A sample-preparation-free, automated, sample-to-answer system for cell counting in human body fluids.

While many clinical laboratory tests are now highly automated, body fluid cell counting, particularly in low-cellularity samples such as cerebral spinal fluid (CSF), is often performed manually. Here, we report a simple, cost-effective method to obtain white and red blood cell counts from human body fluids such as CSF. The method consists of a compact, automated, and low-cost fluorescence microscope system, coupled to a sample chamber containing all of the necessary reagents in dry form to stain and prepare the sample. Sample focus and scanning are handled automatically, and the acquired multimodal images are automatically analyzed to extract cell counts. Comparison with manual counting on over 200 clinical samples shows excellent agreement. As the system counts a substantially larger image region than a standard manual cell count, we find our sensitivity to extremely low cellularity samples to potentially be higher than the manual gold standard, evidenced by our system recording images of cells in samples whose cell count was registered as "0" by a trained user. Thus, our system holds promise for routine, automated, and sensitive analysis of body fluids whose cellularity extends across a wide dynamic range.

Cytogenetic and mutational analysis and outcome assessment of a cohort of 284 children with de novo acute myeloid leukemia reveal complex karyotype as an adverse risk factor for inferior survival.

BACKGROUND: Acute myeloid leukemia (AML) is rare in children. Although complex karyotype (CK) defined as ≥ 3 cytogenetic abnormalities is an adverse risk factor in adult AML, its prognostic impact on childhood AML remains to be determined. RESULTS: We studied the prevalence, cytogenetic and mutational features, and outcome impact of CK in a cohort of 284 Chinese children with de novo AML. Thirty-four (12.0%) children met the criteria for CK-AML with atypical CK being more frequent than typical CK featured with -5/5q-, -7/7q-, and/or 17p aberration. Mutational prevalence was low and co-occurrence mutants were uncommon. Children with CK-AML showed shorter overall survival (OS) (5-year OS: 26.7 ± 10.6% vs. 37.5 ± 8.6%, p = 0.053) and event-free survival (EFS) (5-year EFS: 26.7 ± 10.6% vs. 38.8 ± 8.6%, p = 0.039) compared with those with intermediate-risk genetics. Typical CK tended to correlate with a decreased OS than atypical CK (5-year OS: 0 vs. 33 ± 12.7%.; p = 0.084), and CK with ≥ 5 cytogenetic aberrations was associated with an inferior survival compared with CK with ≤ 4 aberrations (5-year OS: 13.6 ± 11.7% vs. 50.0 ± 18.6%; p = 0.040; 5-year EFS: 13.6 ± 11.7% vs. 50.0 ± 18.6%; p = 0.048). CONCLUSION: Our results demonstrate CK as an adverse risk factor for reduced survival in childhood AML. Our findings shed light on the cytogenetic and mutational profile of childhood CK-AML and would inform refinement of risk stratification in childhood AML to improve outcomes.

Screening of nutritional and genetic anemias using elastic light scattering.

Anemia affects more than » of the world's population, mostly concentrated in low-resource areas, and carries serious health risks. Yet current screening methods are inadequate due to their inability to separate iron deficiency anemia (IDA) from genetic anemias such as thalassemia trait (TT), thus preventing targeted supplementation of oral iron. Here we present an accurate approach to diagnose anemia and anemia type using measures of pediatric red cell morphology determined through machine learning applied to optical light scattering measurements. A partial least squares model shows that our system can accurately extract mean cell volume, red cell size heterogeneity, and mean cell hemoglobin concentration with high accuracy. These clinical parameters (or the raw data itself) can be submitted to machine learning algorithms such as quadratic discriminants or support vector machines to classify a patient into healthy, IDA, or TT. A clinical trial conducted on 268 Chinese children, of which 49 had IDA and 24 had TT, shows >98% sensitivity and specificity for diagnosing anemia, with 81% sensitivity and 86% specificity for discriminating IDA and TT. The majority of the misdiagnoses are IDA patients with particularly severe anemia, possibly requiring hospital care. Therefore, in a screening paradigm where anyone testing positive for TT is sent to the hospital for gold-standard diagnosis and care, we maximize patient benefit while minimizing use of scarce resources.

Large aperture liquid crystal lens array using a composited alignment layer.

A liquid crystal (LC) lens array with high light control power and a large aperture using a composited alignment layer is proposed. In our design, the alignment layer is not only used for getting a uniform arrangement of LC molecule, but also for getting a lens-like refractive index distribution in the LC layer when a voltage is applied. Through simple technology processes, a tunable focal length LC lens array with a millimeter scale diameter can be achieved. Furthermore, the maximum phase difference of the proposed LC lens array can achieve 105.38π. So, the proposed LC lens array has a high light control power.

Correlation of TET2 SNP rs2454206 with improved survival in children with acute myeloid leukemia featuring intermediate-risk cytogenetics.

Single nucleotide polymorphisms (SNPs) may influence the disease course and outcome of hematologic neoplasms. SNP rs2454206 is common in the TET2 gene, which plays a role in epigenetic regulation of myelopoiesis. Few investigations examined the role of TET2 SNP rs2454206 in acute myeloid leukemia (AML) and none of those studies was performed in Chinese populations. Here, we report the prevalence and clinical relevance of TET2 SNP rs2454206 in 254 Chinese patients with childhood AML. Our data demonstrate that TET2 SNP rs2454206AG/GG is associated with improved overall survival and event-free survival in AML patients with intermediate-risk cytogenetics features. The prognostic impact of TET2 SNP rs2454206AG/GG was independent of other common AML risk factors, such as age, white blood cell count, and FLT3-ITD. No difference in TET2 expression levels in AML with TET2 SNP rs2454206AA and TET2 SNP rs2454206AG/GG was detected, indicating that TET2 SNP rs2454206 status does not affect TET2 expression in pediatric AML.

KIT mutations correlate with adverse survival in children with core-binding factor acute myeloid leukemia.

The prevalence and clinical relevance of KIT mutations in childhood core-binding factor (CBF) acute myeloid leukemia (AML) have not been well characterized. In this study, a total of 212 children with de novo AML were enrolled from a Chinese population and 50 (23.5%) of the patients were deemed CBF-AML. KIT mutations were identified in 30% of the CBF-AML cohort. The KIT mutations were clustered in exon 17 and exon 8, and KIT mutations in exons 8 and 17 were correlated with a shorter overall survival (OS) (5-year OS: 30.0 ± 14.5% vs. 73.0 ± 8.5%, p = .007) and event-free survival (EFS) (5-year EFS: 30.0 ± 14.5% vs. 73.0 ± 8.5%, p = .003). Multivariate analysis revealed KIT mutations as an independent risk factor in CBF-AML. Our results suggest that KIT mutations are a molecular marker for an inferior prognosis in pediatric CBF-AML.

A new red cell index and portable RBC analyzer for screening of iron deficiency and Thalassemia minor in a Chinese population.

Anemia is a widespread public health problem with 1/4 ~1/3 of the world's population being affected. In Southeast Asia, Thalassemia trait (TT) and iron deficiency anemia (IDA) are the two most common anemia types and can have a serious impact on quality of life. IDA patients can be treated with iron supplementation, yet TT patients have diminished capacity to process iron. Therefore, distinguishing between types of anemia is essential for effective diagnosis and treatment. Here, we present two advances towards low-cost screening for anemia. First: a new red-cell-based index, Joint Indicator A, to discriminate between IDA, TT, and healthy children in a Chinese population. We collected retrospective data from 384 Chinese children and used discriminant function analysis to determine the best analytic function to separate healthy and diseased groups, achieving 94% sensitivity and 90% specificity, significantly higher than reported indices. This result is achieved using only three red cell parameters: mean cell volume (MCV), red cell distribution width (RDW) and mean cell hemoglobin concentration (MCHC). Our second advance: the development of a low cost, portable red cell analyzer to measure these parameters. Taken together, these two results may help pave the way for widespread screening for nutritional and genetic anemias.

Prognostic significance of P2RY8-CRLF2 and CRLF2 overexpression may vary across risk subgroups of childhood B-cell acute lymphoblastic leukemia.

The cytokine receptor-like factor 2 (CRLF2) gene plays an important role in early B-cell development. Aberrations in CRLF2 activate the JAK-STAT signaling pathway that contributes to B-cell acute lymphoblastic leukemia (B-ALL). The prognostic significance of CRLF2 overexpression and P2RY8-CRLF2 fusion in various B-ALL risk subgroups has not been well established. Two hundred seventy-one patients with newly diagnosed childhood B-ALL were enrolled from a Chinese population. The prevalence of CRLF2 overexpression, CRLF2-P2RY8 fusion, CRLF2 F232C mutation, and JAK2 and IL7R mutational status were analyzed, and the prognostic impact of CRLF2 overexpression and P2RY8-CRLF2 on B-ALL was evaluated by assessing their influence on overall survival and event-free survival. CRLF2 overexpression and P2RY8-CRLF2 were found in 19% and 10%, respectively, in the whole cohort. No correlation between CRLF2 overexpression and P2RY8-CRLF2 was observed. CRLF2 F322C and IL7R mutations were not detected in B-ALL cases overexpressing CRLF2, and no JAK2 mutations were found in the whole cohort either. The results showed that CRLF2 overexpression and P2RY8-CRLF2 were associated with a poor outcome in unselected B-ALL. Moreover, in an intermediate risk B-ALL subgroup P2RY8-CRLF2 was correlated with worse survival, whereas in high- and low-risk subgroups, CRLF2 overexpression predicted a poor outcome. Our findings suggest that P2RY8-CRLF2 is an independent prognostic indicator in intermediate risk B-ALL, while CRLF2 overexpression is correlated with an inferior outcome in high- or low-risk B-ALL. Our study demonstrates that the impact of P2RY8-CRLF2 and CRLF2 overexpression on B-ALL survival may differ across risk subgroups. © 2016 Wiley Periodicals, Inc.

WT1 mutations and single nucleotide polymorphism rs16754 analysis of patients with pediatric acute myeloid leukemia in a Chinese population.

Acute myeloid leukemia (AML) is relatively rare in children. Somatic mutations including the single nucleotide polymorphism (SNP) rs16754 in Wilms tumor 1 gene (WT1) and their prognostic relevance in pediatric AML have not been studied in Chinese populations. We analyzed WT1 mutations and rs16754 genotypes in a cohort of 86 patients with de novo pediatric AML in a Chinese population. We detected WT1 mutations in approximately 20% of the patients. Most of the mutations identified were deletions and insertions clustered in exons 7 and 9. No differences were observed with respect to overall survival and relapse-free survival between patients with and without WT1 mutations. The analysis of rs16754 in WT1 exon 7 revealed G as the major allele. Patients with the rs16754(GG) genotype had improved overall survival (p =0.020) and relapse-free survival (p =0.025) compared with those with either rs16754(GA) or rs16754(AA). Moreover, better overall survival (p =0.044) and relapse-free survival (p =0.068) were observed among patients with wild-type CEBPA with rs16754(GG) compared with those carrying rs16754(GA/AA).

[Effect of simvastatin on rat pulmonary hypertension induced by high pulmonary blood flow and injected monocrotaline].

OBJECTIVE: To explore the therapeutic effect of simvastatin on the pulmonary hypertension induced by injected monocrotaline combining with high pulmonary blood flow of rats. METHODS: Forty male SD rats were randomly divided into the placebo group, simvastatin 1 mg/(kg x d) group, simvastatin 2 mg/(kg x d) group and control group. The arterial-venous shunt rats were injected with monocrotaline at day 7 and developing the pulmonary hypertension after shunting operation. Rats were received either placebo or treatment with simvastatin in the respective group. On day 37 after the shunt operation, the mean pulmonary artery pressure (mPAP) and right ventricular hypertrophy Cright ventricle/(left ventricle+septum), RV/(LV+S)) of all rats were measured. The percent vascular wall thickness and muscularization of the pulmonary small arteries were evaluated. The blood samples were collected to compare the total cholesterol levels between groups. RESULTS: The RV/ (LV+S) and mPAP elevated obviously in placebo group compared to the control group (P<0. 05). The muscularization of pulmonary small arteries and pulmonary artery medial hypertrophy increased significantly in placebo group compared with the control group (P<0. 05). The simvastatin treatment made the attenuation of the above-mentioned indexes in this animal model (P<0. 05). There was no significant statistics difference of the total cholesterol levels between any two groups (P > 0. 05). CONCLUSION: By inhibiting the pulmonary vascular remodeling, simvastatin attenuated the development of pulmonary hypertension. Simvastatin may have preventive and therapeutic effects on pulmonary hypertension.

[The change of mast cells and macrophages in the lung of rat with pulmonary hypertension].

OBJECTIVE: To investigate the change of mast cells and macrophages in lung tissue of rats with pulmonary hypertension (PH) and the effect of simvastatin on it. METHODS: Pulmonary hypertension was established as follow, a shunt between abdominal aorta and inferior vena cava was created in rats, 8 days later, the rats were injected with monocrotaline (60 mg/kg). Moreover, a subgroup of rats were given simvastatin 2 mg/ (kg x d). The mean pulmonary artery pressure (mPAP) and right ventricular weight were measured, and the ratio of right ventricle/left ventricle plus septum CRV/(LV+S)) was calculated. The LSAB method was used to stain anti-tryptase and ED1 in the lung tissue of the rats. RESULTS: In comparision with the control group, mPAP and RV/ (LV+S) of PH rats increased significantly (P<0.05). The RV/(LV+S) in simvastatin group was lower than that in PH rats (P<0.05). The amount of mast cells and macrophages in PH rats were more than that in control group (P<0.05). The amount of mast cells in simvastatin intervention group decreased in comparision with the PH group while the macrophages showed no difference in simvastatin group (P>0.05). CONCLUSION: Mast cells and macrophages may be involved in the development of PH and/or the lesion caused by PH accelerate the accumulation and activation of mast cell and macrophages. Simvastatin has a preventive effect on rat PH, it inhibits mast cell proliferation may be one of mechanism.