miR-539-5p targets BMP2 to regulate Treg activation in B-cell acute lymphoblastic leukemia through TGF-ß/Smads/MAPK.

MicroRNAs (mRNAs) were believed to play an important role in cancers, and this study aimed to explore the mechanism of miRNA regulating Treg in B-cell acute lymphoblastic leukemia (B-ALL). Firstly, the differentially expressed miRNAs and target genes significantly associated with Tregs were screened out by high-throughput sequencing, and their enrichment pathways were analyzed. The binding relationship between miRNA and target genes was further verified, and the effects of miRNA on the proliferation and apoptosis of B-ALL Nalm-6 cells and Treg activation were analyzed. Results showed that differentially expressed miR-539-5p was significantly under-expressed, and its target gene BMP2 was significantly over-expressed in B-ALL, and significantly enriched in the TGF-ß1 pathway. In addition, both miR-539-5p and BMP2 were significantly correlated with Treg activity in B-ALL. In vitro experiments further confirmed that miR-539-5p could directly target BMP2. The low expression of miR-539-5p in B-ALL significantly promoted BMP2 expression to promote the proliferation and inhibit apoptosis of Nalm-6 cells. Furthermore, the high expression of BMP2 in B-ALL could cooperate with TGF-ß1 to promote the activation of human CD4+CD25-T cells to Treg, and significantly activate the TGF-ß/Smads/MAPK pathway. In vivo experiments also confirmed that overexpression of miR-539-5p significantly inhibited BMP2 to suppress Treg activation and Smad1 and Smad2 phosphorylation, and finally inhibit the B-ALL process. In conclusion, miR-539-5p was significantly under-expressed in B-ALL and could target BMP2 to promote its expression, and the overexpressed BMP2 further promoted Treg activation in B-ALL by regulating TGF-ß/Smads/MAPK pathway.

Current treatment strategies targeting histone deacetylase inhibitors in acute lymphocytic leukemia: a systematic review.

Acute lymphocytic leukemia is a hematological malignancy that primarily affects children. Long-term chemotherapy is effective, but always causes different toxic side effects. With the application of a chemotherapy-free treatment strategy, we intend to demonstrate the most recent results of using one type of epigenetic drug, histone deacetylase inhibitors, in ALL and to provide preclinical evidence for further clinical trials. In this review, we found that panobinostat (LBH589) showed positive outcomes as a monotherapy, whereas vorinostat (SAHA) was a better choice for combinatorial use. Preclinical research has identified chidamide as a potential agent for investigation in more clinical trials in the future. In conclusion, histone deacetylase inhibitors play a significant role in the chemotherapy-free landscape in cancer treatment, particularly in acute lymphocytic leukemia.

The possibility of automatic capillary blood testing in routine blood tests: an evaluation of the automatic mode of the Mindray BC-7500 CRP Auto Hematology Analyzer for capillary blood testing.

Background: Capillary blood is a common specimen type used for infant blood routine tests. Until now, this specimen type could only be tested with the manual mode in hematology analyzers. Manual sample mixing and loading increases the amount labor force and can be more easily affected by human factors. This study was designed to investigate the proficiency of the automatic mode of the Mindray BC-7500 CRP Auto Hematology Analyzer for capillary blood testing. Methods: The complete blood count (CBC) results for capillary blood were compared between the automatic and manual modes. Special types of samples, including samples with high or low volume, thalassemia red cells, high fibrinogen, high hematocrit (HCT), or high triglyceride levels, were compared and evaluated. The intraclass correlation coefficient (ICC) was used to define the agreement between the 2 modes. The industry standard Analytical Quality Specifications for Routine Tests in Clinical Hematology (WS/T 406-2012), published by the National Health Commission of China, was used to evaluate the correlation between the results from the 2 modes. Results: There was good correlation between the automatic and manual modes for every type of sample, and the ICCs were all higher than 0.9. Except for high HCT or high triglyceride samples, there were no differences found between the 2 modes based on the WS/T 406-2012 standard. Conclusions: This new automatic mode utilized in the Mindray BC-7500 CRP Auto Hematology Analyzer for capillary blood yielded the same results as the manual mode except in the case of samples with high HCT or triglycerides. Capillary blood might be routinely tested automatically with hematology analyzers in the near future, which might reduce the labor required and improve standardization.

Cytokine network imbalance in children with B-cell acute lymphoblastic leukemia at diagnosis.

Immune imbalance has been proved to be involved in the pathogenesis of hematologic neoplasm. However, little research has been reported altered cytokine network in childhood B-cell acute lymphoblastic leukemia (B-ALL) at diagnosis. Our study aimed to evaluate the cytokine network in peripheral blood of newly diagnosed pediatric patients with B-ALL. Serum levels of interleukin (IL)-2, IL-4, IL-6, IL-10, tumor necrosis factor (TNF), interferon (IFN)-γ, and IL-17A in 45 children with B-ALL and 37 healthy control children were measured by cytometric bead array, while the level of transforming growth factor-ß1 (TGF-ß1) in the serum was measured by enzyme-linked immunosorbent assay. Patients showed a significant increase in IL-6 (p < 0.001), IL-10 (p < 0.001), IFN-γ (p = 0.023) and a significant reduction in TGF-ß1 (p = 0.001). The levels of IL-2, IL-4, TNF and IL-17A were similar in the two groups. Higher concentrations of pro-inflammatory cytokines were associated with febrile in patients without apparent infection by using unsupervised machine learning algorithms. In conclusion, our results indicated a critical role for aberrant cytokine expression profiles in the progression of childhood B-ALL. Distinct cytokine subgroups with different clinical features and immune response have been identified in patients with B-ALL at the time of diagnosis.

Establishment of Review Criteria Coordinating With the Automated Digital Cell Morphology Identification System in a Specialized Women's and Children's Hospital.

OBJECTIVE: We aimed to establish appropriate review criteria for blood cell analysis in a specialized women's and children's hospital. Also, the CellaVision DI-60, was developed as one of the automated digital cell morphology analyzer, we evaluated if it was shown to be most effective under the certain review criteria. METHODS: A total of 2890 blood samples were detected to optimize the previously established review criteria for women and children with the Sysmex XE-2100. A total of 623 samples were used to validate the criteria. RESULTS: The microscopic-review rate based on the initial review criteria was 51.0%. After optimization, it was reduced to 17.3% and the false-negative rate was 3.85%. There was > 80% consistency between manual review results and CellaVision DI-60 preclassification when samples triggered the platelet- or red cell-related rules. The sensitivity for abnormalities (immature granulocytes, nucleated red blood cells) of reclassification was 90% to 100% and the false-negative rate was < 5%. However, direct microscopic review was required when the "Blasts/AbnLympho?" and "Atypical Lympho?" flags were triggered. CONCLUSION: Specialized review criteria are needed for women and children. An automated morphology identification system might help to improve the review criteria.

Prognostic Relevance of Structural Maintenance of Chromosomes 4 Overexpression in Pediatric Acute Lymphoblastic Leukemia.

BACKGROUND: Acute lymphoblastic leukemia (ALL) is the most common cancer in children. Abnormal expression of structural maintenance of chromosomes (SMC) 4 has been observed in multiple tumors and plays a vital role in cancer development. However, the association between SMC4 expression and clinical characteristics in Chinese childhood patients with ALL is unknown. Thus, this study aimed to investigate the relationship between SMC4 expression and clinical features and prognosis in these patients. METHODS: Real-time quantitative polymerase chain reaction (PCR) was performed to detect the expression of SMC4 in Chinese pediatric ALL patients and in patients achieving complete remission (CR). Then, the relationships between SMC4 expression and clinical features, such as gender, age, white blood cell (WBC) count, French-American-British (FAB) classification, immunophenotype, fusion gene, prednisone response, and minimal residual disease (MRD) were determined. Furthermore, survival and prognostic factor analyses were carried out to examine the prognostic value of SMC4 expression. RESULTS: The expression level of SMC4 was significantly higher in bone morrow cells of newly diagnosed pediatric ALL patients than in those of healthy controls (p = 0.006), especially in B-cell precursor ALL (BCP ALL). Moreover, SMC4 expression was correlated with different clinical parameters. Furthermore, a decrease of SMC4 expression was detected in BCP ALL patients achieving CR. The high SMC4 expression group had both worse event-free survival rate and poorer overall survival rate. However, multivariate analysis showed that SMC4 expression was not an independently predictive factor of BCP ALL outcome. CONCLUSIONS: These results revealed that SMC4 expression in BM was associated with various clinical outcomes in pediatric patients with ALL, although it was not an independent outcome factor.

Longer Time Intervals From Symptom Onset to Diagnosis Affect the Overall Survival in Children With Acute Lymphoblastic Leukemia.

BACKGROUND: Acute lymphoblastic leukemia (ALL) is the most common childhood cancer. Early diagnosis and timely treatment are essential for effective cancer control and have been widely analyzed in childhood cancer. However, few studies have described the time to diagnosis and treatment in children with ALL. This study investigated delays in diagnosis and treatment initiation and their impact on survival. METHODS: This retrospective cohort study included 419 patients 0 to 14 years old at a tertiary hospital between 2011 and 2015. The optimal cutoff values for delays were determined by X-tile software. The Kaplan-Meier method and Cox regression models were used to evaluate the impact of delays on survival. RESULTS: The median diagnosis, treatment, and total delays were 21 (interquartile range [IQR]: 11-35), 4 (IQR: 2-7), and 26 (IQR: 16-43) days, respectively. The results of multivariate analyses showed that diagnosis delay, risk stratification, and minimal residual disease level were independent predictors for treatment outcome in childhood ALL. CONCLUSIONS: These findings suggested that a longer time to diagnosis negatively affected the clinical outcome of childhood ALL. Reducing the time to diagnosis could help to improve survival in these patients.

How to Achieve Standardization? Diluted Russell Viper Venom Test for Lupus Anticoagulant Detection in a Chinese Female Population.

On an international scale, guidelines and proposals for lupus anticoagulant detection have been published over the last 20 years, but until now, standardization has not been completely realized. The aim of this study was to evaluate the different ways of interpreting the results of lupus anticoagulant detection for standardization. A retrospective review of 15 447 instances of lupus anticoagulant detection by the diluted Russell Viper Venom test for female patients presenting with problems relating to the areas of reproduction, gynecology and obstetrics was performed. Lupus anticoagulant data were compared between different departments, months, reagent lots and cutoffs. Significant differences were found in patient data between different reagent lots, especially between lots of screening reagents (monthly average: highest 37.96 s vs lowest 33.88 s) and in the positive rates of lupus anticoagulant by different detection cutoffs (47.58% by using LA1/LA2 > 1.20 without normalization as a cutoff in Lot 1 vs 1.52% by using LA1 > 44 s as a cutoff in Lot 3). Compared with the cutoff using the value above the 99th percentile of LA1 for the healthy donors per lot, the cutoff using integrated tests with normalization had the smaller deviation of positive rate between different reagent lots. Pregnant women had higher LA1/LA2 levels than nonpregnant women. Based on the results, normalization is needed because there are significant lot-to-lot variations. Integrated tests with normalization might be a better standard by which to confirm lupus anticoagulant. Pregnant women should have population-specific cutoffs because they have higher LA1/LA2 levels.

Combined use of peripheral blood blast count and platelet count during and after induction therapy to predict prognosis in children with acute lymphoblastic leukemia.

ABSTRACT: Several studies have reported an association between the rapidity of reduction in peripheral blood blast count or recovery of normal hematopoiesis and treatment outcome during therapy in children with acute lymphoblastic leukemia (ALL). However, little is known about the impact of both of these aspects on prognosis in pediatric ALL. Accordingly, the purpose of this study was to evaluate whether the combined use of blood blast count and platelet count could predict event-free survival (EFS) and overall survival (OS) when minimal residual disease (MRD) detection was not available.A total of 419 patients aged 0 to 14 years diagnosed and treated for ALL between 2011 and 2015 were enrolled.Patients with a blast count ≥0.1 × 109/L on day 8 exhibited significantly lower survival rates than that in those with blast counts <0.1 × 109/L. The EFS and OS in patients with platelet count ≥100 × 109/L on day 33 were significantly higher than those with platelet counts <100 × 109/L. In univariate and multivariate analyses, patients with low blast count on day 8 and high platelet count on day 33 were significantly associated with better EFS and OS. The combination of blast cell count on day 8 and platelet count on day 33 demonstrated a strong association with MRD-based risk stratification.Complete blood count is an inexpensive, easy to perform, and reliable measurement in children with ALL. The combination of blast count and platelet count during and after induction chemotherapy was a significant and independent prognostic factor for treatment outcome in pediatric ALL.

Clinical features and outcome of pediatric acute lymphoblastic leukemia with low peripheral blood blast cell count at diagnosis.

ABSTRACT: Peripheral blood (PB) blast cell count on day 8 of prednisone therapy has been considered one of the strongest predictors of outcome in children with acute lymphoblastic leukemia (ALL). However, little is known about the clinical features and prognostic impact of PB blast cell count at diagnosis in these patients. The aim of this study was to evaluate the relationship between initial PB blast cell count and clinical prognosis of pediatric ALL.The study comprised 367 patients with ALL, aged 0 to 14 years, enrolled and treated using the Chinese Children's Leukemia Group-ALL 2008 protocol between 2011 and 2015. The majority (91.6%) of patients were B-cell precursor ALL (BCP ALL), and 8.4% were T-cell ALL (T-ALL).Patients with BCP ALL in the low PB blast cell count group (<1 × 109/L) had significantly superior survival rates to those in the high count group (≥30 × 109/L). In T-ALL, the low count group showed significantly inferior survival rates compared to both the intermediate count group (1-29.9 × 109/L) and high count group. Multivariate analysis revealed that the initial white blood cell count and minimal residual disease at the end of induction therapy were independently predictive of BCP ALL outcome, while risk stratification was shown to be an independent prognostic factor for T-ALL outcome.These results indicated that low blast cell count in PB at diagnosis was associated with different clinical outcomes in patients with BCP ALL and T-ALL, although it was not an independent outcome predictor by multivariate analysis.

Clinical value of the quantitation of average daily platelet increase during the recovery period in childhood acute lymphoblastic leukaemia.

The time to platelet recovery (TPR) is becoming a predicting factor during the treatment of childhood acute leukaemia. However, the initial pre-treatment platelet count (PPC) could interfere with TPR. Here, we integrated both TPR and PPC as the average daily platelet amount increase (Ap) to predict the prognosis in childhood B-ALL during the recovery period.148 patients were enrolled. The relationship between the Ap and MRD was evaluated, and Multivariate analysis was performed to evaluate whether Ap was independently associated with a better EFS. The PPC was inversely correlated with TPR (rs = -0.519, P = 0.021). Patients in Ap >3.9 × 109/L group had better EFS (x2 = 3.109, P = 0.028) than TPR ≤ 16d. Multivariate analysis indicated that Ap > 3.9 × 109/L was independently associated with a longer EFS (RR = 3.468; 95%CI: 1.037-11.597, P = 0.043). However, when introducing both MRD and Ap > 3.9 × 109/L as candidate variables, the Ap > 3.9 × 109/L lost its independent effect (P = 0.081). The strong association between MRD on treatment day 33 and Ap > 3.9 × 109/L (x2 = 148.00, P = 0.000) was responsible for this phenomenon. Ap could be a valuable prognostic indicator in childhood B-ALL.

Comparison between capillary zone electrophoresis and capillary isoelectric focusing for thalassemia screening in southern China.

BACKGROUND: Capillary isoelectric focusing is a type of capillary electrophoresis method newly used for thalassemia screening in China. Although the good performance has been proved by several studies, whether it can best suit the special needs of Chinese patients still requires further investigations. METHODS: Comparisons were made between capillary zone electrophoresis method applied on Sebia Minicap and capillary isoelectric focusing method applied on Helena V8 platform E-class on identifying Hb E, Hb CS, Hb H, and Hb Barts for patients from southern China. And mixing studies were used to evaluate the lowest detection limits of these 2 kinds of capillary electrophoresis system. RESULTS: Helena V8 could not make a distinction between peaks of Hb E and peaks of Hb A2 as Sebia Minicap did. All chosen patients with Hb H and/or Hb Barts could be screened out by both 2 systems, but when analyzed by Helena V8, it was hard to distinguish Hb H from Hb Barts sometimes, while Sebia Minicap could make a clear distinction between peaks of Hb H and Hb Barts. Only a part of patients (3 of 8, 37.5%) with Hb CS could be screened out by Helena V8, while all patients could be identified by Sebia Minicap. Sebia Minicap had a lower detection limit for trace peaks than Helena V8 (near to 0.2% vs near to 0.6%). CONCLUSIONS: Compared with capillary zone electrophoresis, capillary isoelectric focusing applied on Helena V8 maybe is not the first choice for hemoglobinopathy testing in southern China.

No prognostic significance of immunophenotypic changes at the end of remission induction therapy in children with B-lineage acute lymphoblastic leukemia.

Detection of aberrant antigen expression in acute lymphoblastic leukemia (ALL) by flow cytometric is proposed for the quantification of minimal residual disease (MRD). There are few studies that investigate the stability of the antigen expression in children with B lineage ALL at the end of remission induction therapy and determine its prognostic impact. Between 2010 and 2015, 691 bone marrow specimens of childhood ALL were sent at diagnosis for immunophenotypic characterization, and follow-up samples for MRD were analyzed on day 33. Of these, 155 patients with MRD more than or equal to 0.01% were eligible for the study. Immunophenotypic studies were performed by multiparametric flow cytometry using four-colour monoclonal antibody combinations. Overall, 86 of 155 (55.5%) cases showed phenotype shifts at least one marker. CD19 was the most stable markers. By contrast, CD20 was significantly different between diagnosis and day 33 in nearly one third of the cases. Shifts of antigen expression was not significantly associated with EFS, RFS or OS (P > 0.05). Multivariate analysis showed that WBC and BCR-ABL have independent prognostic value in childhood ALL. Changes in antigen expressions were commonly occurred at the end of induction and not associated with prognostic value in patients whose MRD were positive on day 33.

Association of 25-hydroxyvitamin D status with obesity as well as blood glucose and lipid concentrations in children and adolescents in China.

BACKGROUND: Vitamin D is an essential micronutrient during the growth and development of the human body. Previously, most of the researches about the relationship of vitamin D and obesity were focused on adults. In this paper, the relationship between serum 25-hydroxyvitamin D [25(OH)D] and obesity among children and adolescents in China was analyzed. METHODS: From August 2012 to August 2014, 443 children and adolescents participated in this study. Serum 25(OH)D, lipid and glucose were measured in the laboratory. All the participants were classified into 2 groups (obese and non-obese group) on the basis of their BMI. They were then categorized into vitamin D deficiency, vitamin D insufficiency and vitamin D normal groups according to the serum concentrations of 25(OH)D. RESULTS: The concentration of serum 25(OH)D in the obese group was significantly lower than that of the non-obese group (P<0.001). The concentration of total cholesterol, low-density lipoprotein cholesterol, fasting blood glucose, fasting insulin and homeostasis model assessment for insulin resistance showed significant differences between the obese and non-obese groups (P<0.05). CONCLUSIONS: Children and adolescents suffering from obesity are more likely to have lower concentrations of serum 25(OH)D.

Evaluation of the automated urine particle analyzer UF-1000i screening for urinary tract infection in nonpregnant women.

BACKGROUND: Urinary tract infection (UTI) is a widespread disease in women. Urine culture is still the "gold standard" diagnostic test for UTI, but most of them are negative. To reduce unnecessary culture, we evaluated the automated urine particle analyzer UF-1000i screening for UTI in nonpregnant women. METHODS: The urine specimens submitted to our laboratory were submitted for culture and tested by the Sysmex UF-1000i. Bacteria and white blood cell (WBC) counts were compared to standard urine culture results to assess the best cutoff values. RESULTS: In this study, 272 urine samples were included, of which 98 (36.0%) were culture positive with a bacterial cutoff value of 10 x 10(5) CFU/mL. A combination of bacterial (> 95/microL) and/or WBC count (> 24/microL) provided the best screening for UTI, with a sensitivity of 0.99 and a specificity of 0.82 compared with the urine culture. CONCLUSIONS: Sysmex UF-1000i could be used as a screening test for UTI in nonpregnant women. According to the distribution and range of the bacterial scattergram, we could primarily identify and differentiate between Gram-negative and Gram-positive bacteria.

Proficiency testing experience with College of American Pathologists' programs at a university hospital in China from 2007 to 2011.

CONTEXT: The first-class hospital laboratories are required to participate in a proficiency testing (PT) program by the Ministry of Health of the People's Republic of China. The College of American Pathologists (CAP) PT programs have increasingly become laboratories' preferred choices because of the well-prepared specimens, more comprehensive programs, scientific evaluation, and useful educational opportunities involved. OBJECTIVE: To evaluate the PT performance of our laboratory from 2007 to 2011 through the selected analytes and tests. DESIGN: The PT results of 22 commonly performed tests in 15 events were evaluated. The rates of unacceptable results for all analytes and tests during the 5-year study period were compared with χ (2) test. Reasons for all unacceptable results were sorted into 8 groups. RESULTS: A total of 13 of 22 analytes and tests (59.1%) achieved full scores (100%), whereas 6 analytes and tests failed to reach the goal of getting a score of 80% or higher in PT. Seen from the relative distance of the results from the target as a percentage of allowed deviation, the performance of some analytes was excellent, including albumin, amylase, lactate dehydrogenase, potassium, protein, total, uric acid, red blood cell, white blood cell, and hemoglobin. Rates of unacceptable results for all analytes demonstrated a trend of decline. The top 3 reasons for unacceptable results were identification errors in morphologic tests, specimen problems, and technical problems (25.74%, 23.76%, and 14.85%, respectively). CONCLUSIONS: The PT performance demonstrated a trend of improvement from 2007 to 2011. Proficiency testing contributed to the improvement of laboratory performance, especially the promotion of better patient care.

Two cases of false platelet clumps flagged by the automated hematology analyzer Sysmex XE-2100.

BACKGROUND: Automated hematology analyzer is widely used by clinical laboratories to examine blood cell counts and differential leukocyte counts. However, a microscopic examination of a well-prepared blood film is still necessary and useful in most cases. METHODS: Two patients with severe disease were required complete blood counts (CBC) after admission at West China Second University Hospital in Southwest China. RESULTS: A false "platelet clumps?" flag was generated by the automated hematology analyzer Sysmex XE-2100. However, when the well-prepared blood film was reviewed, there was echinocytosis rather than clumps of platelets. In addition, both patients had severe metabolic acidosis with arterial blood pH <7.00 and HCO3- <4 mmol/l. On day 2 of admission, the false platelet clumps flags were not presented in both cases. CONCLUSIONS: Our further results suggested that the false platelet clumps had been flagged probably due to the influence of the extremely low blood pH. These two cases emphasized that a microscopic examination of a blood film was central to the morphology of blood cells. When the result of microscopic examination was abnormal, it was suggested that the physician should consider further testing to find underlying condition.

One-year follow-up of patients with melamine-induced urolithiasis in Southwest China.

In 2008, more than 290,000 children suffered from urinary tract stones with a history of consuming melamine-contaminated powdered formula in China. Little was known about the long-term follow-up of outcomes of these patients in southwest China, so we conducted one-year follow-up investigation for further study. Thirty-six of 51 inpatients with melamine-induced urolithiasis were enrolled in this study. After 12 months follow-up, none of the children had vomiting, oliguria, gross hematuria, or fever. Twenty-seven patients were stone free, stones decreased in size in six patients and increased in three. Serum total protein, albumin, and pre-albumin increased significantly from baseline to 12 months, and renal function remained normal. Melamine-induced urolithiasis could not lead renal dysfunction at 12 months follow-up. The results of our study indicated that more attention should be paid to the prevention of protein malnutrition in children with a history of consuming melamine-contaminated food products.