[Analysis of Relationship between Coagulation Function and Blood Transfusion in Patients with Emergency Rescue].

Objective: Through researching preoperative coagulation function in the case of ABO-identical blood insufficient for emergency rescue transfusion according to recommended programs of special emergency rescue transfusion was carried out, the relationship between volume of blood products and coagulation function was analyzed.

Methods: The surgical cases of blood transfusion more than 1 600 ml during operation were collected in our hospitals from Aug 2015 to Dec 2016（n=218）, these cases were divided into the normal coagulation group（Group A） and abnormal coagulation group（Group B）, and the patients of emergency rescue transfusion O type blood group（Group C）. The basic information of cases, the infused volume of red blood cell（RBC）, virus-inactivated frozen plasma（VIFP）, fresh frozen plasma（FFP）, cryoprecipitate（C）and platelets（P）, prothrombin time（PT）, activated partial thromboplastin time（APTT）, fibrinogen（FIB）and international normalized ratio（INR）were analyzed, the relationship between volume of blood transfusion and coagulation function were also analysed.

[Experimental Study on Neonatal ABO or RhD Compatible Blood Transfusion].

Objective: To investigate the safety and effectiveness of neonatal ABO or Rh(D) by using compatible blood transfusion through retrospective analysis of data from cases received compatible blood transfusion and type matched blood transfusion.

Methods: The clinical data of 26 cases of neonatal compatible blood transfusion in Chinese Nanchang area from January 2014 to October 2016 were collected, and 26 cases of neonatal type-matched blood transfusion were selected according to ratio of 1:1 cases. The efficiency and safety index of 26 patients compatible blood transfusion were compared with that of type-matched blood transfusion.

[Study on RHCE genotyping of Rh blood group in Uygur nationality of Xinjiang in China].

This study was aimed to investigate the characteristics of RHCE genotyping of Xinjiang Uygur nationality population in China. Primers for detecting RHCE genes were designed according to the references, 89 Uygur nationality RhD-negative samples, 233 Han nationality RhD-negative samples and 109 Han nationality RhD-positive samples were detected by sequence-specific primer-polymerase chain reaction (SSP-PCR) for RHCE genotyping. All above-mentioned samples were unrelated.

[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.

[Investigation of the characteristics of Rh blood group of Uygur nationality in Xinjiang].

The study was to investigate the characteristics of Rh blood group of Uygur nationality in Xinjiang. 1 230 blood samples of Uygur nationality were studied by Rh serological typing, modified antiglobulin test, chloroform/trichloroethylene absorption elution test, upstream, downstream and hybrid Rhesus boxes, 10 exons of D gene, RHD(psi) pseudogene. The results showed that the frequency of RHD negative was 5.

[Screening analysis of irregular antibodies from random donor population in Shaoguan area].

The study was purposed to analyze the frequency and distribution of irregular antibodies in Shaoguan area. Screening 15 033 random donor antibodies in Shaoguan area by screening cells, polybrene and antiglobulin tests. The results indicated that the irregular antibodies were found in 42 samples.

[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.

[A method for Rhesus box test].

To study the method for Rhesus box test and its significance, the sequence specific primers of upstream, downstream and hybrid Rhesus boxes were designed according to RhD gene sequence; the upstream, downstream and hybrid Rhesus boxes were determined by PCP-SSP and mismatched PCR. The results showed that this method was confirmed by DNA Standard test. It was shown that in unrelative RhD positive individuals RHD(+)/RHD(-), RHD(+)/RHD(+) genotype accounted for 9.

[Correlation of newborn hemolytic disease with ABO antibodies in sera of pregnant women].

To investigate the relations between morbidity of hemolytic diseases of newborn and ABO antibodies (HDN) in sera of Han and Yao nationality, pregnant women were examined before and after delivery. Antibodies screen, direct antiglobulin test, free antibodies and elution test were all performed. The results indicated that immunologic ABO antibodies of Han people were found in 673 cases out of 1,471 Han pregnant women, and was also found in 28 cases out of 65 Yao pregnant women, and there was no significant difference of incidences between Han and Yao nationality.

[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.

[Study on detection of samples of Rh-weak D and Del].

To study the detection of weak D and Del from samples initially screened RhD(-), RhD phenotype was initially screened by routine serological test, out of which weak D phenotype was detected by indirect antiglobulin test (IAT) and Del phenotype was detected by chloroform-trichloroethylene absorption-elution test. The results showed that 56 samples were RhD(-) confirmed by routine serology test, which were screened out of 26 200 donors, among them 5 samples were typed as weak D by IAT and 9 cases samples were typed as Del by absorption-elution test. In conclusion, the samples which typed as RhD(-) by routine serological test must be identified by IAT and chloroform-trchloroethylene absorption test is order to detect weak D and Del phenotype.

[Dynamic monitoring of graft-versus-host disease after allogeneic peripheral blood stem cell transplantation by soluble HLA-I antigen detection].

Objective: To assess the value of dynamic monitoring of soluble human lymphocytic antigen-I (sHLA-I) in the prediction of graft-versus-host disease (GVHD) after allogeneic peripheral blood stem cell transplantation (PBSCT).

Method: Sandwich enzyme-linked immunosorbent assay (ELISA) was used to quantitatively detect serum sHLA-I. The serum samples for testing were added into W6 32 monoclonal antibody-coated microtiter plate and incubated with anti-beta2m HRP followed by color development with the addition of the substrate.

[Association of interleukin-6 induction and the novel gene LX3].

Objective: To analyze the association between the expression of the novel gene LX3 and interleukin-6 (IL-6) induction, and explore a new target gene for action mechanism of IL-6.

Methods: The total RNA was extracted from U937 cells induced by IL-6 at different concentrations and varied lengths of time. Reverse transcriptional (RT)-PCR and Northern blotting were employed to determine the expression of LX3 and IL-6 induction.

[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.

[Observation of antibody screen of patients with autoimmune hemolytic anemia].

To observe of alloantibodies of patients with autoimmune hemolytic anemia (AIHA), the alloantibodies masked by autoiantibody were detected by using chloroquine elution test, and the specificity of autoantibody was identified by ether elution test. The results showed that 19 cases out of 38 patients with AIHA were positive detected by indirect antiglobulin test and in 7 cases of them alloantibodies in sera cases were found (1 case of anti-D, 4 cases of anti-E and 2 cases of anti-CE), in 5 cases of them alloantibody were detected carried blood group specificity (3 cases of anti-E, anti-C and anti-c 1 case respectively). In conclusion, detections of alloantibodies by chloroquine elution test and ether elution test were very important for transfusion safety in therapy of patients with AIHA.

[Study on blood ABO typing in patients with autoimmune hemolytic anemia].

To explore effect of autoantibody on identification of ABO and RhD blood group, the blood samples of 38 patients with autoimmune hemolytic anemia (AIHA) were identified by routine typing and typing after chloroquine elution test as well as PCR. The results showed that out of 38 patients with AIHA, 11 cases (31.6%) of ABO blood group were difficulty typed, indirect antiglobulin test were positive, and contradiction between cells typing and sera typing were observed.

[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.

[Exon polymorphism of human RHD gene].

Objective: To study exon polymorphism of human RHD gene and investigate the genetic mechanism of RhD-negative individuals.

Methods: PCR using sequence-specific primers (PCR-SSP) was performed on 40 RhD-positive, 120 RhD-negative and 2 weak D blood samples.

Results: All 10 exons could be detected in the 40 RhD-positive and 2 weak D samples.

[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.

[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.

[Research of Typing for HLA-A, -B on Cord Blood Lymphocytes].

Serological typing for HLA-A, -B has been used for a long time. Recently with the developing of molecular biology technologies, HLA-A, -B typing is now turning to genotyping methods. In our study, the capacity of PCR-SSP in solving problems in HLA-A, -B typing with serological methes was evaluated.

[Biological analysis of a novel gene related with interleukin-6].

Objective: To understand the action mechanism of interleukin (IL)-6 through investigation of its related genes.

Methods: Using electronic cloning with IL-6-related expressed sequence tags (ESTs), a novel gene at its full length of 924 bp was acquired. Reverse transcriptase-PCR method was subsequently employed to amplify the cDNA of this gene from the total RNA of human U937 cells, which were previously activated with 100 ng/ml IL-6 for 8 h.

[Separation and cryopreservation of cord blood mononuclear cells].

The influencing factors on cord blood storage after collection and mononuclear cell separation as well as cryopreservation were studied. The mononuclear cell are separated from blood after blood collection, then cryopreserved and washed after thawed. Results showed that the cord blood kept at 4 degrees C or room temperature less than 24 hours after blood collection, mononuclear cell separated by hydroxyethylstarch and 2 centrifugations, mononuclear cell cryopreserved with 50% DMSO and autoplasma from cord blood as protectives and washing the cells after thawing.

Detection of soluble HLA class I antigens by ELISA and determination of its reference value in population of Guangdong province.

Objective: To explore a method for quantitative detection of soluble human leukocyte antigen class I(HLA-I) antigens, with the assistance of which we attempt to define the reference value of the antigen in the population of Guangdong Province.

Methods: Sandwich enzyme-linked immunosorbent assay (ELISA) was employed with W6/32 monoclonal antibody serving as the solid phase antibody and beta2 microglobulin antibody as the first antibody. HRP-labeled second antibody and the substrate were added for enzyme-catalyzed coloring.