[Establishment and Application of a Method for Determination of Glycosyltransferase Activity].

Objective: To establish a method for determination of glycosyltransferase and to explore the enzyme A, B glycosyltransferase activity in human serum so as to lay the foundation for the determination of enzyme level and enzyme activity.

Methods: The glycosyltransferase activity kit was used to draw phosphate standard curves in our laboratory. The A and B glycosyltransferase activity were determined by the standard curves.

[Polymorphism of M, N Allele in MN Blood Group of Chinese Population].

Objective: To detect the base sequences of all exons and part of introns in the GYPA gene of the glycophorin GPA and to investigate the polymorphism of M, N alleles in Chinese population.

Methods: A total of 225 blood sample were randomly colleeted from unrelated Chinese volunteers and were detected by serology techniques. The primers were designed by self, the seguencing of GYPA gene related with sample exon 1-7 full length sequences of bases and intron-1-7 partial sequence was performed, the polymorphism of M, N gene mutation in mucleotide sequence was analysed.

[Alanine solution as enzyme reaction buffer used in A to O blood group conversion].

The aim of this study was to investigate the effect of alanine solution as α-N-acetylgalactosaminidase enzyme reaction buffer on the enzymatic activity of A antigen. The binding ability of α-N-acetylgalactosaminidase with RBC in different reaction buffer such as alanine solution, glycine solution, normal saline (0.9% NaCl), PBS, PCS was detected by Western blot.

[Analysis of HLA haplotype frequency and linkage disequilibrium in patients with acute lymphoblastic leukemia from Northern Chinese Han].

Objective: To analyze the difference between the frequencies of HLA-A-B, B-DRB1 and A-B-DRB1 haplotype, as well as their linkage disequilibrium pattern in patients with acute lymphoblastic leukemia(ALL) and healthy controls from Northern Chinese Han.

Methods: The frequencies of HLA-A-B, B-DRB1, A-B-DR haplotypes and linkage disequilibrium were estimated by Expectation Maximization method based on the genotypes of 643 patients with ALL and 2 0359 unrelated healthy donors, and the statistical significance between the two groups were estimated by chi-square test. Linkage disequilibrium was analyzed with population genetic methods.

[Establishment of the method to induce and measure human IL-2 in vitro].

This study was aimed to establish the quantitative analysis of hIL-2 in culture supernatant by multifunctional Luminex 100. The lymphocytes were separated from ACD-anticoagulated peripheral blood by density gradient method. The lymphocytes were stimulated with PHA for 48 hours, and frozen at -20 degrees C The relative fluorescence units of standard preparations and samples were detected by multifunctional Luminex 100, and the sample concentrations were calculated by standard curve.

[Comparison between genotyping and serological phenotyping in RhCE blood group].

Objective: To genotype the RHCE gene of Hans, Xinjiang's Uigurs and Kazakstans in China, and to compare the results of RHCE genotyping with that of RhCc/Ee phenotyping.

Methods: RHCE genes of 98 Hans with RhD positive and 230 Hans, 72 Uigurs and 18 Kazakstans with RhD/RHD negative were genotyped with PCR-sequence specific primer (SSP) technique.

Results: The results of RHE/RHe genotyping from samples with RhD positive and negative were in accord with that of phenotyping.

[Effect of platelet CD42a modification by mPEG-SPA with different molecular masses].

Objective: To observe the effect platelet antigen modification by mPEG-SPA with different molecular masses.

Methods: Platelet CD42a was modified by 5 kD and 20 kD mPEG-SPA, respectively, and the fluorescence intensity of CD42a was detect by flow cytometry and the three-dimensional structure of CD42a simulated to analyze the distribution of lysine in CD42a molecule.

Results: After platelet CD42a modification by 5 kD and 20 kD mPEG-SPA, the fluorescence intensity of CD42a decreased sharply by 85.

[A comparative research on RHD gene structures of Chinese Han and Uigur population].

Objective: To research comparatively on the RHD gene structures in unrelated RhD negative individuals of Chinese Uigur and Han population.

Methods: The upstream, downstream, hybrid box and 10 exons of RHD gene were detected with sequence specific primer-PCR technique.

Results: The results showed the genotypes of RhD negative individuals to have the significant difference between Chinese Uigur and Han population, that 94.

[Comparison of Rhesus boxes in Hans and Uighurs].

Objective: To study the difference and similarity between Hans and Uighurs in regard to Rhesus box and its significance.

Methods: The sequence specific primers of upstream, downstream and hybrid Rhesus boxes were designed on the basis of RHD gene sequence. The upstream, downstream and hybrid Rhesus boxes were determined by polymerase chain reaction-sequence specific primer(PCP-SSP) and mismatched PCR.

[Determination of human RHD gene rhesus box and its significance].

The aim was to determine RHD zygosity, further to investigate genetic structure of RHD gene, and to predict hemolytic disease of newborn (HDN). The upstream box, downstream box, and hybrid box of RHD gene were determined by PCR-SSP with 4 primers under the same conditions. The results showed that only hybrid box could be determined in RHD(-)/RHD(-) homozygosity.

[Method of detection of soluble HLA-I and soluble HLA-I level alteration in storage blood].

Aim of this study was to develop the detection method of soluble human leukocyte antigens I (sHLA-I) and to explore sHLA-I level alteration in storage blood and its significance. sHLA-I level in sera of 60 Guangdong normal individuals and sHLA-I concentration in blood components from 20 donors quantitatively were detected by sandwich ELISA. The results showed that sensitivity of this assay was 2.

[Observation on gene polymorphism of Rh blood group in Chinese Han nationality].

To observe the gene polymorphism of Rh blood group in unrelated random individuals and families for Chinese Han nationality, polymerase chain reaction-sequence specific primer (PCR-SSP) was used to amplify the Rh C/E gene, RhD gene, exons, intron 2 and 10, insert and Rh Box in 160 blood samples of RhD positive unrelated individuals and 71 samples of RhD negative unrelated individuals and 7 samples of families whose probands were RhD-negative. The results showed that RhD genes of RhD-negative individuals with C antigens were polymorphism, three forms were found for D exon including intact, partial deletion and complete deletion exons. Insert fragments and Rh Box were found in most cases of families whose probands were RhD-negative and its inheritance accorded with the Mendel's Law, and it did not affect the expression of RhD gene.

[Camouflage of HLA-I antigen in lymphocyte surface].

The objective of this study was to investigate the method and effect of blocking the specific reaction between lymphocyte HLA-I antigen and its antibody. The lymphocytes were disposed with 12 mmol/L methoxypolyethelene glycol benzotriazol carbonate (mPEG-BTC) in concentration gradient in PBS (pH 7.4) at 22 degrees C.

[ABO blood group typing for infants and its application for clinical transfusion].

To study the correct method for determining ABO blood types in infants and its influencing factors, blood types of 33 infants under 6 months old were determined by routine serological method, micro-column gel typing system and PCR-SSP genotyping method. Of the 33 cases with discrepant results of ABO blood type by different methods, the blood types of 32 cases were discrepant between red cell and serological typings in the routine serological method, and a false coincidence in 1 case was caused by bacterial infection resulting in B-like antigen. Correct blood typing was obtained in 27 cases with a correct rate of 84.

[Methoxypolyethylene glycol-modified HLA-A2 antigen on lymphocytes].

Objective: To modify the HLA-A2 antigen on the lymphocytes with methoxypolyethylene glycol (mPEG) so as to block the specific binding site for antibody.

Method: Different types of mPEG (all with final concentration of 12 mmol/L) were used at different temperatures in PBS with varied pH values for the modification of the HLA-A2 antigen.

Result: The modification of the antigen was not obviously affected when it was carried out at 4 degrees Celsius or room temperature, but higher temperatures of 30 and 37 degrees Celsius significantly hampered the modification.

Graft survival assessment of MN genotype after allogeneic transplantation of hematopoietic stem cells of umbilical cord blood.

Objective: To establish a simple and practical method to assess the outcome of allogeneic transplantation of hematopoietic stem cells from umbilical cord blood.

Methods: The DNA was extracted from the samples collected from the donor and the receptor separately before and 15 and 300 d after transplantation. MN genotype was determined by PCR with sequence-specific primers (PCR-SSP) with the 2 pairs of specific primers designed and synthesized on the basis of reported MN gene sequence.