Evaluation and Comparison of Thromboelastography and Conventional Coagulation Tests for Blood Coagulation Function in Children with Henoch-Schönlein Purpura / 中国实验血液学杂志

OBJECTIVE@#To evaluate the coagulation function of children with Henoch-Schönlein purpura (HSP) by thromboelastography (TEG) and conventional coagulation tests (CCTs), and to explore the correlation and consistency of the 2 test methods.@*METHODS@#A total of 468 children with HSP were selected from January 2017 to December 2017 in Beijing Children's Hospital, Capital Medical University. The TEG and CCTs data were analyzed to evaluate coagulation function of children with HSP, meanwhile, the coagulation results were analysed the superiority of the 2 test methods was compared by Pearson correlation and Kappa consistency analysis.@*RESULTS@#There were no clinically significant abnormalities practically in HSP children by TEG and CCTs analysis, except for D-dimer level was elevated (t=9.15, P＜0.001). There were no significant changes for coagulation data from, sex comparison of HSP children (P＞0.05 all), but the coagulation reaction time (R), blood clot formation time (K), α-Angle, CI value, fibrinogen, D-dimer and anti-thrombin III in HSP children with different age groups showed difference (P＜0.05 all), and the blood in children aged 0-2 years old tended to be hypercoagulable. The TEG indexes demonstrated no significant difference in coagulation function of children with HSP each other (P＞0.05). However, CCTs data showed that the blood in children with severe kidney involvement were hypercoagulable. Comparision results of the correlation and consistency of TEG and CCTs in detecting coagulation function of HSP children showed that R was weakly correlated with prothrombin time (PT), International Normalized Ratio (INR) and activated partial thromboplastin time (APTT). There were weak correlation between K, α-Angle and Fib (0.1＜|r|＜0.4 all). There was no obvious consistency between them each other (kappa＜0.4 all).@*CONCLUSION@#The overall changes in coagulation function in children with HSP are not obvious, but the hyperfibrinolysis in hypercoagulable state may exists. Furthermore, younger age and severe kidney involvement may cause hypercoagulation in HSP children. The weakly correlation and consistency of TEG and CCTs in detecting coagulation function of HSP children are furtherly confirmation, and the 2 test methods may be irreplaceable.

Frequency of ABO type IgM antibody in 276 infants younger than six months / 中国实验血液学杂志

The aim of study was to explore the frequency of ABO type IgM antibody in infants younger than six months. 309 hospitalized infants younger than six months were selected at first and their EDTA K(3) anticoagulant blood samples were taken. All the infants were divided into five groups: neonates within 1 week as group I; neonates aged 8 to 14 days as group II; neonates aged 15 days to 1 month as group III; infants aged two to 3 months as group IV and infants aged 4 to 6 months as group V. The monocolonal anti-A, anti-B serums, A cells, B cells and O cells were utilized to carried out the blood typing with tube test. The results indicated that from 309 samples tested 33 AB type sample were excluded. Out of the remains of 276 samples, 29 of 46 samples in group I were positive and with the ABO type consistent rate 63% (29/46); 41 of 64 samples in group II were positive and with the ABO type consistent rate 64% (41/64); 47 of 74 samples in group III were positive and with the ABO type consistent rate 63% (47/74); 28 of 45 samples in group IV were positive and with the ABO type consistent rate 62% (28/45); 40 of 47 samples in group V were positive and with the ABO type consistent rate 85%. It is concluded that the ABO type IgM antibody appear in most infants younger than six months and these IgM antibodies may be regarded as the important evidence for ABO typing in infants.

Establishment of method collecting peripheral blood hematopoietic stem/progenitor cells from infants / 中国实验血液学杂志

The aim of this study was to explore a safe method collecting peripheral blood stem/progenitor cell (PBSPC) from the infants of body weight less than 20 kg by using the COBE Spectra Blood Cell Separator through Auto-PBSC procedure. After washing tube by normal saline, one unit of irradiated RBC was infused into the apheresis set. When the collection terminated, only the concentrated RBC in the apheresis set was returned to the infant. The peripheral mononuclear cells (PBMNCs) and CD34+ cells were counted, the cell viability was determined. The results showed that 13 PBSPC collections were carried out successfully from 7 infants of body weight<20 kg. The average count of MNCs was 4.44x10(8)/kg [(3.46-6.45)x10(8)/kg], the CD34+ count was 2.20x10(6)/kg [(1.34-3.79)x10(6)/kg] and the cell viability was 98.45% (97%-100%) respectively. The vital signs of all the infants went smoothly during collection of PBSPCs. In conclusion, with the aid of COBE Spectra blood cell separator and other measures, the collection of PBSPCs from infants of body weight<20 kg is safe and effective, the PBMNCs containing enough PBSPC can be harvested for transplantation.

Slide platelet aggregation test used as a monitor for patient treated with anti-pletelet drugs / 中国实验血液学杂志

The aim was to verify the effectiveness of slide platelet aggregation test (SPAT) to monitor the inhibition effect of anti-platelet drugs. A group of eight healthy volunteers was examined for SPAT value and T(50) (time necessary for reaching 50% of total aggregation) induced by ADP, arachidonic acid (AA) and cationic propyl gallate (c-PG) respectively before and after administration of ASA in dose of 100 mg/day for 3 days. The group of 41 inpatients at the Department of Cardiovascular Disease treated with anti-platelet drugs and the group of 327 healthy blood donors were also examined for SPAT. The SPAT value of healthy volunteer samples stored at room temperature were measured hourly for four hours. The results showed that: (1) no significant difference was detected between the T(50) before and after ASA administration in health volunteer group when ADP was used as inducer, but a significant difference was detected in this group when AA or c-PG was used as inducer. There was significant linear correlation between SPAT value and T(50) induced by c-PG in health volunteer group before and after administration of ASA (r = 0.998, P = 0.000); (2) there was no significant difference between the SPAT value of health volunteer group before administration of ASA and the SPAT value of health blood donors group (P = 0.853), but there was a significant difference between the SPAT values before and after administration of ASA in health volunteer group (P = 0.000). There was significant difference when the SPAT value of the inpatients treated with anti-platelet drugs was compared with that of healthy blood donor group and with that of health volunteer group before and after administration of ASA (P = 0.000). The cut-off value of SPAT in health blood donor group was 44.6 +/- 11.7 seconds, reference value was from 21.1 seconds to 68.0 seconds; (3) there was no significant difference between SPAT values when platelets samples stored at room temperature for 1, 2, 3, 4 hours (P = 0.815). In conclusion, SPAT can rapidly monitor the inhibition effect of anti-platelet drugs and SPAT may have the similar clinic value with T(50) induced by c-PG.

Collection of peripheral blood stem cells from ABO incompatible allogeneic donors by using blood cell separator / 中国实验血液学杂志

To evaluate the yield of the blood cell separator for collection of peripheral blood stem cells (PBSC) from ABO major and (or) minor incompatible allogeneic donors and the feasibility of PBSC component infusion to the recipients without removal of erythrocytes or plasma, the Cobe Spectra (Version 6.1) blood cell separator was utilized to collect PBSC component from 9 allogeneic donors. Of all the donors, 4 were ABO major incompatible, 2 were minor incompatible and the other 3 were both major and minor incompatible to corresponding recipients. In each cycle, different amount of PBSC component was harvested, and the variable volume plasma chased the cells into the bag was adjusted according to the ABO incompatibility. The nucleated cell count, percentage of mononuclear cell, number of CD34(+) cell and percentage of viable cell (trypan blue excluding rate) in the component were detected. At the time of infusion, a series of protective measures to the renal function of recipients were taken. The results showed that apheresis was twice performed on these eight donors to collected enough PBSC for transplantation or cryopreservation, except one apheresis was enough for cell amount needed by transplantation, as the donor's body weight was much heavier than that of the recipient. Altogether 17 apheresises were performed, the mean yield of nucleated cells was 3.77 x 10(10), in which 97% to 99% were mononuclear cells (MNC). The harvested number of CD34(+) cell was 8.62 x 10(6)/kg. All the trypan blue exclusion rate was 100%. In ABO major incompatible or both major and minor incompatible component, there were 8 - 10 ml packed erythrocytes; in ABO minor incompatible component, there were 80 - 120 ml of plasma. These components were infused into the recipients without removal of erythrocytes or plasma and no haemolytic reaction was observed in any recipient, and their hematopoietic functions soon recovered. Results suggest that enough PBSC can be acquired by using blood cell separator Cobe Spectra (Version 6.1), with the modified separation factors, and the collected PBSC component can be safely infused into the ABO incompatible recipients without removal of erythrocytes or plasma.

Effects of preparation and quality control of cryopreserved platelets / 中国实验血液学杂志

The purpose of this study was to establish a set of techniques for cryopreservation of platelets with dimethylsulphoxide (DMSO) to insure high quality of cryopreserved platelet. The methods were as following: (1) DMSO was filtered in stead of being sterilized before infusion into the bag with platelets. (2) The whole blood was centrifuged immediately after blood collection and the attached tube was tied on the top of the bucket. (3) The related centrifugal force was 480 x g, the accelerating and braking grades of the centrifuge for acceleration and deceleration were 9 and 4 respectively. (4) The flow rate of platelet rich plasma (PRP) could not be too high, 80 - 100 ml PRP should be harvested at 1 minute or so. The infusion rate of DMSO into the PRP was 1 ml/min. After the infusion of DMSO, the PRP bag must be put into the -80 degrees C ultra low freezer at once to make the product to be freezed as soon as possible. The cryopreserved platelet should be thawed in the cycling warm water at the temperature of 38 - 40 degrees C. (5) After thawing of platelet, the platelet, red blood cell and white blood cell were counted, and the bacteria culturing, tests for HBsAg, anti-HCV, anti-HIV, TP and ALT were carried out. The results showed that altogether 14 800 units of cryopreserved platelets were prepared including 80 units collected with blood cell separator, of which quality control was accomplished in 300 units. The manually collected platelet mean count >/= 2.4 x 10(10)/unit, while the apheresis platelet count >/= 2.5 x 10(11)/unit. The yield was over 70%. The contaminated red and white blood cells were </= 1 x 10(9) and </= 1 x 10(7)/unit respectively. All the bacteria cultures were negative, while tests for HBsAg, anti-HCV, anti-HIV and TP were negative too. The ALT values were all in normal range. The transfusion of cryopreserved platelets showed obvious effect of haemostasis. In conclusion, the cryopreserved platelets prepared with this method were of high quality and efficaciousness in haemostasis clinically.

Qualitative analysis of batch preparing cryopreserved fresh platelet rich plasma / 中国实验血液学杂志

To evaluate the efficiency and effectiveness of batch preparing cryopreserved fresh platelet-rich plasma (cryo-FPRP) derived from the volunteer donors, platelet count (Plt), mean platelet volume (MPV), platelet distribution width (PDW), plasma pH, plasma lactic acid concentration, and lactic dehydrogenase (LDH) concentration, germiculture, CD62p positive rate, PAC-1 positive rate, and the fluorescence intensity of platelet GPIb-IX-V were detected in ACD whole blood, fresh platelet-rich plasma (FPRP), FPRP with 5% dimethyl sulphoxide DMSO (DMSO-FPRP), and thawed cryopreserved FPRP (cryo-FPRP); the procoagulant activity of FPRP and cryo-FPR was determinated. The results showed that (1) 70 percentage of platelet were separated from the whole blood into FPRP, and the counts of residual erythrocyte and leucocyte were below 1 x 10(9), and below 1 x 10(7) per unit respectively. (2) The plasma pH, lactic acid concentration and PAC-1 positive rate retained a stable level during the preparing, storing and thawing process. (3) Plasma LDH concentration, platelet CD62p positive rate and GPIb-IX-V concentration in platelet surface were enhanced significantly after being frozen and thawing. (4) The plasma clotting time induced by cryo-FPRP were significantly shorter than that induced by FPRP. It is concluded that: (1) The batch platelet preparing process can efficiently obtain platelet from whole blood donated by volunteer, and the process didn't activate the platelet. (2) Cryopreservation can prevent lactic acid accumulation, pH reduce and activation of GPIIb/IIIa. (3) The membrane of partial platelets are affected by freezing and thawing. (4) The density of GPIb-IX-V complexes in platelet surface and its procoagulant activity are enhanced significantly after the FPRP freezing and thawing process.

Determination and comparison of anti-SARS antibody in children and adults / 中国实验血液学杂志

To investigate the positive rate of anti-SARS antibody in children and adults without SARS, 197 paediatric patients under 14 years old from inpatient and outpatient department of our hospital, 156 healthy children pupils from primary school, 453 adult patients over 18 years old from inpatient and outpatient department of our hospital and other 502 healthy adult blood donors were selected. Anti-SARS antibodies were determined by anti-SARS specific antibody detection kit and ELISA method. The results showed that both the positive rates of IgG antibody in paediatric patients and healthy children were about 2% (4/197 and 3/156), while the positive rates in adult patients and healthy adults were about 0.2% (1/453 and 1/502). The difference between the positive rates of children and adults was significant (chi(2) = 11.61, P < 0.001). IgM antibody was negative in all the samples. It is concluded that the anti-SARS IgG antibody positive rate in children was obvious higher than that in adults. This may be the cause why the cases with SARS in children is much less than in adults.

Micro-column gel indirect anti-globulin technique for screening and identification of irregular antibody / 中国实验血液学杂志

The purpose of this study was to design an antibody screening method based on the micro-column gel indirect anti-globulin technique (MGIAT), using pooled cells and plasma, by comparison with the conventional indirect anti-globulin technique (CIAT) combined with a two-stage papain technique, and to explore the feasibility of the use of plasma instead of serum as test material. The samples of blood recipients in our hospital were screened for irregular antibody using pooled test cells. Screening of the antibodies was identified both by MGIAT and CIAT combined papain technique respectively. The results showed that the irregular erythrocyte antibodies were detected in 20 cases from 5,000 recipients screened by MGIAT, using pooled cells, the positive rate was 0.4%. The specificity of 20 cases of irregular antibodies was as follows: 2 cases of anti-D, 8 cases of anti-E, 1 cases of anti-C, 2 cases of anti-c, 2 cases of anti-Mi(a), 2 cases of anti-Jk(a), 1 case of anti-Le(a) and 2 cases of anti-Fy(a). Antibody was detected from 19 cases using CIAT. Anti-Le(a) was detected with adding complement from Le(a-b-) person. Only 13 cases antibody were found by papain technique. It was concluded that irregular antibody screening by MGIAT using pooled cells can take place of the CIAT combining with papain technique in clinical application. Plasma is superior to serum in antibody screening test.

Detection of the anti-SARS-coronavirus specific antibody levels in 156 SARS patients / 中国实验血液学杂志

The objective of this study was to explore the development of IgG and IgM against SARS CoV and characteristics of changes of antibody titers in patients with severe acute respiratory syndrome (SARS) and to search the opportunity for collecting specific anti-serum from convalescent patients with SARS. The anti-SARS-coronavirus specific antibody levels in 156 SARS patients were measured with ELISA. The results showed that the total positive rates of IgG and IgM were 75.6% and 41.7% respectively, and the negative rate of both IgG and IgM was 23.7%. The average titers of IgG and IgM antibody in positive samples were 18.23 +/- 24.72 and 2.18 +/- 1.13, respectively. There was no significant correlation between the titers of IgG/IgM and sex, age, course of diseases and duration of body temperature recovery. It was concluded that not all SARS patients could produce the anti-SARS-coronavirus specific antibody. The titers of the anti-body are diversified even if the antibodies have been emerged in them. In order to obtain effective anti-serum, the titers of antibody must be tested just before collection of convalescent serum, and it ensures the therapeutic effect and provides a measurable index for clinical transfusion.

Comparison of leucocyte-reduced platelet concentrates produced with spectra version 5.1 and version 7.0 blood cell separators / 中国实验血液学杂志

In the present study, the performance of a new blood cell separator (COBE Spectra LRS Turbo Version 7.0) and that of the previous version LRS version 5.1 in the collection efficiency (CE), collection rate and residual white blood cells during platelet collection from donors were compared. 232 units of platelet concentrates (n = 232) were evaluated and 163 units were collected with the Spectra LRS version 5.1 (Group A) and 69 units with the LRS turbo version 7.0 (Group B). Donor's blood cell counts and parameters, platelet yield, collection efficiency and residual leukocytes in platelet concentrates were analysed. Results showed that the platelet yield was higher in group B than that in group A: (2.90 +/- 1.1) x 10(11) versus (2.58 +/- 1.2) x 10(11), P < 0.001; residual WBCs were less than 5 x 10(6) in 99.4% of group A platelet concentrates and in 97.1% of group B platelet concentrates. Collection efficiency was higher in group B than in group A: 51.4 +/- 8.7 versus 43.6 +/- 6.3. A correlation between platelet count before collecting blood and platelet yield was observed in both groups. In conclusion, the Spectra LRS Turbo version 7.0 showed a higher platelet yield than that with LRS version 5.1. Higher platelet counts before collection allow a higher platelet yield.