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This article presents a case study of diffuse large B-cell lymphoma in an elderly patient whose initial clinical manifestation was rapidly developing abdominal distension. The article delves into the patient's diagnostic and treatment journey, highlights treatment insights, and reviews relevant literature. The aim is to enhance the clinical diagnosis accuracy for elderly lymphoma patients presenting with a singular atypical symptom, ultimately optimizing treatment plans and enriching clinicians' knowledge of the disease.
RESUMEN
In the field of genetic code expansion (GCE), improvements in the efficiency of noncanonical amino acid (ncAA) incorporation have received continuous attention. By analyzing the reported gene sequences of giant virus species, we noticed some sequence differences at the tRNA binding interface. On the basis of the structural and activity differences between Methanococcus jannaschii Tyrosyl-tRNA Synthetase (MjTyrRS) and mimivirus Tyrosyl-tRNA Synthetase (MVTyrRS), we found that the size of the anticodon-recognized loop of MjTyrRS influences its suppression activity regarding triplet and specific quadruplet codons. Therefore, three MjTyrRS mutants with loop minimization were designed. The suppression of wild-type MjTyrRS loop-minimized mutants increased by 1.8-4.3-fold, and the MjTyrRS variants enhanced the activity of the incorporation of ncAAs by 15-150% through loop minimization. In addition, for specific quadruplet codons, the loop minimization of MjTyrRS also improves the suppression efficiency. These results suggest that loop minimization of MjTyrRS may provide a general strategy for the efficient synthesis of ncAAs-containing proteins.
Asunto(s)
Aminoácidos , Tirosina-ARNt Ligasa , Aminoácidos/metabolismo , Tirosina-ARNt Ligasa/genética , Tirosina-ARNt Ligasa/metabolismo , Anticodón/genética , Methanocaldococcus/genética , Methanocaldococcus/metabolismo , CodónRESUMEN
The activity and function of proteins can be improved by incorporation of non-canonical amino acids (ncAAs). To avoid the tedious synthesis of a large number of chiral phenylalanine derivatives, we synthesized the corresponding phenylpyruvic acid precursors. Escherichia coli strain DH10B and strain C321.ΔA.expΔPBAD were selected as hosts for phenylpyruvic acid bioconversion and genetic code expansion using the MmPylRS/pyltRNACUA system. The concentrations of keto acids, PLP and amino donors were optimized in the process. Eight keto acids that can be biotransformed and their coupled genetic code expansions were identified. Finally, the genetic encoded ncAAs were tested for incorporation into fluorescent proteins with keto acids.
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Código Genético , Cetoácidos/metabolismo , Fenilalanina/genética , Escherichia coli/metabolismo , Fluorescencia , Proteínas Fluorescentes Verdes/metabolismo , Cetoácidos/química , Fenilalanina/química , Fosfato de Piridoxal/metabolismo , Especificidad por SustratoRESUMEN
Single B cell antibody technology is a method for isolating antigen-specific B cells from human peripheral blood and obtaining antibody genes in developing antibody drugs. However, owing to immune tolerance to autoantigen, human autoantigen-specific B cells are difficult to acquire by conventional single B cell technology. In this study, we constructed a nitrated T-cell epitope named NitraTh by incorporating p-nitrophenylalanine into a universal T helper epitope. NitraTh had enhanced ability to activate CD4+ T cells and can be recognized by CD4+ T cells with different HLA class II haplotypes. This NitraTh can also break immune tolerance to autoantigens, such as human epidermal growth factor receptor 2 (HER2) and cannabinoid receptor 1, and induce strong specific IgM+ B cell responses in vitro. HER2-NitraTh vaccine can also stimulate the generation of HER2-specific IgG+ B cells in human immune system mice, which was established by cotransplanting lymphocytes and autologous dendritic cells in immunodeficient mice. We obtained 30 fully human IgG antibody genes by sorting single B cells from the human immune system mice immunized with HER2-NitraTh vaccine. The analysis of antibody genes showed that sorted B cells underwent the extensive somatic mutation of the antibody genes. We randomly selected eight genes for cloning, six of which expressed antibodies that can bind to HER2. Hence, we provided a convenient and effective method in acquiring fully human antibody genes against self-proteins, which can be used in developing therapeutic antibody drugs.
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Linfocitos B/inmunología , Linfocitos T CD4-Positivos/inmunología , Diferenciación Celular/inmunología , Epítopos de Linfocito T/inmunología , Fenilalanina/análogos & derivados , Linfocitos T Colaboradores-Inductores/inmunología , Animales , Anticuerpos/uso terapéutico , Autoantígenos/inmunología , Vacunas contra el Cáncer/inmunología , Línea Celular Tumoral , Células Dendríticas/inmunología , Epítopos de Linfocito T/química , Femenino , Antígenos de Histocompatibilidad Clase II , Humanos , Tolerancia Inmunológica , Inmunoglobulina G/genética , Inmunoglobulina M , Ratones , Ratones Endogámicos NOD , Fenilalanina/química , Fenilalanina/inmunología , Receptor ErbB-2/inmunologíaRESUMEN
PEG modification is a common clinical strategy for prolonging the half-life of therapeutic proteins or polypeptides. In a previous work, we have successfully synthesized PEG-modified Exendin-4 (PE) by conjugating a 20 kDa PEG to the C-terminal of Exendin-4. Then, we introduced an integrative characterization for PE to evaluate its hypoglycemic activity and pharmacokinetic properties. The normoglycemic efficacies and therapeutic activity of PE were investigated in db/db mice. The hypoglycemic time after single administration of PE on db/db mice was prolonged from 8.4 h to 54.9 h. In multiple treatment with PE, the fasting blood glucose in various PE dosages (50, 150, and 250 nmol/kg) were remarkably reduced, and the glycosylated hemoglobin level was decreased to 2.0%. When the in vivo single- and multiple-dose pharmacokinetics of PE were examined in Sprague-Dawley rats, the half-life was prolonged to 31.7 h, and no accumulation effect was observed. Overall, this study provided a novel promising therapeutic approach to improving glucose-controlling ability and extending half-life without accumulation in vivo for long-acting treatment of type-2 diabetes.
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Preparaciones de Acción Retardada/administración & dosificación , Preparaciones de Acción Retardada/farmacología , Exenatida/administración & dosificación , Exenatida/farmacología , Hipoglucemiantes , Animales , Glucemia/metabolismo , Células Cultivadas , Preparaciones de Acción Retardada/síntesis química , Preparaciones de Acción Retardada/farmacocinética , Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Relación Dosis-Respuesta a Droga , Exenatida/síntesis química , Exenatida/farmacocinética , Resistencia a la Insulina , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos , Ratas Sprague-Dawley , Factores de TiempoRESUMEN
Human epidermal growth factor receptor 2 (HER2) is an attractive target for cancer vaccine. However, autoimmune tolerance prevents vaccines based on HER2 protein from inducing long-lasting, highly effective anti-tumor immunity. In this study, we proved that the introduction of p-nitrophenylalanine in the universal T cell epitope (named NitraTh) enhances humoral immunity induced by B cell epitope and cellular immunity induced by CTL epitope. Moreover, this NitraTh epitope can work in both mouse and human immune system. When fused with extracellular domain 23-83 of HER2, NitraTh epitope help to break the self-tolerance of HER2 and induced strong HER2 specific humoral immunity and cellular immunity. Vaccination with HER2-NitraTh can significantly inhibit the growth of HER2+B16F10 tumor cells. These findings have important implications for developing therapeutic cancer vaccines.
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Neoplasias de la Mama/terapia , Vacunas contra el Cáncer/inmunología , Epítopos de Linfocito T/inmunología , Inmunogenicidad Vacunal , Receptor ErbB-2/inmunología , Animales , Neoplasias de la Mama/inmunología , Neoplasias de la Mama/patología , Vacunas contra el Cáncer/administración & dosificación , Vacunas contra el Cáncer/genética , Línea Celular Tumoral , Epítopos de Linfocito T/química , Epítopos de Linfocito T/genética , Femenino , Humanos , Tolerancia Inmunológica/efectos de los fármacos , Inmunidad Celular/efectos de los fármacos , Inmunidad Celular/inmunología , Inmunidad Humoral/efectos de los fármacos , Inmunidad Humoral/inmunología , Melanoma Experimental/inmunología , Melanoma Experimental/patología , Melanoma Experimental/terapia , Ratones , Nitratos/química , Dominios Proteicos/genética , Dominios Proteicos/inmunología , Receptor ErbB-2/administración & dosificación , Receptor ErbB-2/genética , Proteínas Recombinantes de Fusión/administración & dosificación , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , Linfocitos T Colaboradores-Inductores/inmunología , Resultado del Tratamiento , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunologíaRESUMEN
Human epidermal growth factor receptor 2 (HER2) is a powerful target for cancer immune therapy. The development of anti-HER2 monoclonal antibodies targeting different domains of HER2 is quite effective. However, the selection and production of multivalent antibodies are complicated. In this study, a mimivirus-based designed ankyrin repeat protein (DARPin) targeting HER2 was selected from an artificial library by bacteria surface display. The selection was performed on HER2-positive B16BL6/E2 melanoma cells and HER2-nagative cells. DARPin selected from the library could be expressed in soluble form with a yield of 70 mg/L. After purified by two continuous and easy steps, the purity of DARPin was 90% as established by SDS-PAGE and RP-HPLC. Selected DARPin showed significant HER2-targeting ability with an affinity of 1.05 ± 0.47 µM. MTT assay demonstrated that at the concentration of 640 nM, the selected DARPin dimer could inhibit the SK-BR-3 growth at a rate of 36.63 and 46.34% in 48 and 72 hr incubation separately, which was similar to trastuzumab (43.12 and 49.14% separately). These findings suggested that it was an effective method to select antibody mimetic DARPin by bacteria surface display combined with live cells sorting and provided a drug candidate for cancer therapy.
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Repetición de Anquirina , Melanoma/tratamiento farmacológico , Mimiviridae/metabolismo , Receptor ErbB-2/metabolismo , Proteínas Virales/metabolismo , Proteínas Virales/farmacología , Secuencia de Aminoácidos , Antineoplásicos/química , Antineoplásicos/metabolismo , Antineoplásicos/farmacología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Escherichia coli/metabolismo , Biblioteca de Genes , Humanos , Melanoma/metabolismo , Mimiviridae/química , Modelos Moleculares , Unión Proteica , Proteínas Virales/químicaRESUMEN
Polymer-based paclitaxel (PTX) conjugates have demonstrated application potentials to improve the water solubility and enhance the efficiency of drug delivery. In this study, a novel HA-based drug conjugate, HA-6-PTX, was designed and successfully synthesized by chemically grafting PTX to the C-6 position of N-acetyl-d-glucosamine (GlcNAc) of hyaluronic acid (HA) using hexanediamine as the linker. Leaving the carboxylate of HA chain unaffected, the conjugate with drug loading as high as 21.8% showed an excellent water solubility of 168mg/mL and exhibited increased drug release in the presence of hyaluronidase. Compared to free PTX, HA-6-PTX demonstrated increased cytotoxicity and enhanced apoptosis-inducing effect against HepG2 and A549 cells due to the increased cellular uptake of drug via HA-receptor mediated endocytosis. It was concluded that the HA-6-PTX conjugate could be potentially utilized for further exploration as targeted drug delivery to enhance antitumor efficacy.
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Acetamidas/química , Diaminas/química , Sistemas de Liberación de Medicamentos , Hexanos/química , Ácido Hialurónico/química , Neoplasias/tratamiento farmacológico , Paclitaxel/química , Paclitaxel/uso terapéutico , Acetamidas/síntesis química , Apoptosis/efectos de los fármacos , Espectroscopía de Resonancia Magnética con Carbono-13 , Muerte Celular/efectos de los fármacos , Diaminas/síntesis química , Liberación de Fármacos , Endocitosis/efectos de los fármacos , Citometría de Flujo , Células Hep G2 , Hexanos/síntesis química , Humanos , Ácido Hialurónico/síntesis química , Neoplasias/patología , Paclitaxel/farmacología , Espectroscopía de Protones por Resonancia MagnéticaRESUMEN
Exendin-4 is a strong therapeutic candidate for the treatment of metabolic syndrome. Related receptor agonist drugs have been on the market since 2005. However, technical limitations and the pain caused by subcutaneous injection have severely limited patient compliance. The goal of the study is to investigate a biologically active exendin-4 analog could be administered orally. Using intraperitoneal glucose tolerance tests, we discovered that exendin4-cysteine administered by oral gavage had a distinct hypoglycemic effect in C57BL/6J mice. Using Rosetta Design and Amber, we designed and screened a series of exendin4-cysteine analogs to identify those that retained biological activity while resisting trypsin digestion. Trypsin Cleavage Site Mutated Exendin4-cysteine 1 (TSME-1), an analog whose bioactivity was similar to exendin-4 and was almost completely resistant to trypsin, was screened out. In addition, TSME-1 significantly normalized the blood glucose levels and the availability of TSME-1 was significantly higher than that of exendin-4 and exendin4-cysteine. Collectively orally administered TSME-1, a trypsin-resistant exendin-4 analog obtained by the system, is a strong candidate for future treatments of type 2 diabetes.
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Cisteína/genética , Diseño de Fármacos , Receptor del Péptido 1 Similar al Glucagón/agonistas , Mutación , Péptidos/genética , Péptidos/farmacología , Ponzoñas/genética , Ponzoñas/farmacología , Administración Oral , Animales , Disponibilidad Biológica , Glucemia/efectos de los fármacos , AMP Cíclico/metabolismo , Cisteína/química , Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Diabetes Mellitus Tipo 2/metabolismo , Exenatida , Receptor del Péptido 1 Similar al Glucagón/química , Prueba de Tolerancia a la Glucosa , Intestino Delgado/enzimología , Masculino , Ratones , Péptido Hidrolasas/metabolismo , Péptidos/administración & dosificación , Péptidos/química , Unión Proteica , Proteolisis , Transducción de Señal/efectos de los fármacos , Relación Estructura-Actividad , Tripsina/metabolismo , Ponzoñas/administración & dosificación , Ponzoñas/químicaRESUMEN
Fatty acid synthesis (FAS) is an essential metabolism during the whole growth and development process of the bacterial. Several key enzymes which involved in this biosynthetic pathway have been considered as useful targets for the development of new antibacterial agents. Among them, ß-ketoacyl-acyl carrier protein synthase III (FabH) is the most magnetic target, since it is central to the initiation of fatty acid biosynthesis and is highly conserved of both Gram-positive and Gram-negative bacteria. Following the previous researches, Schiff-based derivatives with dioxygenated rings and N-heterocycle were synthesized in succession, and their biological activities as potential FabH inhibitors were evaluated in this paper. Among these 15 compounds, compound 2E exhibited the best antibacterial activities with minimum inhibitory concentration (MIC) values 1.56-3.13 mg/mL against the tested bacterial strains and showed the most powerful Escherichia coli (E. coli) FabH inhibitory activities with IC50 of 2.1 µM. Also the conceivable binding conformation of placing compound 2E into the E. coli FabH active site was affirmed docking simulation.
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3-Oxoacil-(Proteína Transportadora de Acil) Sintasa/antagonistas & inhibidores , Antibacterianos/química , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Compuestos Heterocíclicos/química , Compuestos de Nitrógeno/química , Bases de Schiff/química , Antibacterianos/farmacología , Compuestos Heterocíclicos/farmacología , Pruebas de Sensibilidad Microbiana , Compuestos de Nitrógeno/farmacología , Bases de Schiff/farmacología , Relación Estructura-ActividadRESUMEN
In the present work, a simple and sensitive turn-on fluorescence method for DNA detection was developed. It was explored based on the N-methyl mesoporphyrin IX (NMM)/G-quadruplex DNA system as a reporter and exonuclease III (Exo III)-aided target recycling amplification to ensure sensitivity. Our method showed an ultra-wide detection range from 10 fM to 100 nM with a low linear detection limit of 0.76 nM. It also had excellent selectivity in a selectivity experiment.
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Técnicas Biosensibles/métodos , ADN/análisis , Exodesoxirribonucleasas/metabolismo , Límite de Detección , Técnicas de Amplificación de Ácido Nucleico , ADN/química , ADN/metabolismo , G-Cuádruplex , Mesoporfirinas/química , Espectrometría de FluorescenciaRESUMEN
Sortase mediated ligation is a highly specific platform for conjugation that relies on the specificity of the transpeptidase Sortase A (SrtA) for short peptide sequences (LPXTG and GGG). SrtA retains its specificity while accepting a wide range of potential substrates, but its broad use is limited by the wild-type enzyme's poor kinetics, which require large amounts of SrtA and extended reaction times for efficient conjugation. Prior explorations have aimed to improve the kinetics of SrtA with limited success. Herein we describe the discovery of further improved SrtA variants with increased efficiency for the conjugation reaction, and demonstrate their robustness in labelling proteins and antibodies in a site-specific manner. Our variants require significantly lower amounts of enzyme than WT SrtA and can be used to attach small molecules to the N or C-terminus of the heavy or light chain in antibodies with excellent yields. These improved variants can also be used for highly efficient site-specific PEGylation.
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Aminoaciltransferasas , Anticuerpos/química , Proteínas Bacterianas , Cisteína Endopeptidasas , Mutación Missense , Polietilenglicoles/química , Coloración y Etiquetado/métodos , Sustitución de Aminoácidos , Aminoaciltransferasas/química , Aminoaciltransferasas/genética , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Cisteína Endopeptidasas/química , Cisteína Endopeptidasas/genéticaRESUMEN
OBJECTIVES: To prepare recombinant tPep-(vascular endothelial growth factor) VEGF-B and assess its biological activity. RESULTS: This new VEGF fusion protein was constructed using a targeting peptide and prepared using E.coli. The tPep-VEGF-B was refolded from inclusion bodies and purified using affinity chromatography. Its bioactivity was determined in vitro using proliferation assay and wounding healing assay, and in vivo in zebrafish. By using the optimized downstream process, recombinant tPep-VEGF-B can be obtained with a purity of >90 % and a yield of 80 mg protein/l culture medium. The refolded protein is highly effective in promoting cell migration in vitro and in enhancing angiogenesis in vivo. CONCLUSION: We have constructed a new VEGF fusion protein with potential therapeutic application in treating metabolic diseases.
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Proteínas Recombinantes de Fusión/metabolismo , Proteínas Recombinantes/metabolismo , Animales , Línea Celular , Proliferación Celular/efectos de los fármacos , Humanos , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/farmacología , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacología , Cicatrización de Heridas/efectos de los fármacos , Pez CebraRESUMEN
Fatty acid biosynthesis is essential for bacterial survival. ß-Ketoacyl-acyl carrier protein (ACP) synthase III (FabH), is a particularly attractive antibacterial target, since it is central to the initiation of fatty acid biosynthesis. Three series of 21 cinnamaldehyde acylhydrazone derivatives, A3-9, B3-9, and C3-9, were synthesized and evaluated for FabH-inhibitory activity. Compound B6 showed the most potent biological activity against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus subtilis (minimum inhibitory concentrations (MICs) values: 1.56-3.13 µg/mL) and was comparable with the positive control. Docking simulation by positioning compound B6 in the FabH structure active site was performed to explore the possible binding model.
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3-Oxoacil-(Proteína Transportadora de Acil) Sintasa/antagonistas & inhibidores , Acroleína/análogos & derivados , Antibacterianos/química , Bacterias/efectos de los fármacos , Bacterias/enzimología , Dioxanos/química , Hidrazonas/química , 3-Oxoacil-(Proteína Transportadora de Acil) Sintasa/metabolismo , Acroleína/síntesis química , Acroleína/química , Acroleína/farmacología , Antibacterianos/síntesis química , Antibacterianos/farmacología , Bacillus subtilis/efectos de los fármacos , Bacillus subtilis/enzimología , Infecciones Bacterianas/tratamiento farmacológico , Infecciones Bacterianas/enzimología , Infecciones Bacterianas/microbiología , Dioxanos/síntesis química , Dioxanos/farmacología , Diseño de Fármacos , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Escherichia coli/efectos de los fármacos , Escherichia coli/enzimología , Humanos , Hidrazonas/síntesis química , Hidrazonas/farmacología , Simulación del Acoplamiento Molecular , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/enzimología , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/enzimología , Relación Estructura-ActividadRESUMEN
We report the design and synthesis of a series of new cyclometalated iridium hydrides derived from the C-H bond activation of aromatic nitrones and the biological evaluation of these iridium hydrides as antitumor agents. The nitrone ligands are based on the structure of a popular antioxidant, α-phenyl-N-tert-butylnitrone (PBN). Compared to cisplatin, the iridium hydrides exhibit excellent antitumor activity on HepG2 cells. The metal-coordinated compound with the most potent anticancer activity, 2f, was selected for further analysis because of its ability to induce apoptosis and interact with DNA. During in vitro studies and in vivo efficacy analysis with tumor xenograft models in Institute of Cancer Research (ICR) mice, complex 2f exhibited antitumor activity that was markedly superior to that of cisplatin. Our results suggest, for the first time, that metal hydrides could be a new type of metal-based antitumor agent.