Genotyping for Kidd, Kell, Duffy, Scianna, and RHCE blood group antigens polymorphisms in Jiangsu Chinese Han / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Molecular testing is more precise compared to serology and has been widely used in genotyping blood group antigens. Single nucleotide polymorphisms (SNPs) of blood group antigens can be determined by the polymerase chain reaction with sequence specific priming (PCR-SSP) assay. Commercial high-throughput platforms can be expensive and are not approved in China. The genotype frequencies of Kidd, Kell, Duffy, Scianna, and RhCE blood group antigens in Jiangsu province were unknown. The aim of this study is sought to detect the genotype frequencies of Kidd, Kell, Duffy, Scianna, and RhCE antigens in Jiangsu Chinese Han using molecular methods with laboratory developed tests.</p><p><b>METHODS</b>DNA was extracted from EDTA-anticoagulated blood samples of 146 voluntary blood donors collected randomly within one month. Standard serologic assay for red blood cell antigens were also performed except the Scianna blood group antigens. PCR-SSP was designed to work under one PCR program to identify the following SNPs: JK1/JK2, KEL1/KEL2, FYA/FYB, SC1/SC2, C/c and E/e.</p><p><b>RESULTS</b>Serologic antigen results were identical to the phenotypes that were predicted from genotyping results. The allele frequencies for Jk*01 and Jk*02 were 0.51 and 0.49, respectively; for Fy*A and Fy*B 0.94 and 0.06; for RHCE*C and RHCE*c 0.68 and 0.32; and for RHCE*E and RHCE*e 0.28 and 0.72. Among 146 blood donors, all were KEL*02/KEL*02 and SC*01/SC*01, indicating allele frequencies for KEL*02 and SC*01 close to 1.00.</p><p><b>CONCLUSIONS</b>The use of PCR-SSP working under the same condition for testing multiple antigens at the same time is practical. This approach can be effective and cost-efficient for small-scale laboratories and in developing counties. These molecular tests can be also used for identifying rare blood types.</p>

Effect of phospholipomannan of Candida albicans on the production of interleukin 6 and interleukin 8 in monocytes / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate whether Candida albicans-native phospholipomannan (PLM) induce an inflammation response through Toll-like receptor(TLRé2 in human acute monocytic leukemia cell line (THP-1) cells.</p><p><b>METHODS</b>Human THP-1 monocytes were challenged with PLM in vitro. The mRNA expressions of TLR2, TLR4, proinflammatory cytokine [interleukin(IL)-6], and chemokine (IL-8) were assayed by real time reverse transcription polymerase chain reaction. The secretions of IL-6 and IL-8 were measured by enzyme-linked immunosorbent assay. The expression of TLR2 was analyzed with Western blot.</p><p><b>RESULTS</b>PLM increased the mRNA expressions and secretions of proinflammatory cytokines (IL-6) and chemokines (IL-8) in THP-1 cells (all P=0.0000). PLM up-regulated the mRNA and protein levels of TLR2 (P=0.0000), whereas the mRNA level of TLR4 was not altered. PLM hydrolyzed with β-D-mannoside manno hydrolase failed to induce gene and protein expressions of TLR2, IL-6, and IL-8. Anti-TLRS-neutralizing antibody blocked the PLM-induced secretions of IL-6 and IL-8 in THP-1 cells (P = 0.0003, P = 0.0010).</p><p><b>CONCLUSION</b>Canidada albicans-native PLM may contribute to the inflammatory responses during Candida infection in a TLR2-dependent manner.</p>

A large-scale survey for rare blood group screening among blood donors in Chinese over Nanjing area / 中国实验血液学杂志

The purpose of this study was to investigate the distribution of 10 rare red blood groups in Chinese Nanjing population, so as to provide compatible rare blood to patients and to create a donor data bank. Jk (a-b-) (Kidd) phenotypes were detected by urea, while H-(H), GPA-(MNS), GPC-(Gerbich), i+ (Ii) and Lub-(Lutheran) phenotypes were detected by monoclonal, polyclonal antibodies with U type 96 well microplate technology. The screening of Jsb- and k-(Kell), Fya-(Duffy), Ok-(Ok), s-(MNS) and Dib-(Digeo) phenotypes were performed by polymerase chain reaction. The results showed that 2 Jk (a-b-) out of 40337 donation samples and 3 Fy (a-b+) out of 1782 donation samples were found, while no other rare blood phenotypes (H-, GPA-, GPC-, Lub-, Ok-, s-, Jsb-, k-, Dib- and i+) were detected. It is concluded that the frequencies of Jk (a-b-) and Fya(a-b+) are 0.0049% and 0.168% respectively. No more rare blood phenotype was found in this screening.

Effects of platelet-derived membrane microparticles on angiogenesis in chick chorioallantoic membranes / 中国实验血液学杂志

This study was purposed to investigate the angiogenesis effect of platelet-derived membrane microparticles (PMPs) in chick chorioallantoic membranes (CAM). Thrombin were adopted to activate the platelets and then PMPs were obtained. PMPs were isolated by high speed centrifugation. Flow cytometry (FCM) was adopted to evaluate the efficiency of thrombin to produce PMPs and BCA method was adopted to evaluate the content of PMPs. PMPs were put into CAM and the effects of PMPs on angiogenesis in CAM were observed. The results indicated that after incubation for 72 hours at the concentration of 80 microg/ml PMPs, the vessel nets in a 'spoked-wheel' pattern were shown around mixed fibrous filter membranes, number of vessel ramification was 112.5 +/- 11.31 and ratio of vessel area/CAM area was 6.19 +/- 1.29%, but there were not localized allantoic vessels developing in the control group, the number of vessel ramification and ratio of vessel area/CAM area in control group were 82.6 +/- 8.05 and 1.78 +/- 0.33 respectively, so there was significant difference between PMP and control groups. In above mentioned conditions, the number of vessel ramification and ratio of vessel area/CAM area in VEGF group were 128.4 +/- 10.02 and 7.44 +/- 1.36 respectively, so there was no difference between PMP and VEGF groups. It is concluded that PMPs show promotive effect on the formation of capillaries in chick chorioallantoic membranes.

Influence of raising oxygen content on function of platelet concentrate during preservation / 中国实验血液学杂志

To explore the influence of raising oxygen (dissolved oxygen) content on function of platelet concentrate, the platelet concentrate was prepared by a CS-3000 plus blood cell separator. Experiments were divided into 2 groups: test group and control group. After raising oxygen content in platelet plasma under sterile operation, the platelet samples of two groups were preserved in oscillator with horizontal oscillation at 22 +/- 2 degrees C. The platelet count, platelet aggregation rate, lactic acid content and CD62p expression level of platelet were detected on 0, 1, 2, 3, 4, 5 days of platelet preservation. The results showed that the platelet count and platelet aggregation rate decreased with prolongation of preserved time, while the lactic acid content and CD62p expression level of platelet increased gradually. Compared with control group, there were significant differences in aggregation rate of platelet preserved for 2-3 days, and in CD62p expression level of platelet preserved for 1-3 days, while significant difference was found in lactic acid content of platelet preserved for 1-3 days. It is concluded that raising content of oxygen in platelet plasma can provide more oxygen to compensate oxygen supply deficiency for platelet metabolism and improve the efficiency of platelet oxygenic metabolism and the quality of platelet during preservation.

Cold storage of platelet suspension by adding trehalose / 中国实验血液学杂志

The study was aimed to explore the trehalose method for storing platelets in cold. (51)Cr-labeling platelet was used to detect the platelet survival. The platelet function in vitro was performed by platelet aggregate analyzer. After treatment with 50 mg/ml trehalose at 37 degrees C for 4 hours, the rabbit platelet concentrates (PC, 2.0 x 10(9)/ml) were stored in 4-8 degrees C refrigeration, the platelet function in vitro and survival of chilled platelets transfused into self-rabbits were observed. The results showed that trehalose could protect the chilled rabbit platelets. After PC stored at 20-24 degrees C and 4-8 degrees C for up to 24 hours, the platelet aggregate in vitro in response to 11.2 micromol/L ADP were (75.3 +/- 9.8)% and (80.5 +/- 12.5)%, the survival of PC stored at 20-24 degrees C and 4-8 degrees C for 24, 48, 72 hours after transfused into self-rabbits were (78.1 +/- 7.9)%, (65.4 +/- 6.7)%, (57.5 +/- 7.2)% and (5.1 +/- 2.5)%, (2.8 +/- 2.0)%, (0.9 +/- 0.8)%, respectively. The PC treated with 50 mg/ml trehalose were remained stable for up to 12 days of refrigerated storage in autologous plasma. The platelet aggregate in vitro in response to 11.2 micromol/L ADP at 12 days after stored in refrigeration was (77.8 +/- 9.5)%, the survival on 24, 48, 72 hours of platelet transfused into self-rabbits were (75.7 +/- 11.0)%, (67.0 +/- 8.5)%, (56.8 +/- 8.0)%, respectively. Compared with control group of storing at 20-24 degrees C for 24 h, P > 0.05. In conclusion, trehalose can protect the chilled blood platelets, prolong the circulation of refrigerated rabbit platelets, and not impair chilled rabbit platelet function.

Application of cationic propyl gallate as inducer of thrombocyte aggregation for evaluating the platelet function of platelet donors / 中国实验血液学杂志

The purpose of study was to investigate the feasibility of the application of cationic propyl gallate (C-PG) as inducer of platelet aggregation for evaluating the platelet function of single-donor plateletpheresis and identifying the incidence of defective platelet function among donors. Experiments were as follows: 3 healthy volunteers' platelet aggregation induced by 100-300 micromol/L C-PG was determined by LG-PABER analyzer to observe the effect of C-PG concentration on platelet aggregation; 30 healthy volunteers' platelet aggregation before and 24 hours after administration of 200-400 mg acetylsalicylic acid (ASA) was examined after induction by 200 micromol/L C-PG for determining the cut-off value to discriminate platelet dysfunction donors; the platelet aggregation of 483 platelet donors was detected and the activated plasma clotting time (APCT) of donors who have deficiency in platelet aggregation was examined for investigating the incidence of defective platelet function among donors. The results showed that platelets were activated by C-PG induction in a dose dependent manner, when concentration of C-PG reached 200 micromol/L, the percentage of platelet aggregation was highest. It significantly decreased after 24 hours with ASA than that before the administration (P < 0.001), especially in 180 seconds induced by C-PG. If cut-off point was fixed on the platelet aggregation < 20% in 180 seconds, donors of platelet dysfunction can be selected effectively. 25 of defective platelet aggregation function among 483 donors were detected, and 11 out of 25 platelet dysfunction donors had the deficiency in procoagulant activity with prolonged APCT. It is concluded that C-PG as inducer of platelet aggregation is feasible to screen the platelet function of donors. Five percent of platelet donors has function defect examined by C-PG as inducer of platelet aggregation.

Effect of vitamin C antioxidative protection on human red blood cells / 中国实验血液学杂志

In order to investigate the effect of antioxidants on human blood, vitamin C was selected and added into plastic blood storage bags with CPD, and stored at 25 degrees C. During 6 days of storage, some indexes as ATP, SOD, MDA, K(+) concentration and superoxide radicals were detected and were compared with control group, The results showed that ATP and SOD activity in whole blood with vitamin C during 6 days of storage was higher then that in control group (P < 0.05, P < 0.01), the MDA and plasma K(+) concentrations in stored whole blood with vitamin C during 6 days of storage were lower than that in control group (P < 0.05, P < 0.01), the superoxide radical concentrations in stored whole blood with vitamin C decreased lower than that in control group (30%). The conclusion was made that vitamin C increases activities of ATP and SOD, decreases concentrations of MDA, plasma K(+) and superoxide radicals during blood preservation.

Evaluation of the effects of glycosylation on in vivo survival of cold-storage human platelets by using rabbit model / 中国实验血液学杂志

To study the effects of glycosylation on survival of cold-storage human platelets by using rabbit model. (51)Cr-labeling platelets were used to detect the platelet storage survival. The human platelets (2.0 x 10(12)/L) treated with 5 g/L uridine diphosphate galactose (UDP-Gal) were stored in 4 degrees C refrigeratory up to 10 days. The survival of human platelets in rabbits whose reticuloendothelial system was inhibited by the administration of ethyl palmitate was monitored in blood drawn at various times after the platelet transfusion. The results showed that the survival rate of platelets was significantly increased in cold-storage human platelets by UDP-Gal treatment. The survival rates of platelets at 2 hours after transfusion into rabbits in groups of fresh platelets group, UDP-Gal + cold platelets group and cold platelets group were (68.9 +/- 8.5)%, (65.4 +/- 8.0)% and (5.0 +/- 2.6)%, respectively. Compared with cold platelets group, significant differences were seen among all groups (P < 0.01). UDP-Gal + cold platelets group had no significant differences compared with fresh platelets group (P > 0.05). It is concluded that UDG-Gal can provide the protective effect on cold-storage human platelets and prolong the survival time of refrigerated human platelets in rabbit model.

Establishment of detection method for HCMVpp65 of the blood donors and its application in blood bank / 中国实验血液学杂志

To establish method suitable to assay HCMVpp65 of the blood donors in blood bank and to supply safe blood to the patients, the immunocytochemical techniques were used, (6 - 8) x 10(6)/ml cells were counted, 50 x 10(3) cells were detected by light microscope, The results showed that 10 positive samples in 103 samples were found, positive rate was 9.71%, among 10 positive samples, 2 samples were still positive in the second detecting. In conclusion, this method is simple, quick and effective, suitable to detect HCMVpp65 of the blood donors in the blood bank.