Objective To gain purified platelets for further research of the immune function of platelet.Methods Platelets were purified by positive(MS) and negative(LD) magnetic cell sorting(MACS) separation.The percentage of activated platelets was detected by flow cytometry and nucleated cell clearance was evaluated by white cell count.Results The percentage of activated platelets before separation was(2.39?1.10)%,and increased to(2.56?1.08)% and(16.76?4.04)%,respectively,after MS and LD MACS.The clearances of nucleated cells after MACS MS and LD were(98.44?0.24)% and(98.47?0.18)%,respectively.The recovery rate of purified platelet after MS and LD was(76.50?1.49)% and(79.20?2.61)%.Conclusion MACS LD method was more suitable for the platelet purification.

Objective Study on fetomaternal immuno state and RHD type of a pregnant woman of weak D phenotype.MethodsThrough polymerase chain reaction (PCR)、direct genomic DNA sequencing and flow cytometry.ResultsIn both sequence specific promer (SSP) PCR and the sequencing PCR tests, the sample was detected negative in exons 3 6 of the RHD gene, whereas all other exons (exons 1 2,7 10) were tested positive. And the sequence of detected exons (exons 1 2,7 10) are the same with normal RHD in GenBank (accession no. AJ299020 1 and AJ299026 9). Serologically and genetically, the sample can be designated as D category VI type Ⅲ. Through a duce tube PCR method, the RhD zygosity of this individual was typed CD VI e/cde。In flow cytometry, a few fetal erythrocytes were detected in peripheral blood of the mother. However there were no anti D detected in sera and hemolytic disease of the newborn(HDN) observed at all.ConclusionSevere cases of HDN have occurred in D positive babies born to partial D mother with anti D, although HDN don't take place in this case. We may still consider D VI phenotype individuals as D positive donors and D negative receiptions in our transfusion practice and in clinical anti D allo immune prophylaxis and monitoring.

Objective To prepare B-lymphoblastoid cell lines of HLA novel allele B*5610 in a family for further study and identification . Method Isolate mononuclear cells under aseptic conditions from the peripheral blood. After infection with Epstein-Barr virus, the cells were cultured in 20% FBS, 2?g/ml CsA RPMI 1640. Results Immortalized B-lymphoblastoid cell lines of five B *5610 carriers in a family were achieved, and the new genes were inherited stably. Conclusion Our work is important for storing and breeding the precious material of biomedicine because the B *5610 genes in the immortalized B-lymphoblastoid cell lines were inherited stably.

ObjectiveTo analyze the polymorphism of HLA-A,B,and DRB1 alleles and their haplotypes in Chinese Man population.Methods Frequencies of HLA-A,B and DRB1 alleles and haplotypes were estimated by maximum-likelihood estimation method based on the genotypes of 2183 Chinese Man bone marrow donors.ResultsA total of 18 HLA-A alleles,44 HLA-B alleles and 15 HLA-DRB1 alleles were detected in Man population,and the most frequent alleles were A*02,A*11,A*24,A*30,A*33,B*13,B*35,B*46,B*51,B*40(B60),B*40(B61),B*15(B62),DRB1*04,DRB1*07,DRB1*08,DRB1*09,DRB1*11,DRB1*12,DRB1*13,DRB1*14 and DRB1*15.A*30-B*13,A*02-DRB1*15,B*13-DRB1*07 and A*30-B*13-DRB1*07 were the most frequent haplotypes in Man population for A-B,A-DRB1,B-DRB1 and A-B-DRB1 haplotype,respectively.The number of haplotypes with frequency ≥ 0.01 for A-B,A-DRB1 and B-DRB1 haplotype was 31,24 and 27,respectively,and ≥ 0.005 for A-B-DRB1 haplotype was 32.There were 14 in A-B,3 in A-DRB1,14 in B-DRB1 and 38 in A-B-DRB1 haploypes that showed strong linkage disequilibrium with ALD≥0.40.ConclusionsThe distribution of HLA-A,B and DRB1 alleles and haplotypes in Man population was similar to that in Northern Chinese Han population.

Objective To study the inter-allelic recombination event occurring in the HLA-C locus in a family of Chinese Han nationality, and to evaluate the molecular genetic background of the new HLA allele.Methods Peripheral blood samples were collected from a Chinese leukemia woman patient, as well as her healthy parents and two brothers.HLA-A, C, B, DRB1 and DQB1 alleles were typed by high-resolution PCR-sequence-based typing (SBT) method using Atria Genetic AlleleSEQR HLA SBT kits.The Protrans S4 HLA-C single allele-specific sequencing strategy was used to separate the two HLA-C alleles and to determine novelty of the allele.The full length sequences of HLA-C alleles of the patient and her parents were further analyzed using cloning and haplotype sequencing method. The HLA five loci linked haplotypes and the recombination site were analyzed by family study, meanwhile the full length sequences of the five HLA-C alleles were compared with the IMGT/HLA database by the program BLAST.Results The two haplotypes of the father and mother were a:A*0207-C*010201-B*550201-DRB1*090102-DRQ1*030302 and b:A*240201-C*120202-B*5201-DRB1*1502-DRQ1*0601, c:A*300101-C*060201-B*130201-DRB1*0405-DRQ1*0401 and d:A*110101-C*070201-B*4001-DRB1*080302-DRQ1*0601,respectively.The two brothers inherited their parent′s haplotypes a, d and b, c respectively.The two haplotypes of the patient were the maternal c and paternal recombinant a/b haplotype.The recombinant a/b haplotype A*240201-C*new-B*550201-DRB1*090102-DRQ1*030302, A*240201 came from the paternal haplotype b,while B*550201-DRB1*090102-DRQ1*030302 came from the other paternal haplotype a.When comparing the full length sequences of the HLA-C new allele with the father′s allele C*010201 and C*120202, it could deduce that the recombinant a/b haplotype derived from a recombination event occurring between the paternal chromosome 6 during meiosis.The crossover site was between genomic nt273 and nt330 of HLA-C alleles, which created a HLA-C new allele and the fifth haplotype of the family, and inherited it to the patient.The full length sequences of the new allele had been submitted to Genbank, and officially named C*0121 by WHO nomenclature committee.Conclusion This study demonstrates a rare inter-allelic recombination event occurring in the HLA-C locus within a Chinese Han family and illustrates the process of novel allele and haplotype, and provides direct theory for further studying the mechanisms of gene recombination and HLA polymorphism.

Objective To compare efficacy of female sterilization by modified Uchida technique and silver clips and to evaluate the influence on operation procedure and clinical effect with or without surgery training of service providers. Methods A comparative, multicenter clinical trial was performed in 18 county and township-level service centers. Totally 2198 women underwent sterilization from these 18 study center were divided into 1116 women sterilized by modified Uchida technique and 1082 women by silver clips.Those 18 centers were classified into 9 training groups which provide surgical skills of sterilization and other contents and 9 non training groups. Clinical documents of sterilization were recorded. All women were followup at 3, 6 and 12 months after surgery. Results There were no complications during surgery by both sterilization. The failure rate was 2.03% (22/1082) in silver clip method and the mean operative time were ( 12. 4 ± 6. 4 ) minutes in training group and ( 14. 4 ± 8. 1 ) minutes in non training group. In modified Uchida method, the failure rate was 0. 18% (2/1116) and the mean operative time were (16. 2 ± 4. 9)minutes in training group and (19.0 ±8.6) minutes in non training group. The mean operative time between two groups reached statistical difference ( all P ＜ 0. 05 ). Total ended rate in modified Uchida technique were 2. 2/hundred women year in training group and 2. 5/hundred women year in non training group, and the rate of silver slips were 3. 9/hundred women year and 4. 8/hundred women year, which did not show significant difference ( all P ＞ 0. 05 ). There was no significant difference in acceptability and side effects of all women between two methods (P ＞ 0. 05). The training of service providers could influence acceptability of women (P ＜ 0. 05). Conclusions Clinical efficacy was not influenced by those two methods. The operative time and acceptability were improved by training surgeons in silver clips method.

Objective To explore the mechanism of therapeutic effect of Donepezil hydrochloride on Alzheimer's disease(AD) rats.Methods According to weight,36 rats were divided into normal group,model group and Donepezil hydrochloride group.AD rat model was set up by injecting D-galactose into abdominal cavity for seven weeks,learning and memory function of rats was determined by using Morris water maze and Step-down test.The section of rat cerebral cortex and hippocampus were stained with haematoxylin eosin(HE),and the effect of Donepezil hydrochloride was observed by detecting the MDA content and SOD activity in cerebral tissue.Results Compared with model group,latency and distance of Donepezil hydrochloride rats shortened on the fourth day and the fifth day,starting angle of Donepezil hydrochloride rats shortened on the fourth day and the fifth day in the Morris water maze test,error times of Donepezil hydrochloride rats decreased on the first day and the second day in Step-down test;MDA content in cerebral tissue of Donepezil hydrochloride rat was deceased(P

0.05).Conclusion Donepezil hydrochloride do not improve the learning and memory function of normal under age rats.

Objective To assess the clinical characteristics of postoperative myocardial infarction (PMI) in patients undergoing off-pump coronary artery bypass grafting(OPCAB).Methods Two hundred and sixty-six patients undertook OPCAB in the Shandong Provincial Chest Hospital from January 2008 to June 2015 were selected,there were 22 cases clinical diagnosis of PMI as PMI group,44 cases patients whose general information matched MI group were selected as no PMI group.The data of two groups including preoperative records and postoperative symptoms,electrocardiogram (ECG),cTnI and echocardiography change were compared.Results There were no significant differences about preoperative indexes between the two groups(P ＞0.05).Incidences of severe chest pain and new pathological Q-waves and elevated ST segments were significantly higher in PMI group than those in no PMI group (90.9％ (20/22) vs.18.2％ (8/44),27.3％ (6/22) vs.4.5％(2/44),95.4％(21/22) vs.27.3％(12/44)),the differences were significant(P＜0.005).Peak serum levels of cTnI during the first 24 h after operation were significantly higher in patients of PMI group than those in no PMI group,the difference was significant((4.52±2.81) μg/L vs.(0.26±0.22) μg/L,P=0.04).There was no significant difference in the incidence of myocardial segmental motion.Conclusion It is difficult to predict coronary artery bypass grafting after myocardial infarction.It has great value of postoperative ECG,the patient complained in diagnosis of postoperative myocardial infarction.CTn is a very sensitive indicator,but its diagnosis clinical myocardial infarction boundary value still need to be open to question.