[Feasibility Analysis of Establishing Combat Readiness Blood Bank Based on Low Titer Group O Whole Blood and Group A Plasma].

OBJECTIVE: To explore the feasibility of establishing combat readiness blood bank with low titer group O whole blood and group A plasma. METHODS: The Galileo automatic blood analyzer was used to detect the titers of IgM anti-A and anti-B antibodies in the samples of group O blood donors and IgM anti-B titer in the samples of group A blood donors. Group O blood donors with antibody titers below 128 were selected and included in the mobile blood bank for combat readiness, group A plasma with anti-B titer lower than 128 and group O whole blood with antibody titers below 128 were included in the combat readiness entity blood bank. RESULTS: A total of 1 452 group O blood donors were selected, and the anti-A/B antibody titers were detected. Both antibody titers were distributed below 512, and both peak values of sample distribution were at titer 4. The proportion of samples with titers>128 for both antibodies was relatively low. There was a significant positive correlation between the titers of the two antibodies (r =0.383), and the proportion of samples with IgM anti-A titer higher than IgM anti-B titer was relatively high. 1 335(91.94%) group O blood donors with IgM anti-A and anti-B antibody titers <128 could be included in the mobile blood bank. The anti-B titer of group A blood was detected in 512 cases and the results showed that as the antibody titer increased, the proportion of blood donors gradually decreased. 99.8% of group A blood donors had anti-B antibody titer less than 128, and only one case did not meet the inclusion criteria. CONCLUSION: The proportion of group O blood donors whose whole blood meet the low antibody titer standard is high, and almost all plasma of group A blood donors meet the low titer standard, which improves the blood supply rate in emergencies.

[Platelet Transfusion Strategies for MASPAT-Matched Platelet Transfusion Failed Patient with Allogeneic Hematopoietic Stem Cell Transplantation].

OBJECTIVE: To investigate the causes of ineffectiveness of platelet transfusion with monoclonal antibody solid phase platelet antibody test (MASPAT) matching in patients with allogeneic hematopoietic stem cell transplantation and explore the strategies of platelet transfusion. METHODS: A case of donor-specific HLA antibodies (DSA) induced by transfusion which ultimately resulted in transplantation failure and ineffective platelet transfusion with MASPAT matching was selected, and the causes of ineffective platelet transfusion and platelet transfusion strategy were retrospectively analyzed. RESULTS: The 32-year-old female patient was diagnosed as acute myeloid leukemia (high risk) in another hospital with the main symptoms of fever and leukopenia, who should be admitted for hematopoietic stem cell transplantation after remission by chemotherapy. In the course of chemotherapy, DSA was generated due to platelet transfusion, and had HLA gene loci incompatible with the donor of the first transplant, leading to the failure of the first transplant. The patient received platelet transfusion for several times before and after transplantation, and the results showed that the effective rate of MASPAT matched platelet transfusion was only 35.3%. Further analysis showed that the reason for the ineffective platelet transfusion was due to the missed detection of antibodies by MASPAT method. During the second hematopoietic stem cell transplantation, the DSA-negative donor was selected, and the matching platelets but ineffective transfusion during the primary transplantation were avoided. Finally, the patient was successfully transplanted and discharged from hospital. CONCLUSIONS: DSA can cause graft failure or render the graft ineffective. For the platelet transfusion of patients with DSA, the platelet transfusion strategy with matching type only using MASPAT method will miss the detection of antibodies, resulting in invalid platelet transfusion.

[Correlation Analysis of Hemolytic Transfusion Reaction Induced by Low Titer Antibody].

OBJECTIVE: To establish the diagnostic process of low titer blood group antibody in the occurrence of adverse reactions of hemolytic transfusion. METHODS: Acid elusion test, enzyme method and PEG method were used for antibody identification. Combined with the patient's clinical symptoms and relevant inspection indexes, the irregular antibodies leading to hemolysis were detected. RESULTS: The patient's irregular antibody screening was positive, and it was determined that there was anti-Lea antibody in the serum. After the transfusion reaction, the low titer anti-E antibody was detected by enhanced test. The patient's Rh typing was Ccee, while the transfused red blood cells were ccEE. The new and old samples of the patient were matched with the transfused red blood cells by PEG method, and the major were incompatible. The evidence of hemolytic transfusion reaction was found. CONCLUSION: Antibodies with low titer in serum are not easy to be detected, which often lead to severe hemolytic transfusion reaction.

[Research Progress of in Vitro Experiment of Allergic Transfusion Reaction for Plasma Transfusion and Its Relative Mechanism---Review].

Allergic transfusion reaction (ATR) caused by plasma transfusion is one of the main adverse transfusion reactions, and severe allergic reactions may even endanger the patient's life. Currently, ATR is mainly prevented and controlled by drug prevention and symptomatic treatment, and there still lack of preventive measures such as in vitro experiments. It has been shown that mast cells and basophils are the main effector cells of allergic reactions, and histamine is one of the main mediators of IgE-mediated allergic reactions. Some experiments can be used to identify patients with allergies or plasma components containing allergens, such as detection of serum-specific IgE, IgA, anti-IgA antibody, tryptase and histamine, mast cell degranulation test, basophil activation test, and so on. The basophil activation test can also be used for functional matching of plasma in vitro. Research of in vitro experiment of ATR is good for directing the precise infusion of plasma, reducing waste of resources, and avoiding the risk of blood transfusion. As a pre-transfusion laboratory test for clinical use, in vitro experiment of functional matching provides a new way to prevent ATR.

[Retrospective Analysis of Serological Characteristics and Distribution of Patients with Mimicking Antibodies].

OBJECTIVE: To understand the characteristics of patients with mimicking specificity autoantibodies through the analysis of the causes of autoantibodies, specificity of antibodies, strategy of blood transfusion, effect of transfusion and distribution of antibodies in China and abroad. METHODS: A total of 23 patients who applied for blood in our hospital from January 2017 to June 2019 were identified as mimicking specificity autoantibodies by antibody identification or absorption-elution test. The causes of mimicking specificity autoantibodies, antibody specificity, blood transfusion strategy and blood transfusion effect were analyzed. The relevant articles on antibodies published in China and abroad were summarized and sorted out, and the distribution of antibodies was analyzed. RESULTS: All the 23 patients with mimicking specificity autoantibodies were Rh blood group system antibodies, of which mimicking anti-Ce autoantibodies were the most common (34.8%), followed by mimicking anti-e autoantibodies (26.1%), mimicking anti-D autoantibodies (21.7%), mimicking anti-C autoantibodies (8.7%) and mimicking anti-E autoantibodies (8.7%). Except for 2 cases with suspected history of blood transfusion, the other 21 cases had a history of blood transfusion / pregnancy. The most common cause of mimicking autoantibodies was drug, followed by infection and autoimmune diseases. The hemoglobin (Hb) of pretransfusion in the blood transfusion group was (48.4±23.9) g/L, which was significantly lower than (86.0±38.9) g/L in the non-transfusion group (P<0.01). Except for 2 cases who could not evaluate the effect of blood transfusion, the effective rate of transfusion was 100%. According to the retrospective statistics of 32 related articles published in China and abroad, the most type of mimicking antibodies were in Rh blood group system, accounting for 79.28%, among which anti-E was the main part of all mimicking autoantibodies, accounting for 21.95%. The following ones were in Kidd system MNSs system, and Kell system. CONCLUSION: Combined with the clinical symptoms and the degree of difficulty of blood matching, the best strategy of blood transfusion should be selected to ensure the safety of blood transfusion.

[Detection of Autoimmune Hemolytic Anemia Induced by Salvianolate and Evaluation of Blood Transfusion Efficacy].

OBJECTIVE: To evaluate the characteristics of autoimmune hemolytic anemia caused by salvianolate by antibody detection and clinical index monitoring. METHODS: Micro-column gel anti-human globulin method was used for irregular antibody screening and antibody identification. Salvianolate, sodium creatine phosphate and levocarnitine were used to sensitize red blood cells that were compatible with the patient's plasma, and the RBCs were used to test drug antibody in patient plasma respectively. The patient's clinical examination of hemolysis index and blood transfusion effect were analyed retrospectively. RESULTS: The patients were positive for irregular antibody screening, and there were antoanti-Ce antibodies in serum. The erythrocytes sensitized with salvianolate in the patient's serum were positive, while those sensitized with sodium creatine phosphate and levocarnitine were negative. CONCLUSION: Salvianolate causes drug-induced autoimmune hemolytic anemia in this patient.

Prevalence and Specificity of Red Blood Cell Alloantibodies in Patients from China During 1994-2013.

OBJECTIVE: To analyze the data about red blood cell alloantibodies in patients from mainland China and to provide evidence for formulating a management guideline. METHODS: The Chinese and English literatures about Chinese patients in mainland China published in periodicals were retrieved by CHKD, CNKI, CMJD and PubMed using the key words as unexpected antibody, irregular antibody, blood group antibody, hemolytic transfusion reaction (HTR), hemolytic disease of the newborn (HDN), hemolytic disease of the fetus and newborn (HDFN). RESULTS: A total of 5582 red blood cell alloantibodies were retrieved from 4800 patients. The average prevalence of alloantibody in 89 retrospective analysis reports was 0.34 %. Among all study patients, the 10 most common antibodies were anti-E (33.9%), anti-D (18.3%), anti-c (10.9%), anti-M (9.9%), anti-C (8.1%), anti-e (4.8%), anti-Le(a) (3.4%), anti-P1 (2.0%), anti-Mur (1.6%), and anti-Jk(a) (1.2%). Out of all 136 patients with HTR, the most frequentl alloantibodies were Rhesus antibodies (71.7%), and other antibodies included anti-Jk(b) (5.9%), anti-Le(a) (5.1%), anti-Jk(a) (3.7%), anti-M (1.5%), and anti-Mur (1.5%). A total of 644 alloantibodies contributing to HDFN come primarily from the Rhesus (93.1%) and MNS (6.0%) blood group systems. CONCLUSION: The postnatal Rh prophylaxis should become a routine procedure in mainland China. The use of blood matched for C, E, c, e, Jk(a) and Jk(b) should be recommended for Chinese patients with a history of multiple transfusions. Patients with MNS alloantibodies should be given sufficient attention, and Mur+ red blood cells should be included in antibody screening panels.

[Effect of 25 Gy (60)Co Irradiation on the Physico-chemical Property and Functions of the Platelets During Storage].

OBJECTIVE: To evaluate the effects of the 25 Gy 6°Co irradiation on the physiological and biochemical properties and the functions of the platelets during storage. METHODS: A total of 15 bags of platelets were apheresis-collected from 15 healthy donors, and each bag of platelets were divided into 2 parts, then the platelets were divided into the control group (without 25 Gy 6°Co irradiation) and the irradiated group (with 25 Gy 6°Co irradiation) groups. The two groups of platelets were kept under the condition of (22 ± 2) °C and shaken. The Platelet count and pH value were detected on the d 1, d 2, d 3, d 4 and d 5. The variables such as R, K values, α angle and maximal amplitude (MA) were measured by thrombelastography on the same days. Hypotonic shock response (HSR), morphological score were devised. RESULTS: There were no statistically significant difference in Plt counts, mean platelet volume (MPV), platelet distribute width (PDW) and pH between the two groups (P > 0.05), and Plt count decreased on the end of storage. There were no marked changes in HSR level and morphological score between the two groups during storage, and there were no significant difference between the two groups (P > 0.05). In the TEG analysis there were no significant difference of the R, K, α angle and MA values between the two groups (P > 0.05). R value showed upward trend increased along with prolongation of preserved time (P < 0.01), no significant changes in α angle (P > 0.05), K value was slightly higher and MA value was lower in the last day of storage than the days 1-4 (P < 0.01), respectively. CONCLUSION: 25 Gy 6°Co gamma-ray irradiation can not damage the physiological, biochemical properties and the functions of the platelets during storage. In order to ensure the best curative effect, it is suggested that no matter the platelets were irradiated or not, the platelets should be used as soon as possible.

[Serological characteristics and transfusion efficacy evaluation in 61 cases of autoimmune hemolytic anemia].

This study was aimed to analyze the serological characteristics, efficacy and safety of incompatible RBC transfusion in patients with autoimmune hemolytic anemia (AIHA). The patients with idiopathic or secondary AIHA were analyzed retrospectively, then the serological characteristics and the incidence of adverse transfusion reactions were investigated, and the efficacy and safety of incompatible RBC transfusion were evaluated according to the different autoantibody type and infused different RBC components. The results showed that out of 61 cases of AIHA, 21 cases were idiopathic, and 40 cases were secondary. 8 cases (13.1%) had IgM cold autoantibody, 50 cases (82.0%) had IgG warm autoantibody, and 3 cases (4.9%) had IgM and IgG autoantibodies simultaneously. There were 18 cases (29.5%) combined with alloantibodies. After the exclusion of alloantibodies interference, 113 incompatible RBC transfusions were performed for 36 patients with AIHA, total efficiency rate, total partial efficiency rate and total inefficiency rate were 56.6%, 15.1% and 28.3%, respectively. Incompatible RBC transfusions were divided into non-washed RBC group and washed RBC group. The efficiency rate, partial efficiency rate and inefficiency rate in non-washed RBC group were 57.6%, 13.0% and 29.4%, respectively. The efficiency rate, partial efficiency rate and inefficiency rate in washed RBC group were 53.6%, 21.4% and 25.0%, respectively. There was no significant difference of transfusion efficacy (P > 0.05) in two groups. Incompatible RBC transfusions were also divided into IgM cold autoantibody group and IgG warm autoantibody group. The efficiency rate, partial efficiency rate and inefficiency rate in IgM cold autoantibody group were 46.2%, 30.8% and 29.4%, respectively. The efficiency rate, partial efficiency rate and inefficiency rate in IgG warm autoantibody group were 56.7%, 13.4% and 29.9%, respectively. There was no significant difference of transfusion efficacy (P > 0.05 ) in two groups. Hemolytic transfusion reaction was not observed in all incompatible RBC transfusions. It is concluded that the same ABO type of non-washed RBC transfusion and O type washed RBC transfusion are all relatively safe for the AIHA patients with severe anemia after the exclusion of alloantibodies interference. There is no significant difference of transfusion efficacy in two groups. The same ABO type of non-washed RBC transfusion is more convenient and efficient than washed RBC transfusion, and excessive use of type O RBCs can also be avoided.

[Inside quality control for whole blood preservation performed at blood transfusion compatibility testing laboratory].

This study was aimed to establish the technique for preparation and storage of internal quality control pro-ducts by using existing blood sample resources of blood transfusion compatibility testing laboratory. 24 healthy blood donors with group A and RhD-positive were randomly selected, and 4 ml venous blood from these donors were collected, respectively. Based on the use of anticoagulant type, whether to add red blood cell preservation solution and the samples stored at room temperature for 1 or 2 hours daily, 24 specimens were randomly divided into 8 groups by using factorial design methodology. All samples in tube with cap were stored at 4 degrees C, and placed at room temperature for 1 or 2 hours daily. ABO, RhD blood group (recorded on the agglutination strength of the forward and reverse typing), IgM anti-B antibody titer, and free hemoglobin concentration in the supernatant for all samples were detected at 0, 7, 14, 21, 28, 35 days of products preservation. The results indicated that the red blood cell damage from the group used anticoagulants ACD-B and added the MAP red blood cell preservation solution and placed at room temperature 1 hour daily (recorded as A2B2C1 group) was kept minimal, and FHb concentration and FHb increments at each time point were the lowest (p < 0.01), the FHb concentration on 35th day was only (24.5 +/- 84.5) mg/L. There was no significant change of A antigen, D antigen and IgM anti-B antibody response activity and stability in A2B2C1 group during storage for 35 days (p > 0.05). In conclusion, blood transfusion compatibility testing laboratory can use A2B2C1 program established by this study to prepare relatively stable modified whole blood internal quality control products in the existing conditions, which can be effectively preserved and meet the requirements of internal quality control for blood transfusion compatibility testing.

[Establishment of genotyping method for fetal ABO group from pregnant maternal peripheral blood].

This study was aimed to establish a genotyping method to detect ABO group gene of fetus from peripheral blood of pregnant women for prenatal diagnosis of hemolytic disease of newborn (HDN) resulting from ABO blood group incompatibility. 4 pairs of primers were designed according to ABO blood group gene DNA and mRNA sequences. 20 plasma DNA samples from healthy donors were extracted and amplified to explore the best conditions for plasma DNA extraction and PCR amplification. The O group plasma DNA was mixed with A group or B group plasmas by the ratios of 1:1, 2:1, 4:1, 8:1, 10:1, 20:1, 40:1, 100:1 to simulate the status of mixed ABO gene from pregnant maternal blood and to establish the mixed blood group ABO genotyping technology. The pregnant maternal blood samples with more than 30 weeks of gestation were selected for detecting the fetal ABO blood group genotype. The blood samples should be taken as possible as after birth for identification of ABO blood group and evaluation of sensitivity and accuracy of fetal ABO blood group genotyping technology through peripheral blood of pregnant women. The results indicated that the minimal amount of template DNA from single blood plasma for accuracy identification was at least about 0.625 ng, the DNA amount extracted from 500 microl of plasma could meet the requirement for PCR amplification. When the proportion of O group plasma DNA in mixed plasma DNA was

[The effect of leukocyte depletion by filtration on the quality of apheresis platelets].

This study was aimed to investigate the effect of leukocyte depletion by filtration on the quality of apheresis platelets. 20 units of donor apheresis platelets were randomly selected and were preserved with agitation at 20 - 24 degrees C for 24 - 96 hours, then were filtered on polyester flatbed filters. The platelet concentration, mean platelet volume (MPV), volume of apheresis platelets, leukocyte count, pH value, lactate dehydrogenase (LDH) concentration, K(+) concentration and CD62p expression level on surface of platelet membrane, were detected before and after filtration, as well as the rate of leukocyte depletion and platelet loss were calculated. The results showed that the leukocyte count after filtration was remarkably lower than that before filtration (p < 0.001), and the rate of leukocyte depletion was 99.97%. Platelet loss was approximately 8%, and obviously lower than that of the national standard (p < 0.001). MPV, pH value, K(+) and LDH concentration were not significantly different before and after filtration. Compared with platelets before filtration, CD62p expression level after filtration slightly decreased (p > 0.05). CD62p expression on surface of platelet membrane in perfusion fluid obtained from filter plate was obviously higher than that before filtration (p < 0.05). MA of platelet after filtration slightly decreased (p > 0.05). It is concluded that leukocyte and partial activated platelets can be removed efficiently by using polyester flatbed filters, and platelet loss is very low. Filtration does not adversely affect coagulation activity of the platelets in vitro. Apheresis platelets after filtration can fulfil quality requirements to prevent infection of cytomegalovirus and HLA alloimmunization.

[Application of thrombelastography in evaluation of platelet function during storage].

This study was aimed to explore changes of platelet function in vitro during storage by thrombelastography (TEG). 12 units plateletpheresis were randomly selected and stored at 20 to 24 degrees C with agitation. Thrombelastography variable parameters R, K values and maximal amplitude (MA) were measured on 1, 2, 3, 4, 5 days of platelet storage. Platelet concentration, mean platelet volume (MPV), hypotonic shock response (HSR), CD62p expression and CD62p reexpression on platelet surface were detected at the same time. Changes of platelet function in virto were systematically evaluated by above-mentioned indexes. The results showed that MPV augmented slightly with prolongation of preserved time (p > 0.05), and CD62p expression on platelet surface increased remarkably (p < 0.01), while CD62p reexpression decreased gradually (p < 0.01). There were no significant differences in HSR level of platelets during storage (p > 0.05). R value increased with prolongation of preserved time (p < 0.01). There were no obvious changes on K value and alpha Angle during storage (p > 0.05). There were no obvious changes in MA from 1 to 4 days, and MA decreased slightly on day 5 (p < 0.05). It is concluded that there was no significant change in MA and HSR which reflects comprehensive coagulation of platelets during storage. Platelets on the end of storage have excellent function of hemostasis; Thrombelastography parameter MA value can be used as a valuable indicator for evaluation of platelet function in vitro during storage.