Objective To investigate the distribution of A subgroups in the A and AB blood type populations among voluntary blood donors in Zhongshan,study the antigen-antibody characteristics of A subgroups,establish a local A subgroup database,and support the development of precision medicine.Methods ABO subgroup screening was performed using the microplate method.Specimens negative for monoclonal anti-A1 reactivity underwent sequencing of exons 1～7 of the ABO gene to confirm genotypes.Cryopreservation and thawing of glycerolized subgroup red blood cells(RBCs),as well as preservation efficacy of concentrated human-derived antibodies with preservatives,were studied.Results Among 1 212 blood donor specimens,28 subgroup specimens were identified,with a prevalence of 1.54%(15/971)in blood type A and 5.39%(13/241)in blood type AB.Sequencing of 10 specimens revealed 7 ABO genotypes:ABO*A2.01/O.01.02(2 cases),ABO*A1.02/B.01(3 cases),ABO*BA.02/O.01.02,ABO*AW.31.02-05/A2.05,ABO*A2.05/B.01,ABO*A2new/O.01.01,and ABO*A1.02/O.01.01(1 case each).Additionally,one rare allele mutation(c.700C>G)and one novel allele mutation(c.203G>T)(GenBank accession number:PQ152337)were identified.Human anti-A1 antibodies with a titer of 8 were successfully concentrated.Optimal preservation conditions included 0.1%preservative concentration and cryopreserved subgroup RBCs stored at 4℃for 3 days post-thaw.Conclusion The predominant A subgroups in Zhongshan donors are A2 and B(A).A preliminary database for A2 and A2B subgroups is established,along with the discovery of a novel ABO allele mutation.Cryopreservation with glycerol,PEG antibody concentration,and ProClin 300 preservative demonstrate effective applications in preserving ABO blood group antigens and antibodies.

Objective To investigate the distribution of A subgroups in the A and AB blood type populations among voluntary blood donors in Zhongshan,study the antigen-antibody characteristics of A subgroups,establish a local A subgroup database,and support the development of precision medicine.Methods ABO subgroup screening was performed using the microplate method.Specimens negative for monoclonal anti-A1 reactivity underwent sequencing of exons 1～7 of the ABO gene to confirm genotypes.Cryopreservation and thawing of glycerolized subgroup red blood cells(RBCs),as well as preservation efficacy of concentrated human-derived antibodies with preservatives,were studied.Results Among 1 212 blood donor specimens,28 subgroup specimens were identified,with a prevalence of 1.54%(15/971)in blood type A and 5.39%(13/241)in blood type AB.Sequencing of 10 specimens revealed 7 ABO genotypes:ABO*A2.01/O.01.02(2 cases),ABO*A1.02/B.01(3 cases),ABO*BA.02/O.01.02,ABO*AW.31.02-05/A2.05,ABO*A2.05/B.01,ABO*A2new/O.01.01,and ABO*A1.02/O.01.01(1 case each).Additionally,one rare allele mutation(c.700C>G)and one novel allele mutation(c.203G>T)(GenBank accession number:PQ152337)were identified.Human anti-A1 antibodies with a titer of 8 were successfully concentrated.Optimal preservation conditions included 0.1%preservative concentration and cryopreserved subgroup RBCs stored at 4℃for 3 days post-thaw.Conclusion The predominant A subgroups in Zhongshan donors are A2 and B(A).A preliminary database for A2 and A2B subgroups is established,along with the discovery of a novel ABO allele mutation.Cryopreservation with glycerol,PEG antibody concentration,and ProClin 300 preservative demonstrate effective applications in preserving ABO blood group antigens and antibodies.

Objective:To explore the characteristics of T lymphocyte subsets and cytokines in hyperlipidemia-induced acute pancreatitis (HLAP) and its prognostic value.Methods:This study included 184 patients with acute pancreatitis (AP) admitted to the First Affiliated Hospital of Xiamen University from January 2018 to May 2021. Based on disease etiology, there were 92 HLAP cases and 92 non-hyperlipidemia-induced AP (NHLAP) cases. Stratified by disease severity according to 2012 Atlanta classification criteria, the patients were divided into the severe subgroup (SAP) and non-severe subgroup (NSAP). Peripheral venous blood samples were taken from all patients on day 1, 3, and 5 after admission. T lymphocyte subsets were determined by flow cytometry, and cytokines were detected by flow fluorometry. The number of CD4 +% and CD8 +% and the expression of cytokines were compared by Student’s t test or Mann-Whitney U analysis. Logistic regression analyses were performed to identify risk factors for severe AP, and a receiver operating characteristic (ROC) curve was constructed to predict severe AP. Statistical significance was taken as P<0.05. Results:Compared with the NHLAP group, patients in the HLAP group had lower CD4 +%, while higher levels of IL-2 on day 1 ( P<0.05), and had also lower CD4 +%, while higher levels of IL-4, IL-6, and IL-10 on day 3 ( P<0.05). Furthermore, IL-6 and IL-10 levels of the HLAP group were significantly increased compared to the NHLAP group on day 5 ( P<0.05). IL-10 levels in the SAP subgroup were significantly higher than those in the NSAP subgroup on day 1 ( P<0.05). Compared with the NSAP subgroup, the SAP subgroup had elevated levels of IL-2, IL-4, IL-6, IL-10 and IFN-γ on day 3 (all P<0.05), and had lower CD4 +%, while increased levels of IL-6 and IL-10 on day 5 (all P<0.05). Multivariate Logistic regression analysis showed that IL-10 was an immune indicator of independent risk factor for severe AP in the HLAP group on day 1 ( OR=1.139, 95% CI: 1.038-1.251, P<0.05). Finally, ROC analysis showed that the area under the curve of IL-10 to assess HLAP with severe AP was 0.772, and the best cut-off value for predicting severe AP was 5.6 pg/mL, with a sensitivity of 83.3% and a specificity of 68.8%. Conclusions:Changes of CD4 +% and cytokines are different between the HLAP and NHLAP groups. IL-10 can be used as a predictor of early disease severity in patients with HLAP.

To compare the difference in tumor immunity and autoimmunity elicited by adenovirus (Ad) encoding human or murine tyrosinase-related protein 2 (AdhTRP2 or AdmTRP2), and to find the most effective way to induce immunity by AdhTRP2 or AdmTRP2, C57BL/6 mice were immunized with AdhTRP2 or AdmTRP2 intramuscularly at different doses of 10(5), 10(6), 10(7) and 10(8) separately (10 mice for each dose). Two weeks after the immunization, in vivo CTL assay and intracellular staining (ICS) of IFN-gamma were carried out to analyze the dose-effect relationship. Tumor growth and vitiligo (as an sign of autoimmunity) were observed until 3 months after challenge with 10(5) B16F10 tumor cells. The results showed that Ad encoding AdmTrp2 induced weak tumor immune response. Similar immunization with AdhTrp-2 elicited stronger protective immunity. CTL activity and IFN-gamma-produced CD8+T cells were directly proportional to dose of AdhTrp2 or AdmTrp2. Moreover, AdhTrp2 group showed tumor rejection in 100% of challenged mice till the end of 3rd month while 60% of mice immunized with AdmTrp2 were protected against tumor. In the whole process of this experiment, no vitiligo was observed in mice immunized either with AdhTrp2 or AdmTrp2. It is concluded that anti-melanoma responses induced by genetic vaccination expressing xenoantigens breaks immune tolerance effectively and is able to elicit strong antigen-specific cytotoxic T cell response without vitiligo.
Adenoviridae/metabolism ; Antineoplastic Agents/*pharmacology ; Cell Line, Tumor ; Cytokines/metabolism ; Immune System ; Immune Tolerance ; Interferon-gamma/metabolism ; Intramolecular Oxidoreductases/*biosynthesis ; Intramolecular Oxidoreductases/*genetics ; Mice, Inbred C57BL ; T-Lymphocytes, Cytotoxic/*metabolism ; Vitiligo/*metabolism

To compare the difference in tumor immunity and autoimmunity elicited by adenovirus (Ad) encoding human or murine tyrosinase-related protein 2 (AdhTRP2 or AdmTRP2), and to find the most effective way to induce immunity by AdhTRP2 or AdmTRP2, C57BL/6 mice were im-munized with AdhTRP2 or AdmTRP2 intramuscularly at different doses of 105, 106, 107 and 108 separately (10 mice for each dose). Two weeks after the immunization, in vivo CTL assay and in- tracellular staining (ICS) of IFN-γ were carried out to analyze the dose-effect relationship. Tumor growth and vitiligo (as an sign of autoimmunity) were observed until 3 months after challenge with 105 B I6F10 tumor cells. The results showed that Ad encoding AdmTrp2 induced weak tumor im- mune response. Similar immunization with AdhTrp-2 elicited stronger protective immunity. CTL activity and IFN-γ-produced CD8+T cells were directly proportional to dose of AdhTrp2 or AdmTrp2. Moreover, AdhTrp2 group showed tumor rejection in 100% of challenged mice till the end of 3rd month while 60% of mice immunized with AdmTrp2 were protected against tumor. In the whole process of this experiment, no vitiligo was observed in mice immunized either with AdhTrp2 or AdmTrp2. It is concluded that anti-melanoma responses induced by genetic vaccina- tion expressing xenoantigens breaks immune tolerance effectively and is able to elicit strong anti-gen-specific cytotoxic T cell response without vitiligo.