RESUMO
Neuropilin-1 (NRP-1) is a nontyrosine kinase coreceptor for semaphorin 3A and the vascular endothelial growth factor involved in tumor angiogenesis, growth, and metastasis and is regarded as a promising target for cancer therapy. In the present study, we investigated the effects of an anti-NRP-1 monoclonal antibody (mAb) that we generated for MCF7 breast cancer cellular adhesion studies. MTT, colony formation, and adhesion assays showed that our anti-NRP-1 mAb dose-dependently inhibited MCF7 proliferation and fibronectin adhesion, leading to a rounded cellular morphology. Further, rhodamine phalloidin stain revealed that fibronectin-dependent formation of actin stress fibers was inhibited by anti-NRP-1 mAb. Immunoprecipitation and western blot showed that anti-NRP-1 mAb treatment inhibited the formation of NRP-1-α5ß1 integrin complexes and suppressed the phosphorylation of focal adhesion kinase and p130cas in MCF7 cells. These findings contribute to further understanding the NRP-1 function in cell adhesion and tumor metastasis. Moreover, our anti-NRP-1 mAb is a prospective drug candidate for tumor treatment.
Assuntos
Anticorpos Monoclonais/farmacologia , Neoplasias da Mama/patologia , Adesão Celular/efeitos dos fármacos , Proteína Substrato Associada a Crk/metabolismo , Fibronectinas/fisiologia , Proteína-Tirosina Quinases de Adesão Focal/metabolismo , Neuropilina-1/metabolismo , Actinas/metabolismo , Anticorpos Monoclonais/isolamento & purificação , Feminino , Humanos , Células MCF-7 , Neuropilina-1/imunologia , Transdução de SinaisRESUMO
OBJECTIVE: To explore the application of folding transfer shelf in the transportation of critically ill patients. METHODS: Patients transferred from the emergency department to the intensive care unit (ICU) admitted to the First Hospital of Jiaxing from January 1st to December 31st in 2019 were enrolled. The patients were divided into study group and control group by whether or not using the self-developed folding transfer shelf. The incidence of adverse events, the stability rate of vital signs and the transport time were compared between the two group. RESULTS: A total of 437 patients were enrolled in the study, with 222 in the study group (which used the self-developed folding transfer shelf) and 215 in the control group (which used the conventional stretcher). The baseline data such as gender, age, disease status and disease severity were balanced between the two groups. The stability rate of vital signs in the study group was higher than that in the control group (89.19% vs. 82.33%, P < 0.05). The transfer time in the study group was shorter than that in the control group (minutes: 6.39±1.35 vs. 7.61±1.34, P < 0.01). The total incidence of adverse transport events in the study group was lower than that in the control group (2.25% vs. 10.23%, P < 0.01). The incidence of miscarriage of emergent materials and instrument falling in the study group were lower than those in the control group (0% vs. 2.79%, 0% vs. 2.33%, both P < 0.05). CONCLUSIONS: The folding transfer shelf could reduce the transport risk of critical ill patients, especially the risk of miscarriage and falling of rescue instrument. The application of folding transfer shelf could regulate the management of transport, keep the vital signs of patients stable during transport, shorten the transport time, and facilitate an efficient and high-quality transport.
Assuntos
Estado Terminal , Cuidados Críticos , Serviço Hospitalar de Emergência , Humanos , Unidades de Terapia Intensiva , Transferência de PacientesRESUMO
First identified as a high-affinity kinase-deficient receptor for class-3 semaphorins and vascular endothelial growth factor (VEGF) families, Neuropilin2 (NRP2) is a transmembrane non-tyrosine-kinase glycoprotein that has a vital function in neuronal patterning. Furthermore, NRP2 expression is often upregulated in cancer tissues and correlated with poor prognosis. In the present study, we report the establishment of a monoclonal antibody specific for NRP2b1b2 domain (NRP2 MAb) through hybridoma method. NRP2 MAb is measured to have a titer of 5.12 × 10(5) against NRP2b1b2 in indirect ELISA. Western blotting, flow cytometry, and immunofluorescence analysis indicate that NRP2 MAb can combine full-length NRP2 in LoVo and SW480 cells. Besides helping further understand NRP2-related pathological mechanisms and cell-signaling pathways, NRP2 MAb may act as a therapeutic agent for cancer in the future.
Assuntos
Anticorpos Monoclonais/química , Anticorpos Monoclonais/imunologia , Neuropilina-2/imunologia , Animais , Células Cultivadas , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Imunofluorescência/métodos , Hibridomas/imunologia , Proteínas de Membrana/química , Proteínas de Membrana/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Estrutura Terciária de Proteína , Transdução de Sinais/imunologia , Regulação para Cima/imunologia , Fator A de Crescimento do Endotélio Vascular/imunologiaRESUMO
Dextran as anti-nutritional factor is usually a result of bacteria activity and has associated serial problems during the process stream in the sugar industry and in medical therapy. A sensitive method is expected to detect dextran quantitatively. Here we generated four monoclonal antibodies (MAbs) against dextran using dextran T40 conjugated with bovine serum albumin (BSA) as immunogen in our lab following hybridoma protocol. Through pairwise, an MAb named D24 was determined to be conjugated with horseradish peroxidase (HRP) and was used in the establishment of a sensitive sandwich enzyme-linked immunosorbent assay (ELISA) method for determination of dextran, in which MAb D9 was chosen as a capture antibody. The detection limit and working scope of the developed sandwich ELISA method were 3.9 ng/mL and 7.8-500 ng/mL with a correlation coefficient of 0.9909. In addition, the cross-reaction assay demonstrated that the method possessed high specificity with no significant cross-reaction with dextran-related substances, and the recovery rate ranged from 96.35 to 102.00%, with coefficient of variation ranging from 1.58 to 6.94%. These results indicated that we developed a detection system of MAb-based sandwich ELISA to measure dextran and this system should be a potential tool to determine dextran levels.
Assuntos
Anticorpos Monoclonais/imunologia , Dextranos/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Animais , Linhagem Celular , Reações Cruzadas/imunologia , Peroxidase do Rábano Silvestre/imunologia , Hibridomas/imunologia , Limite de Detecção , Camundongos , Sensibilidade e EspecificidadeRESUMO
Naked2 (NKD2) is a member of the Naked family and negatively regulates canonical Wnt signaling. NKD2 may play a role in embryo development and tumor formation by affecting Wnt signaling. In the present study, we describe the establishment of a monoclonal antibody against NKD2 (anti-NKD2 MAb) through the hybridoma method. The purified anti-NKD2 MAb measured a titer of 2.56 × 10(5) against NKD2 by indirect ELISA. Western blot analysis, immunoprecipitation, and confocal microscope showed that the anti-NKD2 MAb can specifically combine NKD2 protein in SW480 and LOVO cells. Competitive inhibition assays of Western blot and indirect ELISA showed that the anti-NKD2 MAb can be blocked with NKD2(1-217) protein. The anti-NKD2 MAb would be helpful for further studies on the structure activity relationship, protein detecting, and cell-signaling pathway of NKD2.
Assuntos
Anticorpos Monoclonais/imunologia , Proteínas de Transporte/imunologia , Neoplasias Colorretais/imunologia , Hibridomas/imunologia , Proteínas Adaptadoras de Transdução de Sinal , Animais , Anticorpos Monoclonais/biossíntese , Anticorpos Monoclonais/isolamento & purificação , Formação de Anticorpos , Especificidade de Anticorpos , Western Blotting , Proteínas de Ligação ao Cálcio , Proteínas de Transporte/isolamento & purificação , Proteínas de Transporte/metabolismo , Neoplasias Colorretais/metabolismo , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Humanos , Imunização , Imunoprecipitação , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Confocal , Proteínas Recombinantes , Células Tumorais CultivadasRESUMO
In the title compound, [Ag(C(7)H(5)O(2))(C(5)H(6)N(2))(2)], the Ag(I) atom is tricoordinated by two independent pyridine N atoms and one benzoate O atom in a nearly planar geometry. An intramolecular N-H.O hydrogen bond forms an S(8) graph ring. The packing is built from molecular layers stabilized by two types of N-H.O hydrogen bond. Intermolecular Ag.N and intramolecular Ag.O contacts were also observed, together with three weak intermolecular C-H.pi interactions.
RESUMO
In the ternary title compound, catena-poly[[silver(I)-mu-ethylenediamine-kappa(2)N:N'] 3-nitrobenzoate monohydrate], [[Ag(C(2)H(8)N(2))](C(7)H(4)NO(4)) x H(2)O](n), the Ag atom is bicoordinated in a linear configuration by two different N atoms from two symmetry-related ethylenediamine ligands, thus giving linear polymeric chains with an [-Ag-N-C-C-N-](n) backbone running parallel to the a axis. In the crystal packing, these linear chains are interconnected by N-H...O and O-H...O hydrogen bonds to form layers parallel to the ab plane.