Background: Multiple Myeloma (MM) is plasma cell dyscrasia characterized by transformation of B cells into malignant cells. Although there are data regarding molecular pathology of multiple myeloma, molecularmechanisms of the disease has not been fully explained. Aims: To investigate gene expression profiles in bone marrow myeloma cells via RNA sequencing technology. Study Design: Cell study. Methods: Four untreated MM patients myeloma cells and 4 healthy donors B cells from bone marrow were sorted using FACSAria II. It was occurred two groups as patient pool from myeloma cells and the control pool from B cells. Transcriptome analysis was performed and results were analyzed using bioinformatic tools. Results: Total result of transcriptome data 18.806 transcripts (94.4%) were in pooled MM patients. 992 regions were detected as new exon candidates or altenative splicing regions. In addition, 490 mutations (deletion or insertion) and 1397 SNV were, 415 fusion transcripts, 132 frameshift mutations and 983 fusions which were reported before in NCBI were detected with unknown function were identified in patients. 35.268 transcripts were obtained (71%) (25.355 transcripts were defined previously) in the control pool. This preliminary study, the first 50 genes analyzed with a gene set enchriment analysis program by MSigDB, Enrichr and Panther.Molecular functions, cellular component, pathway and biological process of the genes obtained statistical values by means of bioinformatics tools were presented as supplemental file. Conclusion: EEF1G, ITM2C, FTL, CLPTM1L, and CYBA possible candidate genes were identified in this study which may be associated with myelomagenesis.

T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive disease resulting from the accumulation of genetic changes that affect the development of T-cells. The precise role of lymphoid enhancer-binding factor 1 (LEF1) in T-ALL has been controversial since both overexpression and inactivating LEF1 mutations have been reported to date. Here, we investigate the potential gene targets of LEF1 in the Jurkat human T-cell leukemia cell line. We used small interfering RNA (siRNA) technology to knock down LEF1 in Jurkat cells and then compared the gene expression levels in the LEF1 knockdown cells with nontargeting siRNA-transfected and non-transfected cells by employing microarray analysis. We identified DHRS2, a tumor suppressor gene, as the most significantly downregulated gene in LEF1 knockdown cells, and we further confirmed its downregulation by real-time quantitative polymerase chain reaction (qRT-PCR) in mRNA and at protein level by western blotting. Our results revealed that DHRS2 is positively regulated by LEF1 in Jurkat cells, which indicates the capability of LEF1 as a tumor suppressor and, together with previous reports, suggests that LEF1 exhibits a regulatory role in T-ALL via not only its oncogenic targets but also tumor suppressor genes.

Objectives: The role of neutrophil extracellular traps (NETs) in IgA vasculitis (IgAV) pathogenesis is emerging, with NETosis-associated markers potentially linked to disease activity. This study aimed to explore the relationship between NETosis biomarkers and IgAV disease phases. Methods: A longitudinal study involving 33 paediatric IgAV patients and 26 healthy controls was conducted. Blood and urine samples were collected from healthy controls and patients during active and inactive disease phases. NETosis markers, including cell-free DNA (cf-DNA), neutrophil elastase, MPO and citrullinated histone H3 (cit-H3) were measured using ELISA kits. Statistical analyses were conducted to compare differences for NETosis markers between groups and to evaluate correlations among variables using appropriate statistical tests. Results: There was no significant difference in gender and age between the patient and control groups. The serum cf-DNA level was significantly higher in the active patient group compared with the control and inactive patient groups (P = 0.04; P = 0.04, respectively). In urine, MPO levels were significantly lower in the active phase of patients than controls (P = 0.009), while cit-H3 levels were higher in both active and inactive phases compared with controls (P = 0.01 and P = 0.03, respectively). A cf-DNA threshold of 935 ng/ml was identified, which achieved a sensitivity of 93% (correctly identifying 93% of active patients) and a specificity of 72% (correctly identifying 72% of healthy controls). Conclusion: Elevated serum cf-DNA and urine cit-H3 suggest a potential role for NETosis in IgAV activity, highlighting these markers as potential indicators for disease monitoring. Further studies are warranted to establish standardized protocols for NETosis marker assessment in IgAV.

Resumen: Objectivos: La leucemia mieloide aguda (AML, por sus siglas en inglés) es una enfermedad muy heterogénea. Aunque se puede clasificar a los pacientes en grupos de riesgo según sus mutaciones genéticas, el pronóstico dentro de cada categoría varía sustancialmente. Es perentorio identificar nuevos marcadores moleculares de la AML. Recientemente, se ha descrito la elevación del inhibidor de la serina peptidasa Kazal tipo 2 (SPINK2) en la AML, habiendo sido relacionada con peores resultados clínicos en metaanálisis, así como en un número limitado de pacientes con AML. Métodos: Analizamos la expresión de SPINK2 en 62 pacientes (45 adultos y 17 niños) con AML y en 11 líneas celulares mediante PCR cuantitativa (qRT-PCR). Los niveles de la proteína SPINK2 se determinaron en líneas celulares mediante ELISA. Resultados: Observamos un aumento de expresión del ARNm de SPINK2 y de los niveles de la proteína en las líneas celulares de AML (HL60 y NB4), frente a otras líneas celulares (K562, Jurkat y NALM6, MCF7, HeLa, HUVEC, hFOB, 293T, U87). Los pacientes con AML mostraron una expresión elevada de ARNm de SPINK2 frente a los controles (p=0,004) y esta fue significativamente menor en los pacientes t(8;21) positivos, frente a los pacientes negativos (p=0,0006). Conclusions: Estos resultados sugieren que el gen SPINK2 tiene un papel relevante en el desarrollo de la AML. Son necesarios más estudios para evaluar la expresión de SPINK2 en los pacientes con AML con la mutación t(8.21) e investigar su valor pronóstico en varios subgrupos de pacientes con AML.

The modulatory effect of C-Vx, a novel therapeutic agent, on the immune system of COVID-19 patients was investigated. The functions of T and NK cells of COVID-19 patients with different disease severity were evaluated by flow cytometry in response to C-Vx stimulation. The levels of pro- and anti-inflammatory cytokines were detected by multiplex assay in supernatants after cell culture with C-Vx. Bradykinin, IRF3 and IFN-α levels were also measured by ELISA in the presence or absence of C-Vx stimulation. As a result, increased CD107a expression was observed on NK cells in response to C-Vx addition. The proliferation of T cell subsets was increased by C-Vx, decreasing by disease severity. IL-4 and IL-10 levels were elevated while IFN-γ and IL-17 levels were reduced in T cells following C-Vx stimulation. However, the levels of pro-inflammatory IL-1β, IL-6, IL-8, IFN-γ and GM-CSF were significantly increased upon C-Vx stimulation. IFN-α levels tended to increase after incubation with C-Vx. These findings support an immunomodulatory action of C-Vx on the immune system of patients with mild and moderate phase of COVID-19.