Differentiation between Thalassemia Trait and Iron Deficiency Anemia Based on Low Hemoglobin Density and Microcytic Anemia Factor.

BACKGROUND: The most common causes of microcytic hypochromic anemia are thalassemia trait (TT) and iron deficiency anemia (IDA). Clinically, the differential diagnosis of TT and IDA is crucial, but it is typically challenging. Thus, in order to differentiate between TT and IDA, we seek to develop a new discriminative index on an automatic hematology analyzer utilizing the two new RBC characteristics of low hemoglobin density (LHD) and microcytic anemia factor (MAF). METHODS: We recruited a total of 323 subjects, including 115 healthy controls, 83 TT, and 125 IDA. An automated hematology analyzer (DxH800, Beckman Coulter) was used to determine peripheral blood parameters; LHD and MAF were calculated using the parameters of MCHC, Hb, and MCV. The receiver operating characteristic (ROC) curve was used to determine the cutoff values and evaluate the diagnostic value for TT and IDA. RESULTS: LHD was significantly lower in TT than IDA, whereas MAF was higher. To distinguish between TT and IDA, a new formula based on LHD and MAF was developed, with a cutoff value of 0.5, AUC of 0.9706 (95% CI: 0.9503 - 0.9909), and specificity, sensitivity, positive predictive value, and negative predictive values were 92.91%, 91.36%, 89.16%, and 94.40%, respectively. The new formula has proven advantages over conventional indices, such as RDW-SD, MCV, MCH, etc. Conclusions: The RBC parameters LHD and MAF detected by hematology analyzer could be useful for screening for TT and IDA. Our new formula outperforms other discriminant formulas in the literature with high sensitivity and specificity, is simple, rapid, and can aid in early detection and management.

A Resistance-Based Microfluidic Chip for Deterministic Single Cell Trapping Followed by Immunofluorescence Staining.

Microchips are fundamental tools for single-cell analysis. Although various microfluidic methods have been developed for single-cell trapping and analysis, most microchips cannot trap single cells deterministically for further analysis. In this paper, we describe a novel resistance-based microfluidic chip to implement deterministic single-cell trapping followed by immunofluorescence staining based on the least flow resistance principle. The design of a large circular structure before the constriction and the serpentine structure of the main channel made the flow resistance of the main channel higher than that of the trapping channel. Since cells preferred to follow paths with lower flow resistance, this design directed cells into the capture sites and improved single-cell trapping efficiency. We optimized the geometric parameters using numerical simulations. Experiments using A549 and K562 cell lines demonstrated the capability of our chip with (82.7 ± 2.4)% and (84 ± 3.3)% single-cell trapping efficiency, respectively. In addition, cells were immobilized at capture sites by applying the pulling forces at the outlet, which reduced the cell movement and loss and facilitated tracking of the cell in real time during the multistep immunofluorescence staining procedure. Due to the simple operation, high-efficiency single-cell trapping and lower cell loss, the proposed chip is expected to be a potential analytical platform for single tumor cell heterogeneity studies and clinical diagnosis.

The diagnostic model for early detection of gestational diabetes mellitus and gestational diabetic nephropathy.

BACKGROUND: Gestational diabetes mellitus (GDM) and gestational diabetic nephropathy (GDN) have become an increasingly serious problem worldwide, which can cause a large number of adverse pregnancy consequences for mothers and infants. However, the diagnosis of GDM and GDN remains a challenge due to the lack of optimal biomarkers, and the examination has high requirements for patient compliance. We aimed to establish a simple early diagnostic model for GDM and GDN. METHODS: We recruited 50 healthy pregnant (HP), 99 GDM patients, 99 GDN patients at Daping Hospital. Renal function indicators and blood cell indicators were collected for all patients. RESULTS: Compared with HP, GDM, and GDN patients exhibited significantly higher urea/creatinine ratio and NEU. The diagnostic model1 based on the combination of urea/creatinine ratio and NEU was built using logistic regression. Based on receiver operating characteristic curve analysis, the area under the curve (AUC) of the diagnostic model was 0.77 (0.7, 0.84) in distinguishing GDM from HP, and the AUC of the diagnostic model was 0.94 (0.9, 0.97) in distinguishing GDN from HP. Meanwhile, the diagnostic model2 based on the combination of ß2-mG, PLT, and NEU in GDM and GDN patients was built using logistic regression, and the area under the ROC curve (AUC ROC) was 0.79 (0.73, 0.85), which was larger than the individual biomarker AUC. CONCLUSION: Our study demonstrated that the diagnostic model established by the combination of renal function indicators and blood cell indicators could facilitate the differential diagnosis of GDM and GDN patients.

Predictive Value of the Peripheral Blood Parameters for Preeclampsia.

BACKGROUND: Excessive systemic inflammation plays a vital role in pathophysiology of preeclampsia (PE). The aim is to clarify the predictive value of the peripheral blood parameters including white blood cell (WBC), neutrophils, lymphocyte, monocyte, platelet count, mean platelet volume (MPV), plateletcrit, platelet distribution width (PDW), platelet-large cell ratio (PLCR), and the ratio value for PE. METHODS: This retrospective study enrolled 170 PE patients, 123 healthy control pregnant women, and 122 non-pregnant women. When pregnant women were admitted to the hospital for delivery, peripheral complete blood cell count was detected by an automatic blood cell analyzer. Clinical signs and demographic characteristics were recorded. The receiver operating characteristic (ROC) curve was used to determine the cutoff value and analyze the predictive significances for PE. Furthermore, the risk factors of PE were tested by univariate and stratified analyses. RESULTS: This study showed that WBC, neutrophil count, neutrophil percentage, NLR, NMR, and PLR# were significantly increased in PE patients as compared with pregnant control patients (p < 0.001), whereas lymphocyte percentage, monocyte percentage, and PNR were decreased. In addition, there was no significant difference in the rest of the peripheral blood parameters between women with and without PE. The ROC curve result revealed that WBC and neutrophil count had a higher AUC value than the rest of peripheral blood variables. WBC and neutrophil count are positively correlated MAP. Moreover, the WBC and neutrophil count were indicated as independent risk factors for the development of PE. CONCLUSIONS: This study clarifies that peripheral blood parameters of WBC and neutrophil count have good applied value with high sensitivity and specificity in predicting the development of PE and are also independent risk factors for the development of PE.

Enhanced Separation Efficiency and Purity of Circulating Tumor Cells Based on the Combined Effects of Double Sheath Fluids and Inertial Focusing.

Circulating tumor cells (CTCs) play a crucial role in solid tumor metastasis, but obtaining high purity and viability CTCs is a challenging task due to their rarity. Although various works using spiral microchannels to isolate CTCs have been reported, the sorting purity of CTCs has not been significantly improved. Herein, we developed a novel double spiral microchannel for efficient separation and enrichment of intact and high-purity CTCs based on the combined effects of two-stage inertial focusing and particle deflection. Particle deflection relies on the second sheath to produce a deflection of the focused sample flow segment at the end of the first-stage microchannel, allowing larger particles to remain focused and entered the second-stage microchannel while smaller particles moved into the first waste channel. The deflection of the focused sample flow segment was visualized. Testing by a binary mixture of 10.4 and 16.5 µm fluorescent microspheres, it showed 16.5 µm with separation efficiency of 98% and purity of 90% under the second sheath flow rate of 700 µl min-1. In biological experiments, the average purity of spiked CTCs was 74% at a high throughput of 1.5 × 108 cells min-1, and the recovery was more than 91%. Compared to the control group, the viability of separated cells was 99%. Finally, we validated the performance of the double spiral microchannel using clinical cancer blood samples. CTCs with a concentration of 2-28 counts ml-1 were separated from all 12 patients' peripheral blood. Thus, our device could be a robust and label-free liquid biopsy platform in inertial microfluidics for successful application in clinical trials.

Evaluation of the Diagnostic Value of Peripheral Blood Parameters for Neonatal Pneumonia.

BACKGROUND: To evaluate the diagnostic value of peripheral blood parameters including white blood cell (WBC), neutrophil, lymphocyte, monocyte, platelet, mean platelet volume (MPV), platelet distribution width (PDW), mean corpuscular volume (MCV), red cell distribution width (RDW), neutrophil-to-lymphocyte ratio (NLR), monocyte-to-lymphocyte (MLR), platelet-to-lymphocyte ratio (PLR) and platelet-to-neutrophil ratio (PNR) for neonatal pneumonia. METHODS: Two hundred and six full-term neonates in our hospital from January 2018 to December 2019 were enrolled, including 73 pneumonic neonates and 133 health controls. Peripheral blood parameters were measured by an automatic blood cell analyzer. While C-reactive protein (CRP) and PCT concentrations were detected by electrochemical luminescence assay. Clinical signs, characteristic population, temperature, and chest radiograph findings were recorded. The receiver operating characteristic (ROC) curve was used to determine the cutoff values and analyze the diagnosis significances for neonatal pneumonia. RESULTS: This study showed that WBC, neutrophil, RDW, NLR, and MLR levels in the pneumonic group were higher than that of the control group, whereas lymphocyte, monocyte, platelet, and PNR levels were lower (p < 0.05). The ROC curve result showed that NLR and PNR owned higher AUC values than the rest of peripheral blood variables. At a cutoff value 2.581, NLR exhibited 63.01% sensitivity, 90.98% specificity, and 0.847 area under ROC curve (AUC). In addition, at a cutoff value 52.77, PNR showed 84.93% sensitivity, 78.95% specificity, and 0.856 AUC. CONCLUSIONS: This study clarifies that peripheral blood parameter of NLR and PNR have good applied value in diagnosis neonatal pneumonia with high sensitivity and specificity.

Effects of Blood Storage on Cell Population Data.

BACKGROUND: Cell population data (CPD) are leukocyte morphologic parameters, measured by automated hematology analyzers with VCS technology. Many studies have demonstrated that these parameters may have clinical utility for diagnosing or screening certain pathological conditions. This study is to investigate the effects of peripheral blood stored at room temperature on the CPD values and provide useful information about storage-induced CPD changes. METHODS: Venous blood samples from healthy donors kept at room temperature (18 - 25°C) for different time intervals were analyzed using a Coulter DxH800 hematology analyzer. The CPD data collected included mean cellular volume, conductivity and multiple angles of light scatters as well as their corresponding standard deviation for neutrophils, lymphocytes, and monocytes. Peripheral blood smears at each time interval were also prepared and examined microscopically. RESULTS: Peripheral blood kept at room temperature over time significantly affects the CPD values for neutrophils, lymphocytes, and monocytes. These CPD changes are correlated with the morphologic alterations observed under light microcopy, but detected much earlier. Some changes imitate clinical pathological conditions. For example, aged neutrophils showed decreased median angle light scatters, suggesting cytoplasmic degranulation, which can be seen in the case of myelodysplastic syndrome. CONCLUSIONS: This study provides valuable information about storage-induced CPD changes that can affect potential clinical application and interpretation of these automated digital morphologic parameters.