Mononuclear Cell Count Research Articles

Stem cell mobilizating effect of heparin in patients undergoing autologous stem cell transplantation.

Adequate stem cell collection is essential for successful stem cell transplantation. Heparin enhances stem cell mobilization by competing with heparin sulfate proteoglycans. Heparin is also used as an anticoagulant before leukapheresis. Here, we evaluated the effects of heparin on stem cell mobilization in patients who underwent autologous stem cell transplantation (ASCT). We evaluated patients who underwent ASCT. Patients were divided into two groups: those who received heparin plus citrate (heparinized patients) and those who received citrate only (nonheparinized patients) for anticoagulation. Univariate and multivariate analyses were also performed. The collection efficiency 2 (CE2) for CD34+ cells was calculated and compared between heparinized and nonheparinized patients. This study included 1017 patients. There were 478 (47%) heparinized and 539 (53%) nonheparinized patients. The number of collected CD34+ cells was significantly higher in heparinized patients (P < .00001). The multivariate analyses showed that using heparin was an independent positive factor for collected CD34+ cells (adj-R2 = 0.744; F = 369.331, P < .00001). CE2 was significantly higher in heparinized patients than in nonheparinized patients (66.8% vs 52.1%; P < .00001). The rate of collecting at least 2 × 106 /kg CD34+ cells was 3.3 times higher for heparinized patients in poor mobilizers (P < .00001). Heparinized patients had significantly higher total nucleated and mononuclear cell counts (P < .00001 and <.00001, respectively). Heparin enhances stem cell collection and increases CE2. The use of heparin may reduce the need for other strategies to increase stem cell mobilization.

African-Americans With Multiple Sclerosis Have a Higher Proportion of B-Cell Lineage Cells in Their Cerebrospinal Fluid (CSF) Than White Patients (P9-3.010)

<h3>Objective:</h3> To compare proportions of B-cell lineage CD19+ and CD20+ cells in CSF of African-Americans (AA) and White (W) patients with MS. <h3>Background:</h3> Compared to W patients, AA patients with MS are more likely to have oligoclonal bands in CSF; have higher IgG index in CSF; and have higher circulating plasmablasts in blood. It is unknown whether the proportion of B-cells in CSF differs between AA and white patients in MS. <h3>Design/Methods:</h3> Demographics, disease-related information, and treatment history were retrospectively collected on patients with neurologist-diagnosed MS who self-identified as AA or W, received care at NYU MS Care Center, and underwent flow cytometry of CSF during diagnostic work-up. Lymphocyte subsets (B-lymphocytes, T-lymphocytes, NK cells), monocytes and plasma cells were analyzed with flow cytometry (BD FACSCanto™ and FACSCanto™ II Cell Analyzers). <h3>Results:</h3> 20 AA and 57 W MS patients fulfilled inclusion criteria. The groups had similar age at lumbar puncture, sex ratio, CSF cell counts, protein and glucose CSF concentrations, and oligoclonal band number. IgG index was higher in AA compared to W (<i>1.08 v 0.87, p=0.044</i>). With respect to mononuclear cell counts in CSF, AA had higher proportions of CD19+ (<i>5.45% AA v. 2.27% W, p=0.0061</i>) and CD20+ (<i>5.00% AA v. 1.95% W, p=0.0046</i>) cells, but did not significantly differ in proportion of CD4+, CD8+, CD38+ bright B-cells, NK cells and monocytes. Sensitivity analyses that excluded all patients on disease-modifying therapy at the time of lumbar puncture (n=8) yielded similar conclusions. <h3>Conclusions:</h3> B-cells are overrepresented in the CSF of African American patients with MS relative to Whites. Whether this finding bears on the increased severity of MS disease course in AA patients warrants investigation. <b>Disclosure:</b> Mr. Xue has nothing to disclose. Dr. Arbini has nothing to disclose. Mr. Melton has received personal compensation for serving as an employee of Janssen Pharmaceuticals. Dr. Kister has received personal compensation in the range of $5,000-$9,999 for serving as a Consultant for Genentech-Roche. The institution of Dr. Kister has received research support from Genentech. Dr. Kister has received publishing royalties from a publication relating to health care.

Albumin-based solution is the ideal post-thawing suspension medium for cord blood hematopoietic stem cells: A stability and proliferative evaluation.

Cryopreservation and thawing protocols represent key factors for the efficacy of cellular therapy products, such as hematopoietic stem cells (HSCs). While the HSC cryopreservation has already been standardized, the thawing procedures have been poorly studied. This study aimed to evaluate the thawing and washing protocol of cord blood (CB) derived HSCs or the HPC(CB), by selecting the optimal thawing solution and determining CD34+ cells' stability over time. Seven cryopreserved CB products were thawed, washed, and resuspended in three different solutions (10% Dextran40 in NaCl equally prepared with 5% human albumin; 5% human albumin in PBS/EDTA; and normal saline) and stored at 4°C (±2°C). Mononuclear cell (MNC) count, CD45+/CD34+ cell enumeration, and cell viability were tested at 0, 1, 2, 4, 6, 8, 12, 24, 36, and 48 h. The protocol with the selected solution was further validated on additional 10 CB samples. The above parameters and the colony-forming unit (CFU) assay were analyzed at time points 0, 2, 4, 6, and 8 h. The results showed that the 5% human albumin was the most suitable thawing solution. MNCs were stable up to 4 h (p = 0.009), viable CD45+ cells were unstable even at 2 h (p = 0.013), and viable CD34+ cells were stable until 6 h (p = 0.019). The CFU assay proved the proliferative potential up to 8 h, although significantly decreased after 4 h (p = 0.013), and correlated with the viable CD34+ cell counts. We demonstrated that the post-thawed and washed HPC(CB) using 5% human albumin is stable for up to 4 h.

The Effect of Menstrual-Cycle Phase on Immune Responses to a 5-km Cycling Time Trial: An Exploratory Study.

Exercise has transient effects on the immune system that could influence infection risk and tissue recovery after exercise. Little is known about how the menstrual cycle interacts with the immune responses to acute exercise. This exploratory study sought to evaluate the effect of menstrual-cycle phase on peripheral blood mononuclear cell counts before and immediately after a bout of intense aerobic exercise. Seven naturally menstruating women (age: 27 [3]y) completed three 5-km cycling time trials coinciding with the early-follicular, late-follicular, and mid-luteal stage, confirmed by hormonal measurement. Venous blood samples were taken and examined for the presence of immune cell types using flow cytometry. Reductions in circulating CCR7+CD45RA+ naïve CD4+ T cells, CD4+CD25+ regulatory T cells, and CD56+CD57+ natural killer cells observed during the early-follicular phase were attenuated when exercise was performed during the late-follicular phase. Similarly, reductions in circulating CD56+CD57+ natural killer cells and CD14+TLR4+ monocytes following exercise in the early-follicular phase were abolished when exercise was performed in the midluteal phase. These preliminary findings indicate that the effect of acute high-intensity exercise on immune-cell mobilization and activation varies across the menstrual cycle, potentially impacting the anti-inflammatory effects of regulatory T cells and the cell-mediated effects of both natural killer CD57+ cells and monocytes expressing TLR4.

AAV-mediated expression of HLA-G for the prevention of experimental ocular graft vs. host disease

Ocular graft versus host disease (OGvHD) develops after allogeneic hematopoietic stem cell transplantation (HSCT) and manifests as ocular surface inflammatory disease. This study evaluated the efficacy of adeno-associated virus (AAV) gene therapy encoding human leukocyte antigen G (HLA-G) to inhibit OGvHD. A major histocompatibility mismatch chronic OGvHD murine model was evaluated. 7days after HSCT, mice were dosed subconjunctivally with scAAV8-HLA-G1/5 (1 x 109 vg/eye), topical cyclosporine (twice daily), or left untreated. Body weights and tear production (red thread test) were recorded, and eyelid, corneal opacity, and corneal fluorescein retention were scored through day 44 after HSCT. Tissues were collected for vector biodistribution, ocular histology, and immunofluorescence. Compared with untreated HSCT eyes, those dosed with scAAV8-HLA-G1/5 had significantly reduced clinical inflammatory signs of OGvHD. On histology, eyes that received scAAV8-HLA-G1/5 or cyclosporine had a significantly lower mean limbal mononuclear cell count when compared with non-treated HSCT eyes. HLA-G immunofluorescence was detected in the subconjunctiva and peripheral cornea in HSCT animals treated with scAAV8-HLA-G1/5. Vector genomes were detected in the lacrimal gland, but not in the other tested organs. These results provide evidence that subconjunctival AAV targets ocular surface and corneal disease and support that HLA-G-based gene therapy may be an effective treatment for OGvHD.

Factors Influencing the Yield of Progenitor Cells in Bone Marrow Aspiration Concentrate—A Retrospective Analysis of 58 Patients

This study aims to identify the role of subjective factors (age, sex, and comorbidities) and procedure-specific factors (aspiration volume) in influencing the yield of progenitor cells in bone marrow aspiration concentrate (BMAC) harvested from the iliac crest. A retrospective analysis was conducted on 58 patients (male:female = 31:27; mean age: 52.56 ± 18.14 years) who underwent BMAC therapy between January 2020 and June 2021. The factors analyzed include individual factors such as age, sex, and comorbid conditions, and procedural factors such as aspirate volume. The mononuclear cell (MNC) count and colony-forming unit (CFU) assay were used to assess the yield of progenitors in the aspirate. Pearson's correlation test was performed for the age, aspirate volume, and outcome parameters, such as MNC and CFU. We used the chi-square test to analyze the role of sex and comorbidities on cellular yield. The mean volume of aspirate used for BMAC therapy was 66.65 (±17.82) mL. The mean MNC count of the BMAC was 19.94 (±16.34) × 106 cells, which formed 11 (±12) CFUs. Evidence of statistically significant positive associations was noted between the CFUs developed from the BMAC and the MNC count within them (r = 0.95, p < 0.001). The sex of the individual did not play any significant role in MNC count (p = 0.092) or CFUs formed (p = 0.448). The age of the individual showed evidence of a statistically significant negative association with the MNC count (r = -0.681, p < 0.001) and CFUs (r = -0.693, p < 0.001), as did the aspiration volume with the MNC count (r = -0.740, p < 0.001) and CFUs (r = -0.629, p < 0.001). We also noted a significant reduction in the MNC count (p = 0.002) and CFUs formed (p = 0.004) when the patients presented comorbidities. Individual factors such as age, comorbid conditions, and procedure factors such as aspirate volume significantly affected the yield of progenitor cells in the BMAC. The sex of the individual did not influence the yield of progenitor cells in BMAC.

Immune Profile and Body Composition of Independent Oldest Old: The Longevous Project

Introduction: Immunosenescence is associated with changes in lymphocyte function, thymus atrophy, and a chronic inflammatory process referred to as “inflammaging,” which may in part be linked to eating disorders. Objective: The aim of the study was to determine the prevalence rate of immunological alterations, including immune risk profile (IRP), and their association with body composition in the oldest old individuals. Methods: A cross-sectional study of 201 older adults aged 80 years and over, able to walk unaided, with no cognitive or immunological impairment, and with no serious diseases was conducted. Blood samples were collected between 2012 and 2014 during the morning period, and the following tests were conducted: urea, creatinine, hemogram, fasting glucose, glycated hemoglobin, transferrin, albumin, 25-OH vitamin D, and high-sensitivity CRP. Plasma cytokines were measured and mononuclear cell counts were performed by flow cytometry. Anthropometric measurements and densitometry using dual-energy X-ray absorptiometry (DXA) were performed to assess body composition. Results: Mean age was 84.4 ± 3.7 years, and the numbers of T cells were CD4, 784.0 cell/µL; CD8, 371.0 cell/µL; and CD4/CD8, 2.4. The rate of CD4/CD8 <1 (IRP) was 9.4%, CD4/CD8 1–5 was 85.6%, while CD4/CD8 >5 was only 5.5%. CRP, tumor necrosis factor, IL1, IL4, IL6, and IL10 variables showed a high coefficient of variation but low mean values of 1.1 ± 3.1 mg/L for CRP (reference range <3 mg/L) and 3.9 ± 5.0 pg/mL for IL6 (reference range <7.0 pg/mL). The same pattern was found for all other inflammatory variables assessed, characterizing a population whose values indicated low level of inflammation, considering age. Lean mass, as measured by DXA, was higher in men than in women, while the inverse was found for fat % (p < 0.001). A positive association between CRP values and DXA fat % (p value: 0.007, r: 0.49) and a negative association between CRP values and DXA lean mass (p value: 0.046, r:–0.37) was observed. Conclusion: In the independent oldest old, IRP rate proved low and high-sensitivity CRP was shown to be associated with body composition.

Machine Learning Algorithm As a Prognostic Tool for Epstein-Barr Virus Reactivation after Haploidentical Hematopoietic Stem Cell Transplantation

AbstractBackground: Epstein-Barr virus (EBV) reactivation is one of the most important infections after haploidentical related donor (HID) hematopoietic stem cell transplantation (HSCT). We aimed to establish a comprehensive model with machine learning, which could predict EBV reactivation after HID HSCT with antithymocyte globulin (ATG) for graft-versus-host disease (GVHD) prophylaxis. Method: We enrolled 470 consecutive acute leukemia patients, 60% of them (n=282) were randomly selected as training cohort, and the remains 40% (n=188) were used as validation cohort. Results: The equation was as follows: Probability (EBV reactivation) = , where Y = 0.0250 × (age) - 0.3614 × (gender) + 0.0668 × (underlying disease) - 0.6297 × (disease status before HSCT) - 0.0726 × (DRI) - 0.0118 × (HCT-CI score) + 1.2037 × (HLA disparity) + 0.5347 × (EBV serostatus) + 0.1605 × (conditioning regimen) - 0.2270 × (donor/recipient gender matched) + 0.2304 × (donor/recipient relation) - 0.0170 × (mononuclear cell counts in graft) + 0.0395 × (CD34+ cells count in graft) - 2.4510. The threshold of probability was 0.4623, which separated patients into low- and high-risk groups. The 1-year cumulative incidence of EBV reactivation in the low- and high-risk groups was 11.0% versus 24.5% (P < 0.001), 10.7% versus 19.3% (P = 0.046), and 11.4% versus 31.6% (P = 0.001), respectively, in total, training, and validation cohorts. It also could predict relapse and survival after HID HSCT. Conclusion: We established a comprehensive model which could predict EBV reactivation in HID HSCT recipients using ATG for GVHD prophylaxis. Importance：We established a comprehensive model with machine learning in HID HSCT recipients using ATG for graft-versus-host disease prophylaxis, which separated patients into low- and high-risk groups. It could predict EBV reactivation in total, training, and validation cohort. It could predict EBV reactivation in patient with or without CMV infection, and it could also predict EBV reactivation in patients with or without acute GVHD. It could also predict relapse and survival after HID HSCT. This was the first predicted model for these patients who have a high risk of EBV reactivation and it can be popularized easily. It can also help to conduct risk stratification−directed EBV prophylaxis after HID HSCT. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

IMMU-06. REDUCTION OF CD4+ T CELLS NEGATIVELY AFFECTS SURVIVAL IN OLD BUT NOT YOUNG MICE WITH GLIOBLASTOMA

Abstract OBJECTIVE Previous work indicates poorer survival outcomes for older glioblastoma (GBM) patients ≥ 65 years of age as compared to younger patients &amp;lt; 65 years of age. The cellular mechanisms associated with this age-dependent survival difference remain elusive. Here we evaluated the overall survival of young and old immunocompetent mice with intracranial brain tumors that were depleted for specific immune cell populations. METHODS Young 6-8 weeks of age vs old 80+ weeks of age C57BL/6 mice were treated for up to 3 weeks with either IgG isotype control antibodies or with depleting antibodies against murine CD4, CD8, or NK1.1, beginning at 3 days prior to- and every 3 days post-intracranial injection of the mouse brain tumor cell lines, GL261 or CT-2A. Overall survival was compared between groups. Additionally, absolute peripheral blood mononuclear cell (PBMC) counts for CD3+ T cells, CD4+ T cells, and CD8+ T cells were analyzed between aneurysm control- and GBM-patients. RESULTS Older adult mice, but not younger mice, with either intracranial GL261 or CT-2A cell-based brain tumors have a significantly decreased overall survival when depleted for CD4+ T cells as compared to the IgG control-treated group, as well as groups depleted for CD8+ T- or NK-cells (P &amp;lt; 0.05). Coincidently, older GBM patients have fewer CD4+ T cells as compared to younger GBM patient counterparts (P &amp;lt; 0.05). CONCLUSIONS The data suggest that the levels of CD4+ T cells play more important roles in the survival of old subjects with GBM than in the younger subjects. A mechanistic understanding for this CD4+ T cell-dependent survival benefit is ongoing.

Towards defining morphologic parameters of normal parous and nulliparous breast tissues by artificial intelligence

BackgroundBreast terminal duct lobular units (TDLUs), the source of most breast cancer (BC) precursors, are shaped by age-related involution, a gradual process, and postpartum involution (PPI), a dramatic inflammatory process that restores baseline microanatomy after weaning. Dysregulated PPI is implicated in the pathogenesis of postpartum BCs. We propose that assessment of TDLUs in the postpartum period may have value in risk estimation, but characteristics of these tissues in relation to epidemiological factors are incompletely described.MethodsUsing validated Artificial Intelligence and morphometric methods, we analyzed digitized images of tissue sections of normal breast tissues stained with hematoxylin and eosin from donors ≤ 45 years from the Komen Tissue Bank (180 parous and 545 nulliparous). Metrics assessed by AI, included: TDLU count; adipose tissue fraction; mean acini count/TDLU; mean dilated acini; mean average acini area; mean “capillary” area; mean epithelial area; mean ratio of epithelial area versus intralobular stroma; mean mononuclear cell count (surrogate of immune cells); mean fat area proximate to TDLUs and TDLU area. We compared epidemiologic characteristics collected via questionnaire by parity status and race, using a Wilcoxon rank sum test or Fisher’s exact test. Histologic features were compared between nulliparous and parous women (overall and by time between last birth and donation [recent birth: ≤ 5 years versus remote birth: > 5 years]) using multivariable regression models.ResultsNormal breast tissues of parous women contained significantly higher TDLU counts and acini counts, more frequent dilated acini, higher mononuclear cell counts in TDLUs and smaller acini area per TDLU than nulliparas (all multivariable analyses p < 0.001). Differences in TDLU counts and average acini size persisted for > 5 years postpartum, whereas increases in immune cells were most marked ≤ 5 years of a birth. Relationships were suggestively modified by several other factors, including demographic and reproductive characteristics, ethanol consumption and breastfeeding duration.ConclusionsOur study identified sustained expansion of TDLU numbers and reduced average acini area among parous versus nulliparous women and notable increases in immune responses within five years following childbirth. Further, we show that quantitative characteristics of normal breast samples vary with demographic features and BC risk factors.

The characterization analysis of pathogenic T cells in immune-mediated aplastic anemia mouse model

目的通过DsRed小鼠淋巴细胞输注加全身辐照（TBI）建立免疫介导的再生障碍性贫血（AA）C.B10-H2b/LilMcd (C.B10)小鼠模型，分析C.B10 AA小鼠模型中病理性T细胞在不同器官的动态归巢过程及特征。方法通过异体淋巴细胞输注加TBI建立AA小鼠模型，以TBI组为对照，分析外周血细胞和骨髓单个核细胞数，验证造模效果。在造模的第3、6、9、12天，处死小鼠并收集骨髓、脾脏、淋巴结和胸腺单个核细胞，流式细胞术分析DsRed+ T细胞的动态变化及效应记忆T细胞（TEM）、中央记忆T细胞（TCM）、初始T细胞（naïve T）和效应T细胞（TEFF）的比例。PCR Array分析AA小鼠不同组织内DsRed+病理性T细胞活化相关基因的表达谱。结果与TBI组相比，AA组小鼠在造模第9、12天出现严重骨髓衰竭，第12天时骨髓有核细胞数、外周血细胞数均显著减少（P值均<0.05）。AA组小鼠骨髓、脾脏、淋巴结内DsRed+ T细胞比例随时间增加而增加。骨髓中的DsRed+ T细胞比例在第3、6天低于脾脏与淋巴结，但在第12天时高于脾脏与淋巴结（P值均<0.05）。整个骨髓衰竭形成过程中，胸腺中DsRed+ T细胞比例均最低。在第12天时，AA小鼠骨髓、淋巴结和脾脏单个核细胞中，DsRed+CD3+CD4+ T细胞比例分别为（91.38±2.10）％、（39.78±6.98）％、（67.87±12.77）％，三者之间两两比较均有统计学意义（P值均<0.05）；DsRed+CD3+CD8+ T细胞比例分别为（98.21±1.49）％、（94.06±4.20）％、（96.29±1.23）％，差异均无统计学意义（P值均>0.05）。第9、12天，AA组小鼠骨髓中几乎所有的DsRed+CD4+ T或DsRed+CD8+ T细胞均为TEM，而淋巴结中包括TEM、TCM以及naïve T。PCR Array结果显示：骨髓中DsRed+CD4+或DsRed+CD8+ T细胞中CD38、IFN-γ、LAG3、CSF1、SPP1及TNFSF13B表达增高，但 DsRed+CD4+ T细胞中FOXP3、CTLA4表达降低。结论DsRed小鼠淋巴细胞输注可导致C.B10小鼠发生骨髓衰竭，并可以在该AA模型小鼠中追踪DsRed+病理性T细胞。在C.B10 AA模型小鼠体内，DsRed+CD8+和DsRed+CD4+ T细胞先归巢到脾脏与淋巴结，在其中扩增分化，最终转运至骨髓并转化为TEM。胸腺对异体T细胞的归巢过程更具抵抗性，提示胸腺在免疫介导AA模型的骨髓衰竭过程中可能是一个免疫豁免部位。在C.B10 AA模型小鼠的骨髓衰竭形成过程中，归巢到骨髓的DsRed+病理性T细胞较迁移到脾脏与淋巴结的细胞具有更强的免疫活性，而免疫抑制活性则减弱。

Social Factors, Epigenomics and Lupus in African American Women (SELA) Study: protocol for an observational mechanistic study examining the interplay of multiple individual and social factors on lupus outcomes in a health disparity population

IntroductionDespite the disproportional impact of SLE on historically marginalised communities, the individual and sociocultural factors underlying these health disparities remain elusive. We report the design and methods for a study...

Serum hormone levels and normal breast histology among premenopausal women.

Breast terminal duct lobular units (TDLUs) are the main source of breast cancer (BC) precursors. Higher serum concentrations of hormones and growth factors have been linked to increased TDLU numbers and to elevated BC risk, with variable effects by menopausal status. We assessed associations of circulating factors with breast histology among premenopausal women using artificial intelligence (AI) and preliminarily tested whether parity modifies associations. Pathology AI analysis was performed on 316 digital images of H&E-stained sections of normal breast tissues from Komen Tissue Bank donors ages ≤ 45years to assess 11 quantitative metrics. Associations of circulating factors with AI metrics were assessed using regression analyses, with inclusion of interaction terms to assess effect modification. Higher prolactin levels were related to larger TDLU area (p < 0.001) and increased presence of adipose tissue proximate to TDLUs (p < 0.001), with less significant positive associations for acini counts (p = 0.012), dilated acini (p = 0.043), capillary area (p = 0.014), epithelial area (p = 0.007), and mononuclear cell counts (p = 0.017). Testosterone levels were associated with increased TDLU counts (p < 0.001), irrespective of parity, but associations differed by adipose tissue content. AI data for TDLU counts generally agreed with prior visual assessments. Among premenopausal women, serum hormone levels linked to BC risk were also associated with quantitative features of normal breast tissue. These relationships were suggestively modified by parity status and tissue composition. We conclude that the microanatomic features of normal breast tissue may represent a marker of BC risk.

The correlation between severity scores in computed tomography lung scans and viral load in the severity of novel coronavirus 2019 progression.

BackgroundThis study aimed to find the correlation between severe computed tomography (CT) lung scores and nasopharyngeal viral load (Ct value) in the severity of COVID‐19 disease progression.MethodIn this study, 37 patients diagnosed with COVID‐19 were categorized into severely ill and not severely ill samples. Their Ct values, epidemiological data, lung CT, and laboratory test results were collected three times, respectively, on the first day of their hospital admission, 3–5 days thereafter, and prior to hospital discharge. Among the 37 patients, 8 progressed from not severely ill to severely ill; we also paid attention and observed changes in clinical parameters of COVID‐19 patients who entered our city from other cities (imported cases) and the infected local residents who contacted these imported patients (non‐imported cases).ResultsAmong the 37 patients, the Ct values and lung severity scores (LSSs) were similar in imported and non‐imported cases (F = 0.59 and 2.56; p = 0.45 and 0.12, respectively) but the proportion of severely ill imported patients was significantly higher compared with non‐imported patients (F = 7.77; p = 0.01). Additionally, 21.6% of patients' illness worsened; lymphocyte counts and Ct values were significantly lowered, and C‐reactive protein and LSS significantly increased during COVID‐19 disease progression. Furthermore, LSS negatively correlated with lymphocyte and mononuclear cell counts, as well as Ct values (Pearson's rank = −0.763, −0.824, and −0.588; p = 0.028, 0.012, and 0.003, respectively).ConclusionIn the severity of COVID‐19 disease progression, nasopharyngeal viral load and lung CT severity were closely related, and LSS negatively correlated with lymphocyte and mononuclear cell counts, as well as Ct values.

MRI-based radiomics models can improve prognosis prediction for nasopharyngeal carcinoma with neoadjuvant chemotherapy

BackgroundThe purpose of this study was to explore the prognostic value of imaging features and related models in nasopharyngeal carcinoma (NPC) patients that received neoadjuvant chemotherapy. Materials and methodsWe systematically reviewed the data of 110 NPC patients who received radiotherapy and neoadjuvant chemotherapy. The patients were randomly divided into the training cohort (n = 88) and the verification cohort (n = 22). The imaging data collected in this study were screened via Pyramidics and used to construct prediction models based on histology and clinical nomographs. The models' accuracy was evaluated via calibration curves and the consistency index (C-index). In addition, we also explored the correlation between radiomics expression patterns, quantitative histological characteristics, and clinical data and then constructed a model to predict the prognosis of NPC. ResultsThe models that integrated radiomics contours with all the clinical data were superior to those based on the clinical data alone (C-index 0.746 vs. C-index 0.814, respectively) and the calibration curves showed good consistency. The heat map showed that the radiomics expression pattern and selected histological characteristics were correlated with the clinical stage, T stage, and N stage (p < 0.05), and no radiomics feature was associated with lactate dehydrogenase expression, lymphocyte count, or mononuclear cell count. ConclusionMRI-based radiomics can significantly improve the efficacy of traditional TNM staging and clinical data in predicting the progression-free survival (PFS) of patients with advanced NPC, which may provide an opportunity for precision medicine.

Identification of potential key genes and functional role of CENPF in osteosarcoma using bioinformatics and experimental analysis.

Osteosarcoma, which arises from bone tissue, is considered to be one of the most common types of cancer in children and teenagers. As the etiology of osteosarcoma has not been fully elucidated, the overall prognosis for patients is generally poor. In recent years, the development of bioinformatical technology has allowed researchers to identify numerous molecular biological characteristics associated with the prognosis of osteosarcoma using online databases. In the present study, Gene Expression Omnibus (GEO) database was used and three microarray datasets were obtained. The GEO2R web tool was utilized and differentially expressed genes (DEGs) in osteosarcoma tissue were identified. Venn analysis was performed to determine the intersection of the DEG profiles. DEGs were analyzed by Gene Ontology function and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis. Protein-protein interactions (PPIs) between these DEGs were analyzed using the Search Tool for the Retrieval of Interacting Genes database, and the PPI network was then visualized using Cytoscape software. The top ten genes were identified based on measurement of degree, density of maximum neighborhood component, maximal clique centrality and mononuclear cell counts in the PPI network, and five overlapping genes [origin recognition complex subunit 6 (ORC6), IGF-binding protein 5 (IGFBP5), minichromosome maintenance 10 replication initiation factor (MCM10), MET proto-oncogene, receptor tyrosine kinase (MET) and centromere protein F (CENPF)] were identified. Additionally, three module networks were analyzed by Molecular Complex Detection (MCODE), and six key genes [ORC6, MCM10, DEP domain containing 1 (DEPDC1), CENPF, TIMELESS interacting protein (TIPIN) and shugoshin 1 (SGOL1)] were screened. Combined with the results from Cytoscape and MCODE, eight hub genes (ORC6, MCM10, DEPDC1, CENPF, TIPIN, SGOL1, MET and IGFBP5) were obtained. Furthermore, Kaplan-Meier plotter survival analysis was used to evaluate the prognostic value of these eight hub genes in patients with osteosarcoma. Oncomine and GEPIA databases were applied to further confirm the expression levels of hub genes in tissue. Finally, the functional roles of the core gene CENPF were investigated using Cell Counting Kit-8, wound healing and Transwell assays, which indicated that CENPF knockdown inhibited the proliferation, migration and invasion of osteosarcoma cells. These results provided potential prognostic markers, as well as a basis for further investigation of the mechanism underlying osteosarcoma.

Performance evaluation of automated cell counts compared with reference methods for body fluid analysis.

Cellular analysis of body fluids (BFs) can assist clinicians for the diagnosis of many medical conditions. The aim of this work is the evaluation of the analytical performance of the UF-5000 body fluid mode (UF-BF) analyzer compared to the gold standard method (optical microscopy, OM) and to XN-1000 (XN-BF), another analyzer produced by the same manufacturer (Sysmex) and with a similar technology for BF analysis. One hundred BF samples collected in K3EDTA tubes were analyzed by UF-BF, XN-BF and OM. The agreement was evaluated using Passing and Bablok regression and Bland-Altman plot analysis. The receiver operating characteristic (ROC) curves were selected for evaluating the diagnostic agreement between OM classification and UF-BF parameters. Comparison between UF-BF and OM, in all BF types, showed Passing and Bablok's slope comprised between 0.99 (polymorphonuclear cells count, PMN-BF) and 1.39 (mononuclear cells count, MN-BF), the intercepts ranged between 26.47 (PMN-BF parameter) and 226.80 (white blood cell count). Bland-Altman bias was comprised between 7.3% (total cell count, TC-BF) and 52.9% (MN-BF). Comparison between UF-BF and XN-BF in all BF showed slopes ranged between 1.07 (TC-BF and PMN-BF) and 1.16 (MN-BF), intercepts ranged between 8.30 (PMN) and 64.78 (WBC-BF). Bland-Altman bias ranged between 5.8 (TC-BF) and 21.1% (MN-BF). The ROC curve analysis showed an area under the curve ranged between 0.9664 and 1.000. UF-BF shows very good performance for the differential counts of cells in ascitic, pleural and synovial fluids and therefore it is useful to screen and count cells in this type of BF.

Clinical Characteristics of Acute Myeloid Leukemia Patients with RUNX1 Gene Mutation

To investigate the incidence of Runt-related transcription factor 1 (RUNX1) gene and its associated gene mutations in patients with acute myeloid leukemia (AML), and analyze its clinical characteristics and prognosis. The genomic DNA-PCR method was used to detect the exon of RUNX1 gene, and the gene mutations were analyzed by genetic sequencing. NPM1, DNMT3A, FLT3-ITD, IDH1/2, K/N-RAS, CEPBA, TET2, and WT1 co-mutations were also detected. Patients were followed up to determine efficacy and prognosis. Among 171 patients, the RUNX1 gene mutation was detected in 17 cases, and the mutation rate was 9.9%. The type of RUNX1 gene mutation was 9 missense mutations, 4 frameshift mutations, and 4 nonsense mutations. The peripheral blood leukocyte count of the patients in mutation group was 3 (1-101) ×109/L, which was significantly lower than those in the non-mutation group [26 (1-298)×109/L] (P=0.002), while the platelet count was 79 (22-166)×109/L, which was higher than 50 (8-351)×109/L in the non-mutation group (P=0.010), and the proportion of bone marrow blasts of the patients in the mutation group was 37 (0-72)%, which was lower than 53 (0-98)% in the non-mutation group (P=0.020). The RUNX1 mutation rate in M0 type was 55.6%, and in M4 type was 13.6%, which was significantly higher than other FAB subtypes (P=0.003). There was no significant difference in age, sex, hemoglobin concentration, and counts of peripheral blood mononuclear cells (P>0.05). The prognosis of cytogenetics in the patients in the middle and high-risk groups was 88.1% and 89.7%, which were significantly higher than that in the low-risk group, and the difference showed statistically significant (P=0.018). There was no significantly relationship between RUNX1 and specific karyotype abnormalities, including Trisomy 8, Del (7q), t (8; 21), and Inv (16) (P>0.05). There was a significant relationship between RUNX1 gene mutation and IDH1/2, N/K-RAS gene mutation (P<0.01). The complete response rate (CR) of the patients with chemotherapy in the RUNX1 mutation group(37.5%) was significantly lower than 79.4% in the non-mutated group (P=0.001). The overall survival (OS) of the patients in RUNX1 gene mutation group was lower than that in non-mutation group (P<0.05). AML patients with RUNX1 gene mutation shows unique clinical and biological characteristics, RUNX1 mutation can be regarded as a molecular marker of poor prognosis in AML patients.

232 Immunomodulation with AM3 and antioxidants creates an adequate framework for skin repair and decreases the monocyte proinflammatory stage in smoker women

Tobacco smoke has been considered a crucial factor in promoting skin and systemic aging and is also associated with the development of a low-level, systemic, chronic inflammation known as “inflammaging” where monocytes play a pivotal role. In our study, 34 healthy smoker middle- aged women were 1:1 randomly assigned to AM3 plus antioxidants and placebo treatment for three months. Peripheral mononuclear blood cells count, and cutaneous biopsy were obtained before and after the treatment and flow cytometry and immunohystologycal studies, respectively, were performed. AM3 is a glucomannan of natural origin from Candida utilis cell wall. AM3 plus antioxidants treatment significantly reduced the monocyte production of the proinflammatory IL-1, TNFα and IL-6 cytokines and also increased the regulatory IL-10 in middle-aged smoker women. The active combination also stimulated a significative increase in reticular dermis depth as well as an increase in the expression of CD117 and CD31. CD117 (also known as c-kit) is a tyrosine kinase receptor that is found in mast cells and whose expression may correlate with the increase in reticular dermis depth due to the production of cytokines such as TGFβ. As seen in fibroblasts, TGFβ-1 and TGFβ-2 are potent enhancers of elastin gene expression. On the other hand, CD31 is a glycoprotein present on the surface of endothelial cells. The increase in the expression of CD31 is related with an enhancement in dermis vascularization. In conclusion, AM3 and antioxidants treatment reduces the systemic proinflammatory monocyte disturbance of heathy smoker middle- aged women and favors the generation of cutaneous repair mechanisms.

Cerebrospinal fluid markers in incident pediatric-onset multiple sclerosis: a nationwide study

To investigate whether cerebrospinal fluid (CSF) markers differ between pediatric-onset multiple sclerosis (PoMS, onset < 18 years) and adult-onset (AoMS), and whether these markers are associated with clinical outcomes among PoMS. Prospective nationwide registry study of incident MS, including persons with a CSF sample < 3 years post-MS onset. We compared CSF oligoclonal band (OCB) status, immunoglobulin G (IgG) index levels, and mononuclear cell count between PoMS and AoMS. Within the PoMS cohort we analyzed the association between CSF markers, relapse rate and Expanded Disability Status Scale (EDSS) score, using negative binomial regression and generalized estimating equations, respectively. The cohort consisted of 130 PoMS and 3228 AoMS cases. The PoMS group had higher odds of OCB-positivity (odds ratio: 2.70; 95% CI 1.21–7.67). None of the CSF markers were associated with relapse rate in the PoMS cohort; however, OCB-positivity was associated with higher EDSS scores. This study suggested that PoMS more commonly display CSF evidence for intrathecal IgG production than AoMS. Further, we found evidence of a relationship between OCB-positivity and subsequent disability, suggesting that they could play a role in the prognostication of MS in children.