Key candidate genes and pathways in T lymphoblastic leukemia/lymphoma identified by bioinformatics and serological analyses.

T-cell acute lymphoblastic leukemia (T-ALL)/T-cell lymphoblastic lymphoma (T-LBL) is an uncommon but highly aggressive hematological malignancy. It has high recurrence and mortality rates and is challenging to treat. This study conducted bioinformatics analyses, compared genetic expression profiles of healthy controls with patients having T-ALL/T-LBL, and verified the results through serological indicators. Data were acquired from the GSE48558 dataset from Gene Expression Omnibus (GEO). T-ALL patients and normal T cells-related differentially expressed genes (DEGs) were investigated using the online analysis tool GEO2R in GEO, identifying 78 upregulated and 130 downregulated genes. Gene Ontology (GO) and protein-protein interaction (PPI) network analyses of the top 10 DEGs showed enrichment in pathways linked to abnormal mitotic cell cycles, chromosomal instability, dysfunction of inflammatory mediators, and functional defects in T-cells, natural killer (NK) cells, and immune checkpoints. The DEGs were then validated by examining blood indices in samples obtained from patients, comparing the T-ALL/T-LBL group with the control group. Significant differences were observed in the levels of various blood components between T-ALL and T-LBL patients. These components include neutrophils, lymphocyte percentage, hemoglobin (HGB), total protein, globulin, erythropoietin (EPO) levels, thrombin time (TT), D-dimer (DD), and C-reactive protein (CRP). Additionally, there were significant differences in peripheral blood leukocyte count, absolute lymphocyte count, creatinine, cholesterol, low-density lipoprotein, folate, and thrombin times. The genes and pathways associated with T-LBL/T-ALL were identified, and peripheral blood HGB, EPO, TT, DD, and CRP were key molecular markers. This will assist the diagnosis of T-ALL/T-LBL, with applications for differential diagnosis, treatment, and prognosis.

Hematopoietic progenitor cell count as a potential quantitative marker in apheresis products during allogeneic stem cell transplantation.

BACKGROUND: The quality and quantity of hematopoietic stem cells in apheresis products are essential to the success of peripheral blood hematopoietic stem cell transplantation (PB-HSCT). While the flow cytometry measurement of CD34+ cells as a golden standard for stem cell count is labor and cost-intensive, hematopoietic progenitor cell number evaluated by XN Sysmex series automated hematology analyzers (XN-HPC) is suggested as a surrogate marker. MATERIALS AND METHODS: We evaluated the correlation and consistency of XN-HPC and CD34+ cell count in apheresis samples from both allogeneic donors and autologous patients during PB-HSCT. RESULTS: Good correlation and consistency were observed between XN-HPC and CD34+ cell counts in harvests collected from healthy donors (R = .852) rather than autologous patients (R = .375). Subgroup analysis showed that the correlation was especially poor when autologous patients used plerixafor as an additional mobilizer or were diagnosed with multiple myeloma (MM). In the setting of allogeneic transplantation, the correlation coefficients were even better in samples from non-first-round apheresis (R = .951), with high white blood cell (WBC) counts (R = .941), or having successful engraftment within 2 weeks (R = .895). ROC analysis suggested that an optimal XN-HPC count of 1127 × 106 /L best predicted a sufficient yield of CD34+ stem cells, with diagnostic sensitivity and specificity being 92% and 72%, respectively (AUC = 0.852). CONCLUSIONS: XN-HPC is a sufficient quantitative marker for stem cell assessment of harvest yield in allogeneic but not autologous HSCT.

The Heterogeneity of Ovomucoid-Specific IgE Idiotype Is Associated With Egg Allergy Symptom Severity.

Immunoglobulin E (IgE)-mediated egg allergy presents as one of the most common food allergies. The level of specific IgE (sIgE) antibody is widely used as an important in vitro diagnostic indicator. However, sIgE antibody levels are often inconsistent with the clinical manifestations of patients. The heterogeneity of egg-specific IgE idiotypes (sIgE-IDs) may help reflect clinical egg allergy severity. Eight peptides were synthesized, corresponding to the linear epitopes of ovomucoid (OVM). The sIgE-IDs of egg-allergic patients were detected by enzyme-linked immunosorbent assay. Fresh peripheral blood was collected from patients with different heterogeneity strength of sIgE-ID, and egg extract was used as a stimulus to the basophil activation test (BAT). RBL-2H3 cells were sensitized with serum with different strength of sIgE-ID heterogeneity and the release rate of ß-hexosaminidase was calculated. Among 75 patients with egg allergy, 24% had sIgE for all epitopes and 85% had sIgE for at least one epitope. Analysis of individual patients revealed differences in epitope recognition patterns among the patients, that is, heterogeneity in sIgE-ID. More importantly, the number of IgE-positive peptides had a strong correlation with allergic symptoms in egg-allergic patients (r = 0.706). BAT and RBL-2H3 cell degranulation confirmed that higher sIgE-ID heterogeneity strength was more effective in inducing effector cell responses. Our results suggest that the greater the heterogeneity strength of OVM-sIgE-ID, the more severe the allergic symptoms.

The Sysmex XN series hematopoietic progenitor cell (XN-HPC) as a predictive marker of stem cell enumeration and products: a systemic review and meta-analysis.

OBJECTIVES: Sysmex® XN series hematopoietic progenitor cell (XN-HPC) is a sensitive, fast, and economic analytical method for predicting the yields of peripheral blood stem cell enumeration and products, and does not require a sophisticated or expensive workflow. However, various studies have shown that the characteristics of its diagnostic performance were non-uniform. METHODS: We performed a systematic inquiry using PubMed, Embase, and Cochrane Library, to comprehensively search for studies published before November 21, 2021. The pooled specificity (SPE), sensitivity (SEN), negative likelihood ratio (NLR), positive likelihood ratio (PLR), diagnostic odds ratio (DOR) and receiver operating characteristic (ROC) curves were summarized to appraise the diagnostic merit of XN-HPC. A forest plot was used to research the sensitivities and specificities of XN-HPC performance. Subgroup analysis was performed to investigate heterogeneity where of importance. RESULTS: Our research included four studies that assessed the diagnostic performance of XN-HPC in hematopoietic progenitor cell collection. The pooled accuracy was 95.4% (95% CI, 94.3-96.3), SPE was 0.81 (95% CI, 0.71-0.88), SEN was 0.95 (95% CI, 0.75-0.99), NLR was 0.06 (95% CI, 0.01-0.37), PLR was 5.0 (95% CI, 3.0-8.5), DOR was 78 (95% CI, 9-707) and the summary of the area under the ROC was 0.90 (95% CI, 0.87-0.92). Forest plot of sensitivities and specificities from XN-HPC test accuracy studies indicated the existence of high heterogeneity. We deduced that the patients were the source of heterogeneity via subgroup analysis. CONCLUSIONS: XN-HPC is an excellent diagnostic marker for quantitative detection of peripheral blood hematopoietic progenitor cells.

Component resolved diagnosis of egg yolk is an indispensable part of egg allergy

Introduction and objectives It was urgent to explain the role of egg yolk allergen sensitization to the egg allergic population and we would evaluate the diagnostic value of allergen components in whole eggs, including egg white and egg yolk. Materials and methods Firstly, we collected 99 positive and 21 negative sera against egg allergy. Then we used modified enzyme linked immunosorbent assay (ELISA) to survey specific IgE (sIgE) to all-proven and single component in eggs, Ovomucoid (Gal d 1), Ovalbumin (Gal d 2), Ovotransferrin (Gal d 3), Lysozyme C (Gal d 4), Serum Albumin (Gal d 5), and YGP42(Gal d 6) in allergic and non-allergic populations. Last but not least, we studied the sIgE reactivities to egg allergen components by receiver operating characteristic (ROC) analysis. Results Among egg-allergic individuals, nearly 10% were sensitized to five of six egg allergen components, and the cross-reaction frequency between two egg yolk allergens with Gal d 1 was about 30% in the groups diagnosed with egg allergy or non-allergy. The best component-combination diagnosis in egg allergy of Gal d 1+ Gal d 6 demonstrated the largest area under curve (AUC) of 0.994. Conclusions Our results suggested that there were individual differences in allergenicity of different egg allergen components, especially in the samples negative to egg allergy diagnosed but sensitive to egg yolk components. It was indicated that component resolved diagnosis of egg yolk improved the value for egg allergy management indispensably (AU)

Establishment of a sandwich light-initiated chemiluminescence assay with double antigen for detecting human cytomegalovirus IgG antibody.

Determination of human cytomegalovirus IgG (HCMV IgG) level is of great importance in the diagnosis of HCMV infections. In this study, a novel, double antigen sandwich homogeneous immunoassay-based light-initiated chemiluminescent assay (LICA) for measuring HCMV IgG serum levels was developed. This sandwich LICA for HCMV IgG was performed by incubating serum samples with HCMV pp150 protein coated with chemibeads, streptavidin-coated sensibeads, and biotinylated HCMV pp150 protein. The working conditions of this assay were optimized and the correlation between the results of the LICA and enzyme-linked immunosorbent assay was evaluated. As a homogeneous immunoassay, this sandwich LICA could accurately and rapidly determine the serum levels of HCMV IgG with a high-throughput. Thus, this newly developed assay could be a useful analytical tool in the clinical diagnosis of HCMV infections.

Free CA125 promotes ovarian cancer cell migration and tumor metastasis by binding Mesothelin to reduce DKK1 expression and activate the SGK3/FOXO3 pathway.

Objective: CA125/MUC16 is an O-glycosylated protein that is expressed on the surfaces of ovarian epithelial cells. This molecule is a widely used tumor-associated marker for diagnosis of ovarian cancer. Recently, CA125 was shown to be involved in ovarian cancer metastasis. The purpose of this study was to investigate the mechanism of CA125 during ovarian cancer metastasis. Methods: We analyzed the Oncomine and CSIOVDB databases to determine the expression levels of DKK1 in ovarian cancer. DKK1 expression levels were upregulated or downregulated and applied with CA125 to Transwell and Western blot assays to ascertain the underlying mechanism by which CA125 stimulates cell migration via the SGK3/FOXO3 pathway. Anti-mesothelin antibodies (anti-MSLN) were used to block CA125 stimulation. Then the expression levels of DKK1were tested by enzyme-linked immunosorbent assay (ELISA) to eliminate the blocking effect of anti-MSLN to CA125 stimulation. Xenograft mouse models were used to detect the effects of CA125 and anti-MSLN on ovarian cancer metastasis in vivo. Results: DKK1 levels were downregulated in ovarian tumor tissues according to the analyses of two databases and significantly correlated with FIGO stage, grade and disease-free survival in ovarian cancer patients. DKK1 levels were downregulated by CA125 stimulation in vitro. Overexpression of DKK1 reversed the ability of exogenous CA125 to mediate cell migration by activating the SGK3/FOXO3 signaling pathway. Anti-MSLN abrogated the DKK1 reduction and increased the apoptosis of ovarian cancer cells. The use of anti-MSLN in xenograft mouse models significantly reduced tumor growth and metastasis accelerated by CA125. Conclusions: These experiments revealed that the SGK3/FOXO3 pathway was activated, wherein decreased expression of DKK1 was caused by CA125, which fuels ovarian cancer cell migration. Mesothelin is a potential therapeutic target for the treatment of ovarian cancer metastasis.

Neutrophil-to-Lymphocyte Ratio in Ovarian Cancer Patients with Low CA125 Concentration.

Ovarian cancer cases with low CA125 concentration are problematic and increase the high false negative results ratio during routine physical examination testing. Unfortunately, patients without early discovery have very low survival rates. In our study, we investigated the possible role of differential leukocyte counts and the neutrophil-to-lymphocyte ratio (NLR) in ovarian cancer patients to identify an additional discriminative marker to avoid missing diagnoses in normal physical examinations. One hundred seventy-three patients with epithelial ovarian cancer and 70 healthy controls were involved in our study. Based on the results, compared with the healthy controls, NLR was significantly different both in the low CA125 concentration group and in the complete patient group, indicating that NLR could be an effective marker for ovarian cancer screening. According to ROC, sensitivity, specificity, and NPV results, CA125 >35 U/ml is a good indicator for cancer in routine physical examination. However, in patients with low CA125 concentration, the CA125>7.65 U/ml and NLR >1.72 group yielded increased sensitivity with appropriate specificity and higher NPV results than the CA125 >35 U/ml group. We believe CA125>7.65 U/ml and NLR >1.72 should be effective makers for patients with low CA125 concentration. As a more sensitive and cost-effective strategy, this method could be conducted during routine ovarian cancer screening.

Strontium promotes osteogenic differentiation by activating autophagy via the the AMPK/mTOR signaling pathway in MC3T3­E1 cells.

Strontium (Sr) is an alkaline earth metal that exerts the dual effect of improving bone formation and suppressing bone resorption, resulting in increased bone apposition rates and bone mineral density. However, the mechanisms through which Sr exerts these beneficial effects on bone have yet to be fully elucidated. The present study aimed to reveal the underlying molecular mechanisms associated with Sr­induced osteogenic differentiation. The effects of Sr on cell proliferation and osteogenic differentiation were analyzed by MTT assay, RT­qPCR, western blot analysis, alkaline phosphatase (ALP) and Alizarin red staining assays. The extent of autophagy was determined by monodansylcadaverine (MDC) staining and western blot analysis of two markers of cellular autophagic activity, the steatosis­associated protein, sequestosome­1 (SQSTM1/p62), and the two isoforms of microtubule­associated protein 1 light chain 3 (LC3), LC­3­I/II. The expression levels of AMP­activated protein kinase (AMPK) and mammalian target of rapamycin (mTOR) were also detected by western blot analysis. Sr at a concentration of 3 mM exerted the most pronounced effect on osteogenic differentiation, without any apparent cell toxicity. At the same time, cellular autophagy was active during this process. Subsequently, autophagy was blocked by 3­methyladenine, and the enhancement of osteogenic differentiation in response to Sr was abrogated. Additionally, the phosphorylation level of AMPK was significantly increased, whereas that of mTOR was significantly decreased, in the Sr­treated group. Taken together, the findings of the present study demonstrate that Sr stimulates AMPK­activated autophagy to induce the osteogenic differentiation of MC3T3­E1 cells.

A light-initiated chemiluminescent assay for rapid quantitation of allergen-specific IgG4 in clinical samples.

BACKGROUND: An increase in allergen-specific IgG4 (sIgG4), which serves as a blocking antibody, is associated with acquisition of immune tolerance after immunotherapy. In this study, we developed a rapid, sensitive, and homogeneous immunoassay based on the light-initiated chemiluminescent assay (LICA) technology for quantifying allergen sIgG4 in serum samples. METHODS: Allergen sIgG4 was measured in vitro by incubating the sample with biotinylated allergens and chemiluminescent beads coated with anti-human IgG4 antibody, followed by the addition of streptavidin-coated sensitizer beads. Multiple tests were performed to optimize the working conditions of the LICA and evaluate its performance. RESULTS: We established the optimal concentration of biotinylated allergens (250 ng/mL), the optimal dilution range (1:8 for Gal d 1, Gal d 2 sIgG4 and 1:4 for Gal d 3, Gal d 4 sIgG4), and the optimal incubation time (20 min for Gal d 1, Gal d 2 sIgG4 and 40 min for Gal d 3, Gal d 4 sIgG4). The lower limit of quantification (LLOQ) was 0.261 ng/mL. The coefficient variation (CV) of the LICA was <10%. The assay was unaffected by general interfering substances at physiological concentrations. It exhibited excellent accuracy to detect allergen-sIgG4 in human serum. Additionally, we demonstrated that the levels of Gal d 1, Gal d 2, and Gal d 3-sIgG4 were significantly higher in the egg allergy group (p < .05), but no differences were found between the groups for Gal d 4-sIgG4. CONCLUSIONS: The LICA demonstrated satisfactory performance and can be used for quantifying allergen sIgG4 in clinical practice.