RESUMEN
CD19 CAR-T (chimeric antigen receptor-T) cell immunotherapy achieves a remission rate of approximately 70% in recurrent and refractory lymphoma treatment. However, the loss or reduction of CD19 antigen on the surface of lymphoma cells results in the escape of tumor cells from the immune killing of CD19 CAR-T cells (CAR19-T). Therefore, novel therapeutic strategies are urgently required. In this study, an anti-CD79b/CD3 bispecific antibody (BV28-OKT3) was constructed and combined with CAR19-T cells for B-cell lymphoma treatment. When the CD19 antigen was lost or reduced, BV28-OKT3 redirected CAR19-T cells to CD79b+ CD19- lymphoma cells; therefore, BV28-OKT3 overcomes the escape of CD79b+ CD19- lymphoma cells by the killing action of CAR19-T cells in vitro and in vivo. Furthermore, BV28-OKT3 triggered the antitumor function of CAR- T cells in the infusion product and boosted the antitumor immune response of bystander T cells, markedly improving the cytotoxicity of CAR19-T cells to lymphoma cells in vitro and in vivo. In addition, BV28-OKT3 elicited the cytotoxicity of donor-derived T cells toward lymphoma cells in vitro, which depended on the presence of tumor cells. Therefore, our findings provide a new clinical treatment strategy for recurrent and refractory B-cell lymphoma by combining CD79b/CD3 BsAb with CAR19-T cells.
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Anticuerpos Biespecíficos , Linfoma de Células B , Linfoma , Humanos , Linfocitos T , Antígenos CD19 , Muromonab-CD3 , Linfoma/tratamiento farmacológico , Inmunoterapia Adoptiva/métodosRESUMEN
Recombinant protein drugs, which are typically produced by mammalian host cells, have been approved for the treatment of a range of diseases. Accordingly, systems for selecting recombinant cell lines with efficient protein expression and for testing the content of recombinant proteins in vivo are crucial to the large-scale production and application of protein-based therapeutic drugs. In this study, we designed three aptamer beacons to detect His-tag, a common label of recombinant proteins. We found that all three beacons could specifically and quantitatively measure the His-tagged recombinant proteins with a short reaction time. Among these three beacons, the 6H5-MU beacon had the highest sensitivity for His polypeptides with a detection limit of 250 ng/mL and the shortest detection time within 1 min. Furthermore, we established a rapid and highly effective recombinant cell line construction system, which could obtain monoclonal cell lines with high yields of target proteins within 21 days, by combining 6H5-MU with pSB, a novel plasmid composed of a Sleeping Beauty transposase and a transposon. Finally, 6H5-MU also discriminately tested the serum concentration of His-tagged recombinant proteins in vivo, with consistent results compared to enzyme-linked immunosorbent assay (ELISA). We thus established a rapid and high-throughput method for generating recombinant cell lines and in vivo monitoring of recombinant protein levels, thereby providing a new platform for the development and preparation of recombinant protein drugs. KEY POINTS: ⢠The 6H5-MU aptamer beacon rapidly and accurately binds to His-tagged recombinant proteins. ⢠A system for rapid and high-throughput generation of recombinant cell lines is established using 6H5-MU and pSB. ⢠6H5-MU allows in vivo monitoring of recombinant protein levels.
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Mamíferos , Oligonucleótidos , Animales , Proteínas Recombinantes/genética , Línea CelularRESUMEN
With the in-depth study of circRNA, more and more biological studies have shown that circRNAs play an important role in mammals, such as cell proliferation, apoptosis, invasion, development and disease state. However, the regulatory mechanism of circRNA in lower vertebrates remains unclear. Here, we found a new circular RNA and named it circRara. We carried out the experimental study on its antiviral and antibacterial response, cell proliferation and activity. The results showed that circRara had a positive regulatory effect on the antiviral and antibacterial response, cell proliferation and activity in miiuy croaker. First, we found that the expression of circRara could be up-regulated under the stimulation of LPS and poly (I: C), but not the expression of linear Rara. In addition, the increase of circRara can increase the production of inflammatory factors and antiviral genes, which was confirmed by double luciferase reporter gene experiment and qPCR. These results will help to further understand the immunomodulatory mechanism of circRNA in teleost fish.
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Perciformes , Vibriosis , Secuencia de Aminoácidos , Animales , Antibacterianos , Antivirales , Proteínas de Peces/genética , Proteínas de Peces/metabolismo , Inmunidad Innata/genética , Lipopolisacáridos/farmacología , Mamíferos/genética , Mamíferos/metabolismo , Filogenia , ARN Circular/genética , Alineación de SecuenciaRESUMEN
BACKGROUND: Diabetes is associated with endothelial cell dysfunction. E-selectin is an endothelial cell adhesion molecule, which is bound for endothelial cell activation. E-selectin gene+A561C polymorphism is associated with many different disorders: essential hypertension, stroke, angina pectoris, coronary heart disease, etc. But the association with type 2 diabetes remains unclear. Therefore, we aimed to investigate the role of E-selectin gene+A561C polymorphism and soluble E-selectin in type 2 diabetes in a Chinese population. METHODS: This study involved 317 patients with type 2 diabetes and 285 normal healthy controls. Genotyping of E-selectin gene+A561C polymorphism was examined by polymerase chain reaction-restricted fragments length polymorphism (PCR-RFLP). Soluble E-selectin was examined by enzyme linked immunosorbent assay (ELISA). Biochemical markers were measured by Roche 7600 Automated Biochemical Analyzer. RESULTS: We found that C allele frequency in E-selectin A561 C polymorphism of Chinese T2DM group was higher than control group. The level of soluble E-selectin in T2DM group was higher than control group. TC, TG, LDL-C, ApoB, and sE-selectin (soluble E-selectin) in AC and CC genotypes were higher than AA genotype. CONCLUSIONS: Our findings showed that E-selectin +A561C polymorphism was correlated in the Chinese population with type 2 diabetes. C allele and soluble E-selectin may be predisposing factors of Chinese population with type 2 diabetes.
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Diabetes Mellitus Tipo 2 , Selectina E , China , Diabetes Mellitus Tipo 2/genética , Selectina E/genética , Selectina E/metabolismo , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Polimorfismo GenéticoRESUMEN
During human erythroid maturation, Hsp70 translocates into the nucleus and protects GATA-1 from caspase-3 cleavage. Failure of Hsp70 to localize to the nucleus was found in Myelodysplastic syndrome (MDS) erythroblasts and can induce dyserythropoiesis, with arrest of maturation and death of erythroblasts. However, the mechanism of the nuclear trafficking of Hsp70 in erythroblasts remains unknown. Here, we found the hematopoietic transcriptional regulator, EDAG, to be a novel binding partner of Hsp70 that forms a protein complex with Hsp70 and GATA-1 during human normal erythroid differentiation. EDAG overexpression blocked the cytoplasmic translocation of Hsp70 induced by EPO deprivation, inhibited GATA-1 degradation, thereby promoting erythroid maturation in an Hsp70-dependent manner. Furthermore, in myelodysplastic syndrome (MDS) patients with dyserythropoiesis, EDAG is dramatically down-regulated, and forced expression of EDAG has been found to restore the localization of Hsp70 in the nucleus and elevate the protein level of GATA-1 to a significant extent. In addition, EDAG rescued the dyserythropoiesis of MDS patients by increasing erythroid differentiation and decreasing cell apoptosis. This study demonstrates the molecular mechanism of Hsp70 nuclear sustaining during erythroid maturation and establishes that EDAG might be a suitable therapeutic target for dyserythropoiesis in MDS patients.
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Núcleo Celular/metabolismo , Eritroblastos/metabolismo , Eritropoyesis/fisiología , Proteínas HSP70 de Choque Térmico/metabolismo , Síndromes Mielodisplásicos/metabolismo , Proteínas Nucleares/metabolismo , Apoptosis/fisiología , Caspasa 3/metabolismo , Diferenciación Celular/fisiología , Células Cultivadas , Citoplasma/metabolismo , Regulación de la Expresión Génica/fisiología , Enfermedades Hematológicas/metabolismo , HumanosRESUMEN
BACKGROUND: The performance of 17 routine chemical detection methods was evaluated by the Sigma (σ) index, and separate quality control standards were established according to the sigma values of different detection methods. METHODS: The internal quality control (IQC) and external quality assessment (EQA) data of 17 assays in the biochemical laboratory of our hospital were collected from January to June 2019. Referring to the total allowed error (TEa) standards established in the Health Industry Standards of the People's Republic of China (WS/T 403-2012), the sigma metric of each assay was calculated, the performance level for inspection was evaluated, the quality goal index (QGI) was calculated for items with analysis performance < 5 sigma, and the main causes of poor performance were determined to guide quality improvement. RESULTS: For level 1 internal quality control (IQC), five assays (AMY, Crea, UA, TP, and Na) showed a performance of ≥ 6 sigma levels. Five assays (GGT, LDH, ALP, K, and Ca) had a performance lower than 3 sigma. For level 2 IQC, nine assays (ALT, AST, CK, AMY, Crea, UA, TP, Na, and Mg) achieved 6 sigma, and four assays (GGT, LDH, ALP, and K) achieved less than 3 sigma. Among the 12 assays with a sigma value < 5, the precision of 1 assay should be improved first, the accuracy of 6 assays should be improved next, and both the precision and the accuracy of 5 assays should be improved. CONCLUSIONS: The sigma metric is the best tool for evaluating the performance of different test methods. Assays with high sigma values can be evaluated with single-rule quality control, while assays with low values should be evaluated with strict quality control rules.
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Servicios de Laboratorio Clínico , Laboratorios , Gestión de la Calidad Total , China , Humanos , Control de CalidadRESUMEN
BACKGROUND Acute lymphocytic leukemia (ALL) is a common blood cancer which induces high mortality in children. Bromodomains and extra-terminal (BET) protein inhibitors, such as JQ1 and ARV-825, are promising cancer therapeutic agents that can be used by targeting c-Myc. A recent work reported that JQ1 effectively attenuates ALL in vitro by suppressing cell proliferation and accelerating apoptosis. The purpose of this research was to probe into the potential mechanism of how JQ1 inhibits ALL cell proliferation in vitro. MATERIAL AND METHODS Cell viability of ALL cells were measured by CTG after treatment by JQ1. Cell cycle analysis was done by EdU and PI staining. Cell apoptosis was assessed by Annexin V/PI staining. Glycolysis was detected using Seahorse and LC-MS kits. The expression of glycolytic rate-limiting enzymes was assessed by RNA-seq, qRT-PCR, and Western blot. RESULTS JQ1 suppressed cell proliferation by arresting the cell cycle and inducing the apoptosis of acute lymphocytic leukemia cells. JQ1 inhibited cell proliferation of B-ALL cells by restraining glycolysis. Conversely, the cell cycle block of B-ALL cells induced by JQ1 was partially abolished after pretreatment with 2-Deoxy-D-glucose (2-DG), an inhibitor of glycolysis. Furthermore, JQ1 restrained the glycolysis of B-ALL cell lines by remarkably downregulating the rate-limiting enzymes of glycolysis, such as hexokinase 2, phosphofructokinase, and lactate dehydrogenase A. Moreover, the cell cycle arrest was reversed in B-ALL cells with overexpressed c-Myc treated by JQ1, which is involved in the enhancement of glycolysis. CONCLUSIONS The BET inhibitor JQ1 suppresses the proliferation of ALL by inhibiting c-Myc-mediated glycolysis, thus providing a new strategy for the treatment of ALL.
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Azepinas/farmacología , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamiento farmacológico , Proteínas/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-myc/antagonistas & inhibidores , Triazoles/farmacología , Apoptosis/efectos de los fármacos , Ciclo Celular/efectos de los fármacos , Puntos de Control del Ciclo Celular/efectos de los fármacos , Proteínas de Ciclo Celular/metabolismo , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Glucólisis/efectos de los fármacos , Células HEK293 , Humanos , Proteínas Nucleares/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/patología , Proteínas/metabolismo , Proteínas Proto-Oncogénicas c-myc/metabolismo , Factores de Transcripción/metabolismoRESUMEN
Aplastic anemia (AA) is a serious blood system disease that threatens human health. At present, the main cause of this disease is believed to be immune hyperfunction. However, the specific metabolic mode involved in the occurrence of lymphocytes in AA is still unknown. In addition, whether rapamycin, a specific blocker of the mTOR signaling pathway, plays a therapeutic role by inhibiting lymphocyte metabolism remains unclear. We induced an AA mouse model through the classical immune-mediated pathway and simultaneously administered rapamycin intervention therapy. First, the AA-associated phenotypic changes and the efficacy of rapamycin in the treatment of AA were discussed. Second, the proliferation and metabolic pathway of bone marrow (BM) lymphocytes in AA and the effect of rapamycin on this process were determined. Finally, the expression levels of mTOR pathway-related proteins were analyzed. By inhibiting the mTOR signaling pathway, rapamycin could ameliorate the phenotype of the immune-mediated AA model and inhibit the proliferation of T cells by preventing cell cycle transition from G0 to G1 phase. Moreover, we found that mitochondrial oxidative phosphorylation is involved in the metabolic reprogramming of T cells in AA and that rapamycin can inhibit this process. We confirmed that mitochondrial oxidative phosphorylation is involved in the metabolic reprogramming of T cells in AA and further extended the mechanism of rapamycin in treating AA by inhibiting the mTOR signaling pathway. This viewpoint may provide a new therapeutic idea for clinical applications.
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Anemia Aplásica/tratamiento farmacológico , Inmunosupresores/farmacología , Sirolimus/farmacología , Linfocitos T/efectos de los fármacos , Anemia Aplásica/inmunología , Animales , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Masculino , Ratones , Ratones Congénicos , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Mitocondrias/patología , Linfocitos T/inmunología , Linfocitos T/patologíaRESUMEN
Leucine-rich repeat containing G-protein-coupled receptor 6 (LGR6) is a member of the rhodopsin-like 7-transmembrane domain receptor superfamily and has high homology to LGR4 and LGR5. LGR6 is highly expressed in osteoblastic progenitors, and LGR6-deficient mice show nail and bone regeneration defect. However, the effect of LGR6 on the osteogenic differentiation of osteoblastic progenitors and its underlying mechanisms are largely unknown. In this study, we overexpressed and knockdown LGR6 with lentivirus in the preosteoblastic cell MC3T3-E1 to observe the effect of LGR6 on osteogenic differentiation and explore its possible molecular mechanism. LGR6 overexpression promoted osteogenic differentiation and mineralization by stabilizing ß-catenin to potentiate the Wnt/ß-catenin signaling pathway in MC3T3-E1 cells. Conversely, LGR6 knockdown inhibited osteogenic differentiation and mineralization by enhancing ß-catenin degradation to inactivate the Wnt/ß-catenin signaling pathway. These results reveal that LGR6 is highly expressed in osteoblastic progenitors, and promotes osteogenesis by enhancing ß-catenin stability to strengthen the Wnt signaling pathway. This study provides an important reference into the exact mechanisms of osteogenic differentiation.
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Osteogénesis , Receptores Acoplados a Proteínas G/metabolismo , Vía de Señalización Wnt , Animales , Calcificación Fisiológica , Diferenciación Celular , Línea Celular , Técnicas de Silenciamiento del Gen , Ratones , Estabilidad Proteica , Proteolisis , beta Catenina/metabolismoRESUMEN
The auditory system has the ability to adjust its structure and function as the environment changes, which is called auditory plasticity. In the auditory system, inferior colliculus (IC) is an important relay station, which accepts the ascending inputs from dorsal nuclei of lateral lemniscus (DNLL). The present study was aimed to investigate the role of the DNLL in the formation of the plasticity of IC neurons. Here, we used extracellular single unit recording and electrical stimulation to investigate the plasticity of IC neurons in Kunming mice. The results showed that after the cessation of 30-minute electrical stimulation on contralateral DNLL, 95% of the inhibited IC neurons and 86% of the facilitated IC neurons showed plastic changes. Moreover, 1 h after the contralateral DNLL stimulation was stopped, the plastic changes in 74% of the inhibited IC neurons vanished, but still were maintained in 26% of the inhibited IC neurons. These results suggest that the contralateral DNLL ascending input can induce plastic changes of IC neurons, and this kind of effect can be maintained for a certain period of time, which is beneficial to enhance the sound intensity sensitivity of IC neurons.
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Vías Auditivas , Colículos Inferiores/fisiología , Plasticidad Neuronal , Neuronas/fisiología , Animales , Estimulación Eléctrica , RatonesRESUMEN
Cholinergic receptor activation and intracellular current injection lead to the persistent activity (PA), which may be involved in inducing neural plasticity. Our previous study showed that PA is closely related to the activation of muscarinic acetylcholine receptors (mAChRs) in pyramidal neurons of mouse primary auditory cortex (AI). However, the subtypes of mAChRs involved in PA remain unclear. Thus, using whole-cell patch-clamp recording and pharmacological methods, we investigated the role of different mAChR subtypes in inducing PA in AI layer V pyramidal neurons of young mice. The results showed that activation of mAChRs with intracellular depolarizing current induced PA in layer V pyramidal neurons. Blockade of M1, M2 or M3 subtypes prevented the PA, whereas M4 receptor antagonists did not affect the production of PA. The results suggest that the PA may be induced through a mechanism involving M1, M2 and M3 muscarinic receptors, but not M4 subtype.
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Corteza Auditiva/fisiología , Células Piramidales/fisiología , Receptores Muscarínicos/fisiología , Acetilcolina , Animales , Ratones , Técnicas de Placa-Clamp , Receptores Muscarínicos/clasificaciónRESUMEN
Erythroid differentiation-associated gene (EDAG) has been considered to be a transcriptional regulator that controls hematopoietic cell differentiation, proliferation, and apoptosis. The role of EDAG in erythroid differentiation of primary erythroid progenitor cells and in vivo remains unknown. In this study, we found that EDAG is highly expressed in CMPs and MEPs and upregulated during the erythroid differentiation of CD34(+) cells following erythropoietin (EPO) treatment. Overexpression of EDAG induced erythroid differentiation of CD34(+) cells in vitro and in vivo using immunodeficient mice. Conversely, EDAG knockdown reduced erythroid differentiation in EPO-treated CD34(+) cells. Detailed mechanistic analysis suggested that EDAG forms complex with GATA1 and p300 and increases GATA1 acetylation and transcriptional activity by facilitating the interaction between GATA1 and p300. EDAG deletion mutants lacking the binding domain with GATA1 or p300 failed to enhance erythroid differentiation, suggesting that EDAG regulates erythroid differentiation partly through forming EDAG/GATA1/p300 complex. In the presence of the specific inhibitor of p300 acetyltransferase activity, C646, EDAG was unable to accelerate erythroid differentiation, indicating an involvement of p300 acetyltransferase activity in EDAG-induced erythroid differentiation. ChIP-PCR experiments confirmed that GATA1 and EDAG co-occupy GATA1-targeted genes in primary erythroid cells and in vivo. ChIP-seq was further performed to examine the global occupancy of EDAG during erythroid differentiation and a total of 7,133 enrichment peaks corresponding to 3,847 genes were identified. Merging EDAG ChIP-Seq and GATA1 ChIP-Seq datasets revealed that 782 genes overlapped. Microarray analysis suggested that EDAG knockdown selectively inhibits GATA1-activated target genes. These data provide novel insights into EDAG in regulation of erythroid differentiation.
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Diferenciación Celular/fisiología , Proteína p300 Asociada a E1A/metabolismo , Factor de Transcripción GATA1/metabolismo , Hematopoyesis/fisiología , Células Madre Hematopoyéticas/citología , Proteínas Nucleares/metabolismo , Acetilación , Animales , Western Blotting , Separación Celular , Células Eritroides/citología , Células Eritroides/metabolismo , Femenino , Citometría de Flujo , Células Madre Hematopoyéticas/metabolismo , Xenoinjertos , Humanos , Ratones , Ratones Endogámicos NOD , Ratones SCID , Análisis de Secuencia por Matrices de Oligonucleótidos , TranscriptomaRESUMEN
Antibody (Ab)-based drugs have been widely used in targeted therapies and immunotherapies, leading to significant improvements in tumor therapy. However, the failure of Ab therapy due to the loss of target antigens or Ab modifications that affect its function limits its application. In this study, we expanded the application of antibodies (Abs) by constructing a fusion protein as a versatile tool for Ab-based target cell detection, delivery, and therapy. We first constructed a SpaC Catcher (SpaCC for short) fusion protein that included the C domains of Staphylococcal protein A (SpaC) and the SpyCatcher. SpaCC conjugated with SpyTag-X (S-X) to form the SpaCC-S-X complex, which binds non-covalently to an Ab to form the Ab-SpaCC-S-X protein complex. The "X" can be a variety of small molecules such as fluoresceins, cell-penetrating peptide TAT, Monomethyl auristatin E (MMAE), and DNA. We found that Ab-SpaCC-S-FITC(-TAT) could be used for target cell detection and delivery. Besides, we synthesized the Ab-SpaCC-SN3-MMAE complex by linking Ab with MMAE by SpaCC, which improved the cytotoxicity of small molecule toxins. Moreover, we constructed an Ab-DNA complex by conjugating SpaCC with the aptamer (Ap) and found that Ab-SpaCC-SN3-Ap boosted the tumor-killing function of T-cells by retargeting tumor cells. Thus, we developed a multifunctional tool that could be used for targeted therapies and immunotherapies, providing a cheap and convenient novel drug development strategy.
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Péptidos de Penetración Celular , Neoplasias , Humanos , Neoplasias/genética , Neoplasias/terapia , Inmunoterapia , Anticuerpos , ADN , Línea Celular TumoralRESUMEN
Chimeric antigen receptor (CAR)-positive cell therapy, specifically with anti-CD19 CAR-T (CAR19-T) cells, achieves a high complete response during tumor treatment for hematological malignancies. Large-scale production and application of CAR-T therapy can be achieved by developing efficient and low-cost enrichment methods for CAR-T cells, expansion monitoring in vivo, and overcoming tumor escape. Here, novel CAR-specific binding aptamers (CAR-ap) to traceless sort CAR-positive cells and obtain a high positive rate of CAR19-T cells is identified. Additionally, CAR-ap-enriched CAR19-T cells exhibit similar antitumor capacity as CAR-ab (anti-CAR antibody)-enriched CAR-T cells. Moreover, CAR-ap accurately monitors the expansion of CAR19-T cells in vivo and predicts the prognosis of CAR-T treatment. Essentially, a novel class of stable CAR-ap-based bispecific circular aptamers (CAR-bc-ap) is constructed by linking CAR-ap with a tumor surface antigen (TSA): protein tyrosine kinase 7 (PTK7) binding aptamer Sgc8. These CAR-bc-aps significantly enhance antitumor cytotoxicity with a loss of target antigens by retargeting CAR-T cells to the tumor in vitro and in vivo. Overall, novel CAR-aptamers are screened for traceless enrichment, monitoring of CAR-positive cells, and overcoming tumor cell immune escape. This provides a low-cost and high-throughput approach for CAR-positive cell-based immunotherapy.
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Receptores Quiméricos de Antígenos , Escape del Tumor , Linfocitos T , Inmunoterapia Adoptiva/métodos , InmunoterapiaRESUMEN
BACKGROUND: Randomized controlled trials (RCTs) have reported different results when using zoledronate to treat skeletal-related events (SREs) and bone pain in patients with metastatic bone disease (MBD), and few have looked at the risks and benefits of long-term use of the drug. This meta-analysis aimed to investigate the efficacy and safety of zoledronate to treat MBD in the short and long-term. METHODS: PubMed, EMBASE, and the Cochrane Library were searched to identify RCTs evaluating zoledronate for MBD. Relative risks (RRs) and 95% confidence intervals (CIs) were calculated. RESULTS: Twelve RCTs involving 4,450 patients were included in the meta-analysis. Zoledronate decreased the risk of developing SREs compared with placebo (RR 0.75, 95% CI 0.69 to 0.81, P < 0.001). Zoledronate consistently reduced the brief pain inventory (BPI) below baseline compared with placebo at 3, 12, and 24 months. In addition, the likelihood of experiencing a bone pain event was significantly lower in the zoledronate group than in the placebo group (RR 0.83, 95% CI 0.76 to 0.89, P < 0.001). While the two groups did not differ significantly in the incidence of nausea(RR = 1.07, 95% CI 0.96 to 1.19, P = 0.250), emesis (RR 0.94, 95% CI 0.81 to 1.09, P = 0.420), or adverse renal events (RR 1.41, 95% CI 0.94 to 2.11, P = 0.09), the zoledronate group showed a significantly higher relative risk of pyrexia (RR 1.43, 95% CI 1.20 to 1.70, P < 0.001), fatigue (RR 1.26, 95% CI 1.10 to 1.43, P < 0.001), and anemia (RR 1.33, 95% CI 1.14 to 1.55, P < 0.001). CONCLUSION: Compared to placebo, zoledronate significantly reduced the incidence of bone pain and SREs in patients with MBD for periods as long as 24 months. In addition, zoledronate is generally well tolerated over this long period.
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Conservadores de la Densidad Ósea/uso terapéutico , Neoplasias Óseas/secundario , Difosfonatos/uso terapéutico , Fracturas Espontáneas/prevención & control , Imidazoles/uso terapéutico , Dolor Musculoesquelético/tratamiento farmacológico , Compresión de la Médula Espinal/prevención & control , Neoplasias Óseas/complicaciones , Humanos , Dolor Musculoesquelético/etiología , Dimensión del Dolor , Ensayos Clínicos Controlados Aleatorios como Asunto , Resultado del Tratamiento , Ácido ZoledrónicoRESUMEN
Antibody drugs have been widely used to treat many diseases and are the fastest-growing drug class. IgG1 is the most common type of antibody because of its good serum stability; however, effective methods for the rapid detection of IgG1-type antibodies are lacking. In this study, we designed two aptamer molecules derived from the reported aptamer probe that has been proven to bind to the Fc fragment of the IgG1 antibody. The results showed that Fc-1S could specifically bind to the human IgG1 Fc proteins. In addition, we modified the structure of Fc-1S and constructed three aptamer molecular beacons that could quantitatively detect IgG1-type antibodies within a short time. Furthermore, we unveiled that the Fc-1S37R beacon has the highest sensitivity for IgG1-type antibodies with a detection limit of 48.82813 ng/mL and can accurately detect serum antibody concentrations in vivo with consistent results to ELISA. Therefore, Fc-1S37R is an efficient method for the production monitoring and quality control of IgG1-type antibodies to enable the large-scale production and application of antibody drugs.
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Fragmentos Fc de Inmunoglobulinas , Inmunoglobulina G , Humanos , Inmunoglobulina G/química , Fragmentos Fc de Inmunoglobulinas/químicaRESUMEN
AIM: To describe the optical coherence tomography angiography (OCTA) characteristics of exudative and non-exudative treatment-naïve pachychoroid neovasculopathy (PNV). METHODS: Thirty-five patients with exudative treatment-naïve PNV and 13 with non-exudative treatment-naïve PNV between March 2020 and December 2021 were included. All patients underwent ophthalmologic examination, including fluorescein angiography (FA), indocyanine green angiography (ICGA), spectral-domain OCT, and OCTA. The clinical data of the patients were retrospectively analyzed. RESULTS: The study included 51 eyes from 46 patients, of whom 33 (71.7%) were male. The central macular thickness (CMT) in the exudative PNV group was significantly higher than that in the non-exudative PNV group (383.97±132.16 µm vs 213.13±51.63 µm; P<0.001). The maximum height of flat irregular pigment epithelial detachments (FIPED) was 45.40±11.86 µm in the non-exudative PNV group, significantly lower than the 71.58±20.91 µm (P<0.001) in the exudative PNV group. The area of PNV of the non-exudative PNV group was, significantly larger than that of the exudative PNV group (1.06±0.84 mm2 vs 0.63±0.80 mm2, P=0.016). There was a significant difference in PNV morphology between the two groups (P<0.001). Multivariate logistic regression analysis found that the maximum height of FIPED (OR=1.156, 95%CI: 1.019-1.312; P=0.024) and microvascular branches (OR=69.412, 95%CI: 3.538-1361.844; P=0.005) were independent predictors of PNV activity. CONCLUSION: The OCTA imaging finds that there are significant differences in CMT, maximum height of FIPED, PNV area, and morphology of exudative PNV and non-exudative PNV groups. OCTA can accurately identify the clinical and imaging features of exudative and non-exudative treatment-naïve PNV, and distinguish PNV activity.
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AIM: To evaluate the efficacy of retinal laser photocoagulation and intravitreal injection of anti-vascular endothelial growth factor (anti-VEGF) for hemorrhagic retinal arterial macroaneurysm (RAM). METHODS: This was a retrospective clinical study. Patients with hemorrhagic RAM were divided into 4 groups defined by different treatments: a retinal laser photocoagulation therapy monotherapy group, an anti-VEGF intravitreal injection monotherapy group, a laser and anti-VEGF combination therapy group, and an observation group. Visual acuity (VA), central macular thickness (CMT), and retinal hemorrhage area (RHA) were collected. RESULTS: Forty-seven eyes of 47 patients were enrolled. VA improved and had a significant difference between baseline and final in each treatment group (logMAR; laser group: 1.90±0.53 vs 1.05±0.63, P<0.001; anti-VEGF group: 1.75±0.63 vs 1.12±0.54, P=0.009; combination group: 1.76±0.38 vs 1.01±0.52, P<0.001); however, VA decreased and had no significant difference in observation group (1.63±0.51 vs 1.76±0.61, P=0.660). CMT decreased and had a significant difference between baseline and final in each group (laser group: 815.16±310.83 vs 252.05±83.90 µm, P<0.001; anti-VEGF group: 725.00±290.79 vs 203.56±69.89 µm, P=0.001; combination group: 595.50±186.51 vs 253.13±55.06 µm, P=0.001; observation group: 758.88±195.65 vs 267.00±120.90 µm, P=0.001). RHA were 28.99±28.15, 25.94±11.58, 19.64±8.97, and 27.45±13.76 mm2 in laser group, anti-VEGF group, combination group and observation group, respectively. RHA was statistically correlated with final VA (P=0.032) in the observation group. CONCLUSION: Both laser and anti-VEGF treatments are effective for hemorrhagic RAM. Combination therapy reduces the number of injections of anti-VEGF. RHA is a visual prognosis predictor in the natural history of hemorrhagic RAM.
RESUMEN
OBJECTIVES: The incidence and prevalence of inflammatory bowel disease (IBD), mainly including ulcerative colitis (UC) and Crohn's disease (CD), are increasing globally. We aimed to evaluate the potential association between IBD and nephrolithiasis, tubulointerstitial nephritis, and chronic kidney disease (CKD). METHODS: Data of hospitalized adults ≥20 years of age were extracted from the U.S. National Inpatient Sample (NIS) during 2016-2018. Patients with UC, CD, or CKD were identified through the International Classification of Diseases, Tenth Revision (ICD-10) codes. Propensity score matching (PSM) analysis (1:1) was conducted to balance the characteristics between groups. Logistic regression analyses were performed to determine the relationships between UC or CD and kidney conditions. RESULTS: Three cohorts were included for analysis after PSM analysis. Cohorts 1, 2 and 3 contained 235 262 subjects (117 631 with CD or without IBD), 140 856 subjects (70 428 with UC or without IBD), and 139 098 subjects (69 549 with CD or UC), respectively. Multivariate analysis revealed that compared to non-IBD individuals, CD patients were significantly associated with greater odds for nephrolithiasis (adjusted odds ratio [aOR] 2.25, 95% confidence interval [CI] 2.08-2.43), tubulointerstitial nephritis (aOR 1.31, 95% CI 1.24-1.38), CKD at any stage (aOR 1.28, 95% CI 1.24-1.32), and moderate-to-severe CKD (aOR 1.22, 95% CI 1.17-1.26), while UC was associated with a higher rate of nephrolithiasis. Compared to UC, CD was associated with higher odds for all such kidney conditions. CONCLUSIONS: Patients with CD are more likely to have nephrolithiasis, tubulointerstitial nephritis, CKD at any stage, and moderate-to-severe CKD compared to non-IBD individuals.
Asunto(s)
Colitis Ulcerosa , Enfermedad de Crohn , Enfermedades Inflamatorias del Intestino , Nefritis Intersticial , Nefrolitiasis , Insuficiencia Renal Crónica , Adulto , Humanos , Pacientes Internos , Puntaje de Propensión , Estudios Retrospectivos , Enfermedades Inflamatorias del Intestino/complicaciones , Colitis Ulcerosa/complicaciones , Enfermedad de Crohn/complicaciones , Enfermedad de Crohn/epidemiología , Insuficiencia Renal Crónica/etiología , Insuficiencia Renal Crónica/complicaciones , Nefrolitiasis/epidemiología , Nefrolitiasis/complicaciones , Nefritis Intersticial/epidemiología , Nefritis Intersticial/complicacionesRESUMEN
Polysaccharides are one of the most abundant and active components of Lysimachia christinae (L. christinae), which is widely adopted for attenuating abnormal cholesterol metabolism; however, its mechanism of action remains unclear. Therefore, we fed a natural polysaccharide (NP) purified from L. christinae to high-fat diet mice. These mice showed an altered gut microbiota and bile acid pool, which was characterized by significantly increased Lactobacillus murinus and unconjugated bile acids in the ileum. Oral administration of the NP reduced cholesterol and triglyceride levels and enhanced bile acid synthesis via cholesterol 7α-hydroxylase. Additionally, the effects of NP are microbiota-dependent, which was reconfirmed by fecal microbiota transplantation (FMT). Altered gut microbiota reshaped bile acid metabolism by modulating bile salt hydrolase (BSH) activity. Therefore, bsh genes were genetically engineered into Brevibacillus choshinensis, which was gavaged into mice to verify BSH function in vivo. Finally, adeno-associated-virus-2-mediated overexpression or inhibition of fibroblast growth factor 15 (FGF15) was used to explore the farnesoid X receptor-fibroblast growth factor 15 pathway in hyperlipidemic mice. We identified that the NP relieves hyperlipidemia by altering the gut microbiota, which is accompanied by the active conversion of cholesterol to bile acids.