Peripheral blood was collected in tubes containing EDTA. Blood cell counts were obtained with a hematology analyzer (Mindray, China). To count BM cells, the BM cells were processed to generate single-cell suspensions. Total BM cell numbers were determined using a hematology analyzer (Mindray). The number of different cell types in the BM was calculated based on BM cell counting and flow cytometry analysis with appropriate markers.
Hematology analyzer
Manufactured by Mindray
Sourced in China
The Hematology Analyzer is a diagnostic device used to analyze and count various blood cells, including red blood cells, white blood cells, and platelets. The analyzer provides comprehensive hematology data to support clinical decision-making.
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Lab products found in correlation
Prism 8, by GraphPad (1 mentions)
Balb c mice, by Charles River Laboratories (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Sodium ascorbate, by Merck Group (1 mentions)
Glycerophosphate, by Merck Group (1 mentions)
Alpha minimum essential medium, by Merck Group (1 mentions)
Antibiotic antimycotic, by Merck Group (1 mentions)
12 protocols using hematology analyzer
1
Peripheral Blood and Bone Marrow Cell Profiling
2
Platelet Concentrate Isolation from Whole Blood
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The study got approval from the Research Ethics Commission of General Hospital of Southern Theater Command of PLA. Platelet concentrates were collected from the venous blood of health volunteers. EDTA was used to prevent blood clotting. After the whole blood was collected, it would be centrifuged for 300 x g (10 min) at 22°C to obtain the buffy coat layer and plasma. Another centrifuge for 900 x g (15 min) at 22°C made platelets precipitated. According to the platelet count in the whole blood, the pellet was resuspended in the plasma or specific buffer. The platelets were counted by a hematology analyzer (Mindray, Shen Zhen, China).
3
Hematological and Biochemical Evaluation of Rabbit Sera
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Hematological and biochemical parameters of blood samples collected from the rabbits used to produce the sera were analyzed to verify eventual toxicity derived from the immunization. For this, the samples were collected before each immunization step and submitted to different analyses. Red and white blood cell counts, hemoglobin concentration, and red blood cell indices were determined by using a hematology analyzer (Mindray Headquarters, Shenzhen, China) according to [33 ]. Biochemical parameters included aspartate aminotransferase, creatinine, muscular creatine kinase, urea, total plasma protein, and fibrinogen; these parameters were quantified by means of commercial kits from Bioclin (Belo Horizonte, MG, Brazil) applied in BS-200 Chemistry Analyzer (Mindray Headquarters, Shenzhen, China) and according to [34 ]. The statistical analyses were performed by two-way ANOVA with Tukey’s post-test, and the significance was established at p < 0.05 using the software GraphPad Prism 8.0.2.
4
Hematology Analysis of Blood Samples
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Blood was collected into tubes with EDTA, and the numbers of lymphocytes, eosinophils and basophils were counted using a hematology analyzer (Mindray).
5
Comprehensive Hematological Analysis
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Blood samples were collected in EDTA-coated tubes, and hematological parameters were determined using a Mindray hematology analyzer. The following hematological parameters were analyzed: total and differential white blood cell count (WBC), red blood cells number (RBC), red cell distribution width (RCDW), hematocrit (HCT), hemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), platelet count (PLT), and mean platelet volume (MPV).
6
Comprehensive Blood Analysis Protocol
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Blood samples were collected from the sacrificed animals and centrifuged at 3000 rpm for 20 min to find plasma. Plasma samples were stored at − 80 °C were analyzed for total protein, albumin, creatinine, GGT (gamma glutamyl transferase), ALP (alkaline phosphatase), ACP (acid phosphatase), AST (aspartate transaminase), ALT (alanine transaminase) and LDH (lactate dehydrogenase) using kits purchased from BioSystems S.A. Costa Brava, 30. 08030 Barcelona (Spain). Heparin was used as an anticoagulant in plasma samples and non-coagulated blood was tested, for WBCs, Lymphocytes(Lymph), Monocytes (Mon), Granulocytes (Gran), Lymph %, Mon %, Gran%, RBCs, HGB, HCT, MCV, MCH, MCHC, RDW % by Mindray hematology analyzer.
Plasma samples stored at − 80 °C were analyzed for total protein, albumin, urea and LDH (lactate dehydrogenase) using kits purchased from BioSystems S.A. Costa Brava, 30. 08030 Barcelona (Spain). Heparin was used as an anticoagulant in plasma samples and non-coagulated blood was tested, for WBCs, Lymphocytes(Lymph), Monocytes (Mon), Granulocytes (Gran), Lymph %, Mon %, Gran%, RBCs, HGB, HCT, MCV, MCH, MCHC, RDW % by using Mindray hematology analyzer.
Plasma samples stored at − 80 °C were analyzed for total protein, albumin, urea and LDH (lactate dehydrogenase) using kits purchased from BioSystems S.A. Costa Brava, 30. 08030 Barcelona (Spain). Heparin was used as an anticoagulant in plasma samples and non-coagulated blood was tested, for WBCs, Lymphocytes(Lymph), Monocytes (Mon), Granulocytes (Gran), Lymph %, Mon %, Gran%, RBCs, HGB, HCT, MCV, MCH, MCHC, RDW % by using Mindray hematology analyzer.
7
Comprehensive Liver and Kidney Evaluation
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Parameters’ liver function (ALT, AST and albumin) and kidney function (urea and creatinine) were detected in plasma using colorimetric kits as well as complete blood count (CBC) was determined using a hematology analyzer (Mindray, China).
8
Hypoxia-Induced Physiological Responses in Mice
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Twenty-four healthy male BALB/c mice (20.0–20.6 g; Vital River, Beijing, China) with ad libitum access to food and water were randomly divided into four groups: (1) control; (2) normoxia with RSV; (3) hypoxia; (4) hypoxia with RSV. The hypoxia groups were fed in the hypoxic chamber (Aipu, Hangzhou, China), which was set to 25 ℃ for 7 days. Nitrogen gas was fed at a flow rate of 1 L/min to gradually maintain the oxygen content range at 9.5–10.5%. This was done to simulate the environment at 6000 m above sea level, forming an acute severe hypoxia state. For the RSV group, RSV was injected intraperitoneally at a concentration of 100 mg/kg once daily. The weight was recorded every day. Seven days later, the arterial blood gas was detected using a blood gas analyzer (ABL90 FLEX, Radiometer, Brønshøj, Denmark) and a blood cell analysis was performed using a hematology analyzer (Mindray, Shenzhen, China) immediately after arterial blood was drawn from the carotid artery of each mouse. The changes in PO2, SO2, cLac, RBC count, hematocrit (Hct), and hemoglobin (HGB) values were analyzed and the oxygen supply efficiency indicators (P50, SI, theoretical oxygen-release capacity) were observed. Then, the mice were sacrificed and subjected to H&E pathological analysis to detect changes in the liver, brain, and lungs before and after hypoxia.
9
Quantifying Mesenchymal Stem Cell Enrichment
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Ten milliliters of pre- and post- enrichment bone marrow was collected for MSC adherent culture as reported by other researchers16 (link),17 (link). In brief, samples were cultured in six-well plates with osteogenic induction solution [alpha-minimum essential medium (Sigma, Santa Clara, California, USA) suspended in 10% fetal bovine serum (Hyclone, Logan, Utah, USA), 50 mg/mL sodium ascorbate (Sigma), 1% antibiotic/antimycotic (Sigma), 10 mM glycerophosphate (Sigma), and 10-8 M dexamethasone (Sigma)]. The medium was changed every 2 days, and at 10–14 days, the cultures were washed by phosphate-buffered saline (PBS), fixed by 4% paraformaldehyde and then stained with an alkaline phosphatase (ALP) staining kit (Lexiang, Shanghai, China), which as reported before was used for calculating colony-forming units (CFUs). Each CFU that was 2 mm in diameter or larger was manually quantified as one count. The mean count of ALP-positive colonies from six individual wells was used for each sample, and the presence of ALP+CFU represented an MSC colony18 (link). Enrichment efficiency was formulated as (PREALP+CFUs – POSTALP+CFUs)/ PREALP+CFUs×100%. Another 1 mL bone marrow pre- and post-enrichment was sent for routine blood examination by a hematology analyzer (Mindray, China) to evaluate the adhesion of other cells in bone marrow, including leukocytes, erythrocytes, and platelets.
10
Newly Diagnosed Adult ITP Patients
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This was a retrospectively cohort study. Adult newly diagnosed ITP patients who were hospitalized in the hematology units of our hospital from October 2016 to May 2021 were included. The study was approved by Institutional Review Board of our hospital. All patients enrolled in this study signed the informed consents.
The inclusion criteria were shown as follows: (1) patients who were aged 18 years or older; (2) patients who were diagnosed as primary ITP according to the International Working Group (IWG) diagnostic criteria [10 (link)]; (3) patients who had not received any therapy before. Patients with secondary, relapsed, or refractory ITP were excluded.
The data including age, gender, height, weight, treatment protocol, ITP-specific bleeding score, adverse reactions, platelet glycoprotein-specific antibodies expressions, and treatment responses were retrospectively collected. The routine blood count including platelet count was measured by a hematology analyzer (Mindray Biomedical Electronics Co., Ltd., Shenzhen, China).
The inclusion criteria were shown as follows: (1) patients who were aged 18 years or older; (2) patients who were diagnosed as primary ITP according to the International Working Group (IWG) diagnostic criteria [10 (link)]; (3) patients who had not received any therapy before. Patients with secondary, relapsed, or refractory ITP were excluded.
The data including age, gender, height, weight, treatment protocol, ITP-specific bleeding score, adverse reactions, platelet glycoprotein-specific antibodies expressions, and treatment responses were retrospectively collected. The routine blood count including platelet count was measured by a hematology analyzer (Mindray Biomedical Electronics Co., Ltd., Shenzhen, China).
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