Comparison of the performance of two automatic cell morphology analyzers for peripheral-blood leukocyte morphology analysis: Mindray MC-100i and Sysmex DI-60.

INTRODUCTION: To compare the morphological classification ability of peripheral-blood leukocytes of the automatic cell morphology analyzers MC-100i and DI-60. METHODS: (1) MC-100i and DI-60 were used to analyze leukocytes in 432 venous blood samples collected from three tertiary hospitals across China. The preclassification results were compared with the results reported by senior morphological experts (postclassification results) to evaluate the accuracy, sensitivity, specificity, and consistency of leukocyte preclassification for both instruments. (2) In 200 of the 432 blood samples, morphological experts conducted manual microscopic examination for various types of leukocytes. The correlation between the MC-100i and DI-60 leukocyte postclassification results and the expert microscopist results were analyzed. RESULTS: (1) MC-100i preclassified leukocytes and nucleated red blood cells (RBCs). Compared with the postclassification results, the total leukocyte preclassification accuracy of MC-100i was 97.16%, while that of DI-60 was 87.24%. The sensitivity of MC-100i to abnormal cells (including blasts, promyelocytes, neutrophilic myelocytes, neutrophilic metamyelocytes, reactive lymphocytes, abnormal promyelocytes, plasma cells, abnormal lymphocytes and nucleated RBCs) was 90.24%, which was significantly higher than the 50.72% sensitivity of DI-60. (2) Comparing the postclassification results with manual microscopy, except for reactive lymphocytes and basophils, the MC-100i and DI-60 results had good correlations with various leukocyte types and nucleated RBCs (r > 0.85), and MC-100i was better than DI-60 in the recognition of basophils. CONCLUSION: Both MC-100i and DI-60 have good detection ability for five normal types of leukocytes in peripheral blood. MC-100i has significantly better detection sensitivity for abnormal cells in peripheral blood than DI-60.

Plant Traits Guide Species Selection in Vegetation Restoration for Soil and Water Conservation.

Great efforts have been made to improve the soil and water conservation capacity by restoring plant communities in different climatic and land-use types. However, how to select suitable species from local species pools that not only adapt to different site environments, but also achieve certain soil and water conservation capacities is a great challenge in vegetation restoration for practitioners and scientists. So far, little attention has been paid to plant functional response and effect traits related to environment resource and ecosystem functions. In this study, together with soil properties and ecohydrological functions, we measured the seven plant functional traits for the most common species in different restoration communities in a subtropical mountain ecosystem. Multivariate optimization analyses were performed to identify the functional effect types and functional response types based on specific plant traits. We found that the community-weighted means of traits differed significantly among the four community types, and the plant functional traits were strongly linked with soil physicochemical properties and ecohydrological functions. Based on three optimal effect traits (specific leaf area, leaf size, and specific root length) and two response traits (specific leaf area and leaf nitrogen concentration), seven functional effect types in relation to the soil and water conservation capacity (interception of canopy and stemflow, maximum water-holding capacity of litter, maximum water-holding capacity of soil, soil surface runoff, and soil erosion) and two plant functional response types to soil physicochemical properties were identified. The redundancy analysis showed that the sum of all canonical eigenvalues only accounted for 21.6% of the variation in functional response types, which suggests that community effects on soil and water conservation cannot explain the overall structure of community responses related to soil resources. The eight overlapping species between the plant functional response types and functional effect types were ultimately selected as the key species for vegetation restoration. Based on the above results, we offer an ecological basis for choosing the appropriate species based on functional traits, which may be very helpful for practitioners involved in ecological restoration and management.

Soil Moisture and Available Phosphorus as the Factors Driving Variation in Functional Characteristics across Different Restoration Communities in a Subtropical Mountain Ecosystem.

Functional characteristics are increasingly used to evaluate the success of different vegetation restoration. Community functional diversity (FD) and the community-weighted mean (CWM), as two main complementary components, are closely linked to site environment and ecosystem functions. However, the patterns and driving factors of functional characteristics are still not clear in different vegetation restoration types. Here, four community restoration types (secondary shrubland, SL; Pinus yunnanensis forest, PF; mixed needle-broad-leaved forest, MF; natural secondary forest, NSF) were selected to investigate species diversity, FD, CWM, and soil physicochemical properties. The relative effects of species diversity and soil abiotic features on variation in functional characteristics were then evaluated. We found that different restoration communities altered most community structures and functional properties in terms of species diversity, FD, and CWM. CWM values and FD in different communities presented different distribution patterns depending on certain traits and parameters. Significant correlations between functional traits were found at the species and community scales, suggesting a potential covariation between these selected traits in communities. The results of redundancy analysis and variation partitioning showed that most of the variation in functional characteristics, especially CWM, was explained by soil moisture and available phosphorus, indicating that habitat filters regulate the functional characteristics of plant communities mainly by changing the dominant species composition and functional traits of species. Therefore, the selection of restoration species adapted to low soil moisture and available phosphorus and the construction of communities based on selected species as the dominant species can effectively drive community assembly and ecosystem functions in the vegetation restoration process.

A portable smartphone-based hemoglobin point-of-care testing platform for accurate anemia diagnostics.

Anemia affects over 2 billion people worldwide, with the heaviest burden borne by women and children. At present, anemia is diagnosed by measuring hemoglobin (Hb) levels, which must be done in hospitals or commercial laboratories by skilled operators. In this work, we report a portable, affordable ($3), easy-to-operate (1 min) and accurate smartphone-based Hb analyzer (SHbA) that uses a drop of finger-pricked blood for anemia point-of-care test (POCT) applications. POCT of Hb was achieved using a smartphone ambient light sensor (ALS) to accurately measure the absorbance of colorimetric Hb biochemical analysis reagents in a microcuvette, as well as an Android-based application for results analysis. SHbA validation results agreed well with those reported by a hematology analyzer, and the SHbA has an anemia diagnosis sensitivity of 95.4% and specificity of 96.3% for venous blood (n = 360) and a sensitivity of 96.39% and specificity of 95.58% for fingertip blood (n = 475). In addition, SHbA exhibits excellent performance in the diagnosis and treatment guidance of anemia high-risk populations, including tumor chemotherapy patients (n = 424), pregnant women (n = 214) and thalassemia patients (n = 208). Importantly, volunteer self-testing results (n = 20) indicate that SHbA can be used for home-based anemia diagnosis and monitoring. SHbA has the advantages of high sensitivity and specificity while being cheap and easy to operate, making it widely applicable for the diagnosis and treatment of anemia, especially for high-risk patients in areas with poor medical resources.

Screening for malignant tumor cells in serous effusions with an automatic hematology analyzer using a novel diagnostic algorithm.

Background: Due to the high false-positive rate of the high-fluorescence body fluid (HF-BF) cell parameter of the hematology analyzer in BF mode, a novel algorithm based on the Mindray BC-6800 Plus hematology analyzer (BC-6800Plus), with higher diagnostic accuracy compared to that of the traditional HF-BF algorithm, was used to screen for malignant tumor cells in clinical BF samples. In this study, the body fluid mode of BC-6800Plus was applied to investigate the ability of its available parameters and characteristic regional particles in tumor cells screening. Methods: A total of 220 BF samples (including pleural effusion and ascites) were randomly classified into a training cohort (154 samples) and a validation cohort (66 samples), and detected on the BC-6800Plus in BF mode. Based on the scatter plot analysis of the instrument, a novel gating algorithm, malignant cell algorithm-body fluid (MA-BF), was designed to detect the aggregated cells expressing highest fluorescence (FL) signals and side-scatter (SS) signals than other cells. BF collection and analyses were performed in compliance with the CLSI H56-A guideline. tumor cell-positive samples were defined as greater than or equal to confirIIIb (Papanicolaou class system) by the pathological examination. The diagnostic accuracy of HF-BF and MA-BF were determined by the receiver operating characteristic (ROC) curve analysis. Results: When the cutoff values of the absolute count (HF-BF#) and relative count (HF-BF%) were set as 0.022×109/L and 3.0%, respectively, the area under curve (AUC), sensitivity, and specificity were 0.76, 0.85 and 0.55 for HF-BF#, and were 0.70, 0.85, and 0.49 for HF-BF%, respectively. The new parameters, the absolute tumor cell count (MA-BF#) and relative count (MA-BF%), were established in the training cohort using the novel algorithm. We confirmed the cutoff values of MA-HF# and MA-HF% in BF were set as 0.006×109/L and 0.2% in the training cohort, respectively. In the validation cohort, the AUC, sensitivity, and specificity were 0.89, 0.93, and 0.78 for MA-BF#, and were 0.89, 0.87 and 0.75 for MA-BF%, respectively. Conclusions: The MA-BF parameters of the novel algorithm output had better diagnostic accuracy for BF tumor cells than the traditional HF-BF parameters.

Immune microenvironment-related gene mapping predicts immunochemotherapy response and prognosis in diffuse large B-cell lymphoma.

Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin's lymphoma (NHL). The R-CHOP immunochemotherapy regimen is the first-line treatment option for DLBCL patients and has greatly improved the prognosis of DLBCL, making it a curable disease. However, drug resistance or relapse is the main challenge for current DLBCL treatment. Studies have shown that the tumor microenvironment plays an important role in the onset, development, and responsiveness to drugs in DLBCL. Here, we used the CIBERSORT algorithm to resolve the composition of the immune microenvironment of 471 DLBCL patients from the GEO database. We found that activated memory CD4+ T cells and γδ T cells were significantly associated with immunochemotherapy response. Weighted gene co-expression networks (WGCNA) were constructed using differentially expressed genes from immunochemotherapy responders and non-responders. The module most associated with these two types of T cells was defined as hub module. Enrichment analysis of the hub module showed that baseline immune status was significantly stronger in responders than in non-responders. A protein-protein interaction (PPI) network was constructed for hub module to identify hub genes. After survival analysis, five prognosis-related genes (CD3G, CD3D, GNB4, FCHO2, GPR183) were identified and all these genes were significantly negatively associated with PD1. Using our own patient cohort, we validated the efficacy of CD3G and CD3D in predicting immunochemotherapy response. Our study showed that CD3G, CD3D, GNB4, FCHO2, and GPR183 are involved in the regulation of the immune microenvironment of DLBCL. They can be used as biomarkers for predicting immunochemotherapy response and potential therapeutic targets in DLBCL.

Human Amniotic Mesenchymal Stem Cells Inhibit aGVHD by Regulating Balance of Treg and T Effector Cells.

BACKGROUND: Acute graft versus host disease (aGVHD) remains a leading cause of transplant-related mortality following allogeneic haematopoietic cell transplantation (allo-HCT). Human amniotic mesenchymal stem cells (hAMSCs) are a novel mesenchymal stem cells (MSCs), which have stronger proliferation and immunomodulatory ability compared with bone marrow mesenchymal stem cells (BM-MSCs). Besides, as the amniotic membrane is often treated as medical waste after delivery, hAMSCs can be obtained conveniently and noninvasively. The aim of this study was to explore the therapeutic efficacy and underlying mechanisms of hAMSCs transplantation for the humanized aGVHD mouse model. METHODS: We established a humanized aGVHD mouse model by transplanting human peripheral blood mononuclear cells (PBMCs) into NOD-PrkdcscidIL2rγnull (NPG) mice, human amniotic membrane collected from discarded placenta of healthy pregnant women after delivery and hAMSCs were extracted from amniotic membrane and expanded in vitro. Mice were divided into untreated group (Control), aGVHD group (aGVHD), and hAMSCs treatment group (aGVHD+hAMSCs), the hAMSCs labeled with GFP were administered to aGVHD mice to explore the homing ability of hAMSCs. T effector and regulatory T cells (Tregs) levels and cytokines of each group in target organs were detected by flow cytometry and cytometric bead array (CBA), respectively. RESULTS: We successfully established a humanized aGVHD mouse model using NPG mice. The hAMSCs have the ability to inhibit aGVHD in this mouse model through reduced villous blunting and lymphocyte infiltration of the gut while reducing inflammatory edema, tissue destruction and lymphocyte infiltration into the parenchyma of the liver and lung. hAMSCs suppressed CD3+CD4+ T and CD3+CD8+ T cell expression and increased the proportion of Tregs, and besides, hAMSCs can reduce the levels of IL-17A, INF-γ, and TNF in aGVHD target organs. CONCLUSION: The NPG murine environment was capable of activating human T cells to produce aGVHD pathology to mimic aGVHD as in humans. The hAMSCs controlled aGVHD by decreasing inflammatory cytokine secretion within target organs by modulating the balance of Tregs and T effector cells in humanized mice.

Protective effect of Lycium barbarum polysaccharide on di-(2-ethylhexyl) phthalate-induced toxicity in rat liver.

Di-(2-ethylhexyl)-phthalate (DEHP) is the most commonly used plasticizer and it has been a ubiquitous environmental contaminant which affects health. The purpose of this study was to investigate the protective effect of the Lycium barbarum polysaccharide (LBP) at dosages of 100, 200, and 300 mg/kg bw on DEHP-induced (3000 mg/kg) toxicity in rat liver through a 28-day animal experiment. The results showed that LBP attenuated oxidative stress slightly by lowering the production of ROS and improving the activity of SOD and GSH-Px in liver and serum of DEHP treatment rats. At the same time, the levels of PXR, CYP450, CYP2E1, CYP3A1, UGT1, and GST were reduced after LBP treatment. Moreover, LBP decreased the mRNA expression of PXR, UGT1, and GST significantly. These findings suggested that LBP might ameliorate DEHP-induced liver injury by down-regulating the expression of PXR in liver, further down-regulating the downstream phase I and II detoxification enzymes, thus reducing the damage caused by DEHP. Therefore, LBP may have the potential to become an auxiliary therapeutic agent as a natural ingredient of health food.

Intervention mechanism of repeated oral GABA administration on anxiety-like behaviors induced by emotional stress in rats.

The purpose of this study was to investigate the effects and mechanism of repeated oral administration of gamma-aminobutyric acid (GABA) on anxiety-like behaviors induced by emotional stress. Male Sprague-Dawley rats were randomly divided into five groups (8 rats each): control, emotional stress model, three emotional stress + GABA-treated groups (0.5, 1, 2 mg/kg). The rats were given empty water bottles after the training of drinking water to induce emotional stress. Each group was treated with saline or different doses of GABA respectively for 21 consecutive days. Then open field and elevated plus maze were used to assess anxiety-like behaviors. Both frontal cortex and plasma NO metabolites nitrate and nitrite (NOx) levels were determined spectrophotometrically. Results showed that oral administration of GABA significantly reversed the stress-induced anxiety-like negative responses dose-dependently. The frontal cortex NOx levels were lower in stressed rats than in control group (P < 0.05), but higher in 2 mg/kg GABA-treated group than stress model group (P < 0.05). On the other hand, NOx levels in plasma showed a gradual decline trend. Collectively, these results suggest that short repeated oral administration of GABA has an anxiolytic-like effect possibly via preventing NO reduction caused by stress and improving availability of NO in the frontal cortex.

Protective effect of mulberry crude extract against nonylphenol-induced thyroid disruption by inhibiting the activity of deiodinase in rats.

Nonylphenol (NP) is an endocrine-disrupting chemical (EDC) that can lead to thyroid disruption. We explored NP-induced toxicity in the rat thyroid and evaluated the mitigating effects of mulberry crude extract (MCE) on NP toxicity. First, we aimed to evaluate NP-induced thyroid disruption by dosing Sprague-Dawley (SD) rats with NP (0, 30, 90, or 270 mg/kg body weight) daily for 28 days. Second, we aimed to determine whether MCE had a detoxifying effect on NP-induced thyroid disruption by dosing SD rats with NP (270 mg/kg body weight) or/and MCE (30, 60, or 120 mg/kg body weight) daily for 28 days. We found that NP significantly inhibited free triiodothyronin (FT3) and free thyroxine (FT4) activity in rat serum (P < 0.05), but MCE intervention significantly increased FT3 and FT4 serum levels (P < 0.05). It is possible that changes in hormonal composition might trigger the TRH-TSH-TH automatic feedback loop. The activity of the three iodothyronine deiodinases increased significantly after NP-dosing (P < 0.05), but only deiodinase3 (D3) was downregulated after MCE treatment (P < 0.05). Therefore, MCE might be an effective NP-detoxification agent against thyroid disruption because it regulates D3 activity.