in vitro suggested that PEG-(GX1)2 co-localized with Factor VIII in the perinuclear cytoplasm of Co-HUVECs and bound specifically to Co-HUVECs with a high affinity. The studies in vivo demonstrated that the targeting efficacy of PEG-(GX1)2 was superior to GX1. CONCLUSIONS: PEGylation improved the affinity and the targeting ability of the GX1 peptide. PEG-(GX1)2 is a more promising probe for imaging of colorectal vasculature than GX1.
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Neoplasias Colorretais/irrigação sanguínea , Neoplasias Colorretais/patologia , Imagem Molecular/métodos , Compostos Radiofarmacêuticos/farmacocinética , Tecnécio/farmacocinética , Animais , Linhagem Celular Tumoral , Humanos , Masculino , Camundongos , Camundongos Nus , Compostos Radiofarmacêuticos/química , Sensibilidade e Especificidade , Tecnécio/química , Distribuição TecidualRESUMO
Targeted radiopharmaceutical is an effective treatment for solid tumors. By labeling with radionuclides, targeting peptide could achieve both noninvasive diagnosis and targeted radionuclide therapy. In order to evaluate the potential applicability of GEBP11 peptides in diagnosis and radiotherapy of gastric cancer, in this study, iodine 131 labeled GEBP11 peptides, including a novel bifid PEGlylated GEBP11 trimer and its corresponding monomer, were developed. The clinical potential of GEBP11 peptides, such as tumor binding affinity and antitumor efficacy was demonstrated and assessed with multimodality imaging methods. Cerenkov and SPECT imaging showed higher tumor uptake for (131)I-2PEG-(GEBP11)3 (P<0.05, day 1; P<0.01, day 2; vs. monomer). Biodistribution studies indicated higher tumor accumulation and better T/NT of (131)I-2PEG-(GEBP11)3. Bioluminescence imaging exhibited a significant tumor growth suppression in (131)I-2PEG-(GEBP11)3 treated group (P<0.001 vs. control; P<0.01 vs. monomer). After treatment with (131)I-2PEG-(GEBP11)3, the tumor volume and vasculature decreased significantly, and the survival time was prolonged to 75.5days. Meanwhile, no significant hepatic or renal toxicity was observed with (131)I-2PEG-(GEBP11)3 administered. In conclusion, (131)I-2PEG-(GEBP11)3 could be a promising candidate for peptide-based targeting therapy of gastric cancer. 2PEG-(GEBP11)3 might be a potential drug delivery vehicle for the antiangiogenic therapy of gastric cancer.
Assuntos
Peptídeos/uso terapêutico , Neoplasias Gástricas/irrigação sanguínea , Neoplasias Gástricas/radioterapia , Estômago/irrigação sanguínea , Estômago/patologia , Animais , Células Endoteliais da Veia Umbilical Humana , Humanos , Radioisótopos do Iodo/química , Radioisótopos do Iodo/farmacocinética , Radioisótopos do Iodo/uso terapêutico , Camundongos Nus , Peptídeos/química , Peptídeos/farmacocinética , Compostos Radiofarmacêuticos/química , Compostos Radiofarmacêuticos/farmacocinética , Compostos Radiofarmacêuticos/uso terapêutico , Estômago/efeitos da radiação , Neoplasias Gástricas/diagnóstico , Neoplasias Gástricas/patologiaRESUMO
PURPOSE: Molecular imaging using positron emission tomography (PET) radiotracers targeted to tumor vasculature offers a noninvasive method for early detection of tumor angiogenesis and efficient monitoring of response to anti-tumor vasculature therapy. The previous in vitro results demonstrated that the GX1 peptide, identified by phage display technology, is a tumor vasculature endothelium-specific ligand. In this study, we evaluated a 64Cu-labeled GX1 peptide as a potential radiotracer for microPET imaging of tumor vasculature in a U87MG tumor xenografted mouse model. METHODS: Macrocyclic chelating agent 1,4,7,10-tetraazacyclododecane-N, N', N'', N'''-tetraacetic acid (DOTA)-conjugated GX1 peptide was synthesized and radiolabeled with 64Cu (t(1/2) = 12.7 h) in ammonium acetate buffer. The 64Cu-labeled GX1 peptide was then subjected to in vitro tumor cell uptake study, small animal PET and direct tissue sampling biodistribution studies in a U87MG tumor xenografted mouse model. RESULTS: The in vitro experiment demonstrated that 64Cu-DOTA-GX1 is stable in PBS with more than 91% of 64Cu-DOTA-GX1 peptide remaining intact after 24 h of incubation. Cellular uptake and retention studies revealed (64)Cu-DOTA-GX1 binds to U87MG glioma cells and has good tumor cell retention. For small animal PET imaging studies, the U87MG tumors were all clearly visible with high contrast to contralateral background at all measured time points after injection of 64Cu-DOTA-GX1 while high accumulation in liver and kidneys were also observed at early time points. The U87MG tumor uptake was determined to be the highest (7.97 ± 0.75%ID/g) at 24 h pi. The blocking experiment was achieved by co-injection of 64Cu-DOTA-GX1 with non-radiolabeled GX1 peptide (20 mg/kg) at 24 h pi, suggesting 64Cu-DOTA-GX1 is a target-specific tracer. Furthermore, the biodistribution results were consistent with the quantification of microPET imaging, demonstrating the highest ratio (16.09 ± 1.21) of tumor/muscle uptake of 64Cu-DOTA-GX1 at 24 h pi for non-blocking group and significant decreased ratio (6.57 ± 0.58) for blocking group. Finally, metabolic studies suggested that 64Cu-DOTA-GX1 is stable in mouse blood and urine in vivo at early time point while the metal transchelation may also occur in mouse liver and kidneys. CONCLUSION: Our studies demonstrate that 64Cu-DOTA-GX1 is a promising radiotracer for imaging tumor vasculature.
Assuntos
Radioisótopos de Cobre , Glioma/irrigação sanguínea , Glioma/diagnóstico por imagem , Peptídeos , Tomografia por Emissão de Pósitrons/métodos , Compostos Radiofarmacêuticos , Análise de Variância , Animais , Linhagem Celular Tumoral , Radioisótopos de Cobre/química , Radioisótopos de Cobre/farmacocinética , Estabilidade de Medicamentos , Feminino , Glioma/metabolismo , Humanos , Camundongos , Camundongos Nus , Imagem Molecular/métodos , Sondas Moleculares/química , Sondas Moleculares/farmacocinética , Neovascularização Patológica/diagnóstico por imagem , Biblioteca de Peptídeos , Peptídeos/química , Peptídeos/farmacocinética , Compostos Radiofarmacêuticos/química , Compostos Radiofarmacêuticos/farmacocinética , Distribuição Tecidual , Transplante HeterólogoRESUMO
Angiogenesis plays an essential role in tumor growth and metastasis and is a promising target for cancer therapy. We characterized the effects of selective CIAPIN1 inhibition on the angiogenesis gastric cancer cell line SGC7901 by stable transfection of CIAPIN1 siRNA. Our study has been shown that CIAPIN1 play the determined role in tumor growth and multidrug resistance. The conditioned media obtained from SGC7901 treated with CIAPIN1 siRNA suppressed in vitro the proliferation, migration and tube formation of human umbilical vein endothelial cells compared with untransfected cells or cells transfected with control vector alone. Furthermore, the stable transfection of CIAPIN1 siRNA inhibited in vivo tumorigenicity and angiogenesis. Our findings support that selective inhibition of CIAPIN1 alone plays an instrumental role on gastric cancer associated angiogenesis.
Assuntos
Peptídeos e Proteínas de Sinalização Intracelular/antagonistas & inibidores , Peptídeos e Proteínas de Sinalização Intracelular/genética , RNA Interferente Pequeno/genética , Neoplasias Gástricas/irrigação sanguínea , Neoplasias Gástricas/genética , Animais , Biomarcadores Tumorais/análise , Western Blotting , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Resistência a Múltiplos Medicamentos , Células Endoteliais/fisiologia , Regulação Neoplásica da Expressão Gênica , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Camundongos , Camundongos Nus , Neovascularização Patológica/genética , RNA Interferente Pequeno/metabolismo , Neoplasias Gástricas/metabolismo , Transfecção , Veias Umbilicais/fisiologiaRESUMO
Discovery of tumor vascular specific molecules to improve the targeting ability of cytotoxic agents plays an important role in antiangiogenesis. We had found a peptide GX1 (CGNSNPKSC) binding to vasculature endothelial cells of human gastric cancer by phage display technology and its specificity to vasculature had been thoroughly confirmed in vitro. To further evaluate the applicability of GX1 in antiangiogenesis therapy of gastric cancer, immunohistochemical analysis and ECT imaging in nude mice were performed. Immunohistochemical analysis showed that GX1 phage produced positive staining on 51/65 (78%) cases of the vasculature of gastric cancer. Simultaneously GX1 peptide was labeled with (99)Tc(m)O(4)(-), which obtained with high labeling efficiency. (99)Tc(m)-GX1 could specifically bind to Co-HUVEC and HUVEC with a binding constant of 3062 pM and 3831 pM respectively. ECT imaging indicated that GX1 could efficiently target to xenographic tissue in mice model with a high tumor/heart radio than that of control peptide. Biodistribution showed that tumor uptake was 0.74+/-0.02% ID/g at 24 h, 11 times than that of muscle. Immunofluorescence showed GX1 peptide could bind to xenograft vasculature in vivo. The results confirmed the targeting specificity of GX1 in gastric cancer-associated angiogenesis. It would be promising to further develop GX1 peptide-based assay for tumor angiogenesis imaging to improve diagnosis and internal radiotherapy.