Analysis on Factors Influencing Posttransfusion Effectiveness in Plasma Transfusion of 4423 Cases-Times / 中国实验血液学杂志

OBJECTIVE@#To investigate the related factors influencing plasma transfusion efficacy so as to improve the plasma transfusion efficiency.@*METHODS@#According to the clinical symptoms and the laboratorial results, the patients were divided into transfusion efficient and inefficient groups. A total of13090.8 units of plasma were transfused to 4423 patients. The clinical symptoms and the hemorrhage related index per- and pro-transfusion, plasma components sorts, storage time, and the dose of plasma (kg/ml) transfusion were analyzed.@*RESULTS@#The largest transfusion volume of plasma were in intensive care unit (ICU) accounted for 30.36%, the largest blood plasma per patient transfusion was in cardiac surgery (3.96 U). The analysis of transfusion efficiency showed that in terms of patient age, there were difference in transfusion efficiency among the patients with different ages (P＜0.001). The effective transfusion rate in the group of age ＜18 was 53%, which was higher than that in group of age 18-60(41%) and group of age ＞60 (30%); in terms of sex, the effective transfusion rate in female group was higher than that in male group (42% vs 37%) (P＜0.001); in terms of transfusion plasma volume/body weight, there were differences in transfusion efficiency (P＞0.05). The multi-factor logistic regression analysis showed that there was no significant correlation among the plasma sorts, storage time of the plasma pre-transfusion and transfusion efficiency(P＞0.05). The analysis of the non-hemolytic fever reaction caused by plasma transfusion revealed that there was no statistical difference between the plasma and the leukocyte-depleted plasma groups (P＞0.05).@*CONCLUSION@#The plasma transfusion effectiveness relates with age and sex, but not relates with the transfusion plasma voume/body weight, plasma sorts, and the duration of storage.

Bromodomain and extraterminal proteins-a novel class of therapeutic targets in epigenetics / 药学学报

Bromodomain and extraterminal (BET) proteins are a class of proteins that can interpret epigenetic codes and play an important role in regulating gene transcription through identifying and binding acetylated histones or non-histones proteins. The BET inhibitors have emerged with good therapeutic effects in preclinical disease models such as cancer and inflammation. Some of them have entered clinical studies, demonstrating that there is considerable prospect for drug development with BET as a potential therapeutic target. This review briefly describes the structures and functions of the BET proteins, the BET inhibitors in various diseases, as well as molecular mechanisms involved.

Analysis of Factors Influencing Posttransfusion Effectiveness in 26045 Cases of Platelet Transfusion / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the factors influencing platelet transfusion results so as to improve the platelet transfusion efficiency.</p><p><b>METHODS</b>According to the clinical symptoms (bleeding condition is stopped or improved)and the corrected count of increment (CCI), the patients were divided into efficient transfusion and inefficient transfusion groups. A total of 20 671 patients' clinical data and main platelet transfusion parameters in 26 045 tranfusions including platelet count of per- and post- transfusion, platelet component sorts, storage time and transfusion number were analysed.</p><p><b>RESULTS</b>The comparison of platelet transfusion efficiency in age and sex between two groups did not showed statistical difference (P > 0.05), the platelet count before transfusion between two groups showed statistical difference (t = -5.59, P < 0.001) after converting to log, a significant linear correlation did not exist between storage time of the platelet and CCI (corrected count of increment), but there was statistical difference in transfusion efficiency of patients with different diseases. The patients with hematologic diseases showed lower efficiency of platelet transfusion. According to the results of Wilcocon test detection, there was difference between different times of transfusion and transfusion efficiency, that is to say, the transfusion frequency was higher, the transfusion efficency was lower. The Fisher test indicated that the transfusion efficiency of single platelet transfusion was lower than that of transfussed platelet with other blood components (P < 0.01).</p><p><b>CONCLUSION</b>Platelet transfusion efficiency associates with many factors, including different diseases, whether being transfused with other blood components, the platelet count before transfusion, transfusion frequency, but the time of storage does not relate to the transfusion efficacy.</p>

Effect of 25 Gy (60)Co Irradiation on the Physico-chemical Property and Functions of the Platelets During Storage / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To evaluate the effects of the 25 Gy ⁶⁰Co irradiation on the physiological and biochemical properties and the functions of the platelets during storage.</p><p><b>METHODS</b>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 ⁶⁰Co irradiation) and the irradiated group (with 25 Gy ⁶⁰Co 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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>25 Gy ⁶⁰Co 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.</p>

Prevalence and Specificity of Red Blood Cell Alloantibodies in Patients from China During 1994-2013 / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To analyze the data about red blood cell alloantibodies in patients from mainland China and to provide evidence for formulating a management guideline.</p><p><b>METHODS</b>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).</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

Influence of "dosage effect" on unexpected antibody identification / 中国实验血液学杂志

<p><b>OBJECTIVE</b>This study was to investigate the influence of "dosage effect" on unexpected antibody identification and explore its condition, scope and regularity.</p><p><b>METHODS</b>A total of 40 blood recipient samples containing definite unexpected antibodies were selected by column agglutination technology, then AB fresh plasma was used to dilute the samples to obtain different concentrate liquid. After selecting panel cells which show positive with corresponding unexpected antibody in the serum, "single dosage" antigens were distinguished from "double dosage" ones, and then the antigen-antibody reactions were observed between "single dosage" panel cells and respective diluted recipient samples (by column agglutination technology). It's believable that the highest concentration which retains a negative result was choose to evaluate the agglutination strength between "double dosage" panel cells and diluted unexpected antibody, and to observe the difference happened at different "dosage" antigens with unexpressed antibody.</p><p><b>RESULTS</b>Among 40 diluted recipient samples detected by column agglutination technology, the "dosage effect" appeared in 31 diluted samples. There were 30 samples in which the unexpected antibody agglutinated "double dosage" antigens ≤ 2+, while "single dosage" antigens negative. It appeared in another 1 diluted sample, in which the unexpected antibody agglutinated "double dosage" antigens 3+. There were 9 diluted samples in which the unexpected antibody agglutinated panel cells showing negative results (strength was between 1+-3+ before dilution).</p><p><b>CONCLUSIONS</b>When the unexpected antibodies in Rh, MNS, Kidd, Duffy agglutinated "double dosage" antigens ≤ 2+ (by column agglutination technology) , "single dosage" antibody reaction maybe weaken, even be negative, and it may cause the "dosage effect" to interfere the unexpected antibody identification. The "dosage effect" appears in Rh, MNS, Kidd, Duffy blood system usually.</p>

Investigation and analysis of blood transfusion in 1 766 hospitalized trauma patients / 中国实验血液学杂志

<p><b>OBJECTIVE</b>The study was to understand the incidence of traumatic coagulopathy and the clinical blood transfusion in hospitalized trauma patients so as to provide a reference for guiding scientific component transfusion in trauma or surgical patients.</p><p><b>METHODS</b>By using a software "clinical transfusion database" developed by our department, 1 766 trauma cases who suffered traumatic injury and required hospital admission between 2001 and 2012 were retrieved, and out of them 1 211 patients were given transfusion, and the transfusion-related indicators of the patients such as coagulation, hemoglobin levels before transfusion, trauma situation, massive blood transfusion and total blood transfusion were retrospectively analyzed. According total volume of blood usage during hospitalization,1 211 cases with transfusion were divided into three groups: low volume transfusion group ( ≤ 5 U, n = 471), moderate volume transfusion group (5-10 U, n = 449) and high volume transfusion group (>10 U, n = 291), then the difference of indicators among the 3 groups was compared, and the risk factors of high volume transfusion were analyzed.</p><p><b>RESULTS</b>There were 33 cases of coagulopathy and 52 cases of massive transfusion in trauma patients with transfusion. The transfusion rate of trauma patients was about 68.6%. There was no association between the total amount of blood transfusion and surgical grade or whether surgery. The most patients were transfused using two components (plasma and red blood cell), the ratio of plasma to RBC transfused in patients with coagulopathy was approximately 1.0. In high volume transfusion group, there were more younger and male patients with more serious injury, their infection and death were significantly higher than that in other two groups (P < 0.01).</p><p><b>CONCLUSION</b>There were approximately 69% of hospitalized trauma patients require transfusion, the patients in high volume transfusion group have two populations such as middle-aged and young men who was vulnerable to severe trauma mainly caused by accident injury or fall injury and older women who was vulnerable to osteoporotic hip fractures mainly caused by fall injuries. The coagulation disorders in the patients with trauma coagulopathy should be corrected by transfusion with high ratios of plasma to RBC. Massive transfusion (OR = 95.22), hemorrhagic shock (OR = 17.2), trauma coagulopathy (OR = 4.52) are risk factors of high volume transfusion > 10 U, and massive transfusion also is a risk factor of trauma coagulopathy (OR = 16.257). The routine dynamic monitoring of coagulation should be performed for trauma or surgical patients to guide the clinical transfusion scientifically.</p>

Assessment of RBC transfusion volume and its effect on postoperative pulmonary complications in on-pump CABG patients / 中国实验血液学杂志

This study was purposed to investigate the effect of the transfused RBC amount on pulmonary complications after on-pump CABG surgery, and to explore the influencing factors on RBC transfusion volume. 292 adult patients receiving on-pump CABG surgery were divided into non-RBC transfusion group (n = 71), 1-4 U RBC transfusion group (n = 144) and >4 U RBC transfusion group (n = 77). Adjusted multivariable regression analysis was performed to examine the correlation between transfused RBC amount and the odds of pulmonary complications, and multivariable linear regression was used to analyze the influencing factors on RBC transfusion volume. The results showed that compared the three groups, there was the significant difference in postoperative pulmonary complications (1.4% vs 14.6% vs 24.7%, P < 0.001). A stronger and graded correlation was found between transfused RBC amount and pulmonary complications in on-pump CABG patients, the adjusted odds were increased to 1.251 (95% CI: 1.120-1.398, P < 0.001), and influencing factors on RBC transfusion volume were as follows: age (B:0.102; 95% CI: 0.046-0.157, P < 0.001), sex (B:1.825; 95% CI: 0.692-2.957, P = 0.002), preoperative Hct (B:-36.044; 95% CI:-47.724--25.163, P < 0.001), CPB time (B: 0.031; 95% CI:0.013-0.050, P = 0.001) and acute myocardiac infarction (B:2.769; 95% CI: 1.295-4.243, P < 0.001). It is concluded that the transfused RBC amount is related with postoperative pulmonary complications, and the influencing factors on RBC transfusion volume include preoperative Hct, age, acute myocardiac infarction, sex and CPB time.

Preparation of tranexamic acid-loaded porous starch and evaluation of its hemostatic ability / 中国实验血液学杂志

This study was aimed to develop a new generation of ideal hemostatic powder which can be safely, effectively and easily used mainly to first aid anterior to hospital by the synergistic effect of physical and chemical hemostatic mechanisms. The tranexamic acid(TA)-loaded porous starch(PS) (TAPS) was prepared by using PS as carrier and TA as loaded drug component. The absorption property of TAPS was evaluated by water absorption; the hemostatic ability of TAPS was evaluated by test in vitro and in vivo, the blood coagulation time of TAPS was detected by using Lee-white method. The experiment was divided into 3 groups: blank control group, Yunnan Baiyao group and TAPS group, each group with 10 blood samples in vitro test; the 27 SD rats were used to test in vivo, and randomly were divided into 3 groups: PS,Yunnan Baiyao and TAPS, each group consisted of 9 rats for establishing the animal model of liver trauma and detecting the complete hemostasis time. The results showed that the water absorption of PS did not be affected by TA when dose of TA loaded in PS was <0.02 g/g PS. There was no statistic difference in blood coagulation time between TAPS and PS groups(P > 0.05). The complete hemostatic time of TAPS for trauma of left lobe liver was 236.67 ± 55.00 seconds, which was shorter than that of Yunnan Baiyao (340.00 ± 73.48 seconds) and PS (396.67 ± 68.37 seconds) (P < 0.05 and P < 0.01, respectively). It is concluded that PS can load TA and play the hemostatic effect through releasing TA; the TA loading <0.02 g/g PS did not affect the water absorption and pro-coagulation properties. The TA can enhance the hemostatic efficacy of PS, the hemostatic property of TAPS is derived from synergism of physical and chemical hemostatic mechanisms.

Preparation and activity analysis of mouse anti human platelet CD36 monoclonal antibody / 中国实验血液学杂志

This study was purposed to prepare eukaryotic expression vector of recombinant human platelet CD36 gene. The total RNA was extracted from human liver tissue and the cDNA encoding human platelet CD36 antigen extracellular region (Gly30-Asn439) was amplified by RT-PCR. The cDNA was cloned into the prokaryotic expression vector pMD18 and the recombinant vector was transformed into E. coli DH5α. The positive recombinant pMD18-CD36 plasmid was screened. After sequencing, this combinant vector was inserted into the transient eukaryotic expression vector pTE2, the pTE2-s-CD36-10 His transient eukaryotic expression vector was constructed. The recombinant CD36 Gly30-Asn439 expressed by HEK-293 cells was purified with Ni(2+) 2NTA chromatography. The results showed that 1.4 kb cDNA was amplified by RT-PCR, sequencing of the cDNA indicated the sequence was exactly the same to that in Genbank NM_001001547.2. The HEK293 cells with the plasmid were transfected, and SDS-PAGE confirmed that the transfect HEK293 cells expressed the human CD36 antigen extracellular protein fragments. Western-blot showed that the monoclonal antibody could recognize the recombinant CD36 with the sensitivity of 8 ng. It is concluded that the CD36 Gly30-Asn439 can be highly expressed by human embryonic kidney cells (HEK293), and the monoclonal antibody with biological activity has been obtained, which provide the basis for further study on platelet transfusion refractoriness.

Effect of trehalose-loading on red blood cell membrane / 中国实验血液学杂志

This study was purposed to evaluate the effect of trehalose-loading on physiological and biochemistry properties of red blood cell (RBC) membrane. The samples were divided into the control group (RBC without trehalose loading) and the test group (RBC with trehalose loading). Osmotic fragility reaction was used to determine the osmotic fragility change of loaded RBC membrane in NaCl solution of different osmotic concentration. Flow cytometry and deformeter were used to assay the integrality and deformability of the RBC, respectively. The results showed that the NaCl solution osmotic concentrations were 160 mOsm and 121.4 mOsm, respectively when the haemolysis rate was 50% of the control group and the test group. Flow cytometry data demonstrated that incubation of RBC in a hypertonic trehalose solution resulted in a fraction of cells with different complexity that attached to little Annexin V-FITC, and that it could be removed by washing and resuspending the RBC in an iso-osmotic (300 mOsm PBS) medium. The deformability of the loaded RBC descend, the statistical difference was significant between control and test groups (P < 0.01). It is concluded that the membrane physiological and biochemistry stability and membrane integrality of RBC in a hyper osmotic pressure can be retained after trehalose loading.

Correlation of hemogram changes during pregnancy of healthy women with postpartum blood transfusion / 中国实验血液学杂志

This study was aimed to explore the correlation of hemogram changes during pregnancy of healthy women with postpartum blood transfusion. The outpatient and inpatient information of expectant lying-in women in our hospitals was collected, the route blood test, lever and kidney function and blood coagulation function tests were performed from the 4th to the 10th month of pregnancy. The pregnant women without underlying diseases and non-elderly pregnant women with single fetus were selected as the subjects of study. They were divided into postpartum blood transfusion group and non-blood transfusion group. The white blood cell (WBC) count, hemoglobin (Hb) level, platelet (Plt) count, plateletocrit (PCT), mean platelet volume (MPV), platelet distribution width (PDW) were compared in 2 groups. The results showed that 68 cases out of 450 expectant lying-in women received blood transfusion, among them 30 cases with complete data of puerperal transfusion were taken as blood transfusion group, the 28 cases of non transfusion puerperal as control group. There was no significant difference of hemogram changes between the two groups. However, there was a slight decline in Plt count and Hb level of late pregnant women. What is more, there was no correlation between Plt count change and the PCT, MPV and PDW. It is concluded that the changes of hemogram during pregnancy has no correlation with postpartum hemorrhage and blood transfusion in healthy pregnant women, the Plt count and Hb level of pregnant women slightly decline. Nevertheless, PCT, MPV and PDW are within the normal range.

Optimization of lyophilization procedures for freeze-drying of human red blood cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the different parameters of the lyophilization procedures that affect the recovery of the rehydrated red blood cells (RBCs).</p><p><b>METHODS</b>Human RBCs loaded in tubes were cooled with 4 different modes and subjected to water bath at 25 degrees celsius;. The morphological changes of the RBCs were observed to assess the degree of vitrification, and the specimens were placed in the freeze-dryer with the temperature set up at 40, -50, -60, -70 and -80 degrees celsius;. The rates of temperature rise of the main and secondary drying in the lyophilization procedures were compared, and the water residue in the specimens was determined.</p><p><b>RESULTS</b>The protectant did not show ice crystal in the course of freezing and thawing. No significant difference was found in the recovery rate of the rehydrated RBCs freeze-dried at the minimum temperature of -70 degrees celsius; and -80 degrees celsius; (P > 0.05). The E procedure resulted in the maximum recovery of the RBCs (83.14% ± 9.55%) and Hb (85.33% ± 11.42%), showing significant differences from the other groups(P < 0.01 or 0.05). The recovery of the RBCs showed a positive correlation to the water residue in the samples.</p><p><b>CONCLUSION</b>Fast cooling in liquid nitrogen and shelf precooling at -70 degrees celsius; with a moderate rate of temperature rise in lyophylization and a start dry temperature close to the shelf equilibrium temperature produce optimal freeze-drying result of human RBCs.</p>

Inside quality control for whole blood preservation performed at blood transfusion compatibility testing laboratory / 中国实验血液学杂志

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.

Preliminary study on rehydrated conditions for lyophilized human red blood cells / 中国实验血液学杂志

The objective of this study was to investigate the effect of different rehydration conditions on recovery of the lyophilized red blood cells (RBC) so as to optimize the RBC rehydration. The different conditions, including different rehydration solution, the rehydration temperature, volume change rate of the lyophilized RBC rehydrated by the vapor firstly, were studied, the recovery rate and change of physiological and biochemical properties of the rehydrated RBC were detected. The results indicated that the solution of 10% (w/v) PVP40 in PBS showed the best effect, and the RBC recovery rate increased with increasing of rehydration temperature, and the optimal temperature of rehydration was at 37 degrees C. Pre-rehydration in condition of vapor could raise the RBC recovery rate, and promote the MCV and RDW to close to index of the fresh RBC, the deformability of the rehydrated RBC was no serious as compared with RBC preserved in conventional condition, but the activity level of ATP, G-6-PD, SOD, 2, 3-DPG of the rehydrated RBC less decreased. It is concluded that the optimal rehydration conditions for lyophilized RBC are pre-rehydration in the 37 degrees C with vapor firstly, PBS + 10% (w/v) PVP40 rehydration solution and rehydration temperature at 37 degrees C, but the protection of RBC membrane needs to be furtherly studied.

Optimization of composition and concentration for lyophlizing protectant of human red blood cells / 中国实验血液学杂志

This study was purposed to investigate the effect of different compositions and concentrations of lyophilizing protectants on recovery of RBCs and hemoglobin (Hb) after rehydration of lyophilized RBCs. The RBC lyophilizing protectants composed of a series concentrations of PVP, trehalose and different osmotic protectants were applied for protecting lyophilizing process of RBCs, the recovery of RBCs and Hb after rehydration of lyophilized RBCs was detected. The results showed that there were significant differences in loss ratio of RBCs between protectants composed of different compositions and concentrations (p<0.05 or p<0.01). The loss ratio of RBCs in protectant containing 30% PVP40, 150 mmol/L trehalose and 2% BSA was minimum (0.02%), the loss ratio of RBCs in protectant containing 6% PVP 360, 100 mmol/L trehalose and 2% BSA was maximum (0.27%). The difference of effect between 150 and 50 mmol/L trehalose was statistically significant (p<0.01). The recovery rates of RBCs and Hb in protectants contained PVP40 of different concentrations were different after rehydration of lyophilized RBCs. The protectant containing 15% PVP40, 150 mmol/L trehalose and 2% BSA showed optimal protective efficacy for lyophilized RBCs, the recovery rates of RBCs and Hb were 61.29+/-4.11% and 62.49+/-5.91% respectively, which were statistically different from other protectants (p<0.01). The protectants containing glycerol displayed best efficiency in lyophilization too, the recovery rates of RBCs and Hb were 65.97+/-4.52% and 67.24+/-5.94%, respectively. It is concluded that the protectants composed of 0.8 mol/L glycerol, 15% PVP40, 150 mmol/L trehalose and 2% BSA (pH 7.3 ) may be used as the protectant lyophilizing human RBCs in future study.

Optimization of trehalose loading in red blood cells before freeze-drying / 中国实验血液学杂志

The key points for better protection of trehalose in freeze-drying red blood cells (RBCs) are to resolve non-osmosis of trehalose to red blood cells and to make cytoplasmic trehalose to reach effective concentration. This study was aimed to investigate the regularity of loading RBCs with trehalose, screen out optimal loading condition and evaluate the effect of trehalose on physico-chemical parameters of RBCs during the period of loading. The cytoplasmic trehalose concentration in red blood cells, free hemoglobin and ATP level were determined at different incubation temperatures (4, 22 and 37 degrees C), different trehaolse concentrations (0, 200, 400, 600, 800 and 1000 mmol/L) and different incubation times (2, 4, 6, 8 and 10 hours), the cytoplasmic trehalose, free hemoglobin (FHb), hemoglobin (Hb) and mean corpuscular volume (MCV) in fresh RBCs and RBCs stored for 72 hours at 4 degrees C were compared, when loading condition was ensured. The results showed that with increase of incubation temperature, time and extracellular trehalose concentration, the loading of trehalose in RBCs also increased. Under the optimal loading condition, cytoplasmic trehalose concentration and free hemoglobin level of fresh RBCs and RBCs stored for 72 hours at 4 degrees C were 65.505 +/- 6.314 mmol/L, 66.2 +/- 5.002 mmol/L and 6.567 +/- 2.568 g/L, 16.168 +/- 3.922 g/L respectively. It is concluded that the most optimal condition of loading trehalose is that fresh RBCs incubate in 800 mmol/L trehalose solution for 8 hours at 37 degrees C. This condition can result in a efficient cytoplasmic trehalose concentration. The study provides an important basis for long-term preservation of RBCs.

Aggregation after rehydration of lyophilized platelets / 中国实验血液学杂志

This study was aimed to investigate the aggregation of rehydrated-lyophilized platelets. The aggregation rate of fresh and rehydrated-lyophilized platelets were measured by using thrombin, ristocetin, ADP and collagen as inductors and APACT2 aggregameter; the effects of intra- and extra-cellular trehalose on maximum aggregation rate of rehydrated-lyophilized platelets were detected by using ADP as an inductor. The results showed that the aggregation rate of fresh platelets was all about 100%, while aggregation rate of rehydrated lyophilized platelets was (70.17 +/- 7.36)%, (15.3 +/- 2.81)%, (68.67 +/- 6.86)%, (64.67 +/- 11.6)% respectively, when the concentration of thrombin, ristocetin, ADP and collagen was 1 U/ml, 1.6 mg/ml, 20 micromol/L and 2 microg/ml. The maximum aggregation rates of rehydrated-lyophilized platelets in intra- and extra-cellular trehalose, extracellular trehalose and blank control groups were (66.0 +/- 4.69)%, (25.3 +/- 2.42)% and (11.5 +/- 1.87)% (P < 0.01), meanwhile there was significant difference of rehydrated-lyophilized platelet aggregation rate between intra- and extra-cellular trehalose and extracellular trehalose groups (P < 0.01). It is concluded that the concentrations of thrombin (1 U/ml), ristocetin (1.6 mg/ml), ADP (20 micromol/L) and collagen (2 microg/ml) are optimal for platelets aggregation tests, the internal and extracellular trehalose significantly enhance the aggregation of rehydrated-lyophilized platelets.