Objective: This study employed next-generation sequencing (NGS) for large-scale genotyping of 42 blood group systems in blood donors to evaluate its applicability in multi-system blood group identification and rare blood type repository construction, while exploring the complementary value of third-generation sequencing (TGS) in haplotype phasing and variant capture in regions with insufficient sequencing depth for ABO ambiguous samples. Methods: A total of 562 regular blood donors (median donation frequency 11 times) from a blood center were enrolled. NGS was used for genotyping of 42 blood group systems, with a focused analysis on 12 systems essential for rare blood type repository construction: MNS, Lutheran, Kell, Duffy, Kidd, Diego, Yt, Colton, Gerbich, Ok, H, and I. TGS was employed to resolve haplotype phasing (i. e., determining whether different mutation sites are located on the same chromosome) in 12 ABO samples with discrepancies between forward and reverse typing. Serological and genotyping results were compared for MNS, Duffy, Kidd, Lewis, and ABO systems. Results: NGS-based genotyping revealed that Fy (a-b+) in the Duffy system accounted for 0.87% (4/458), and Di (a+b+) in the Diego system accounted for 10.62% (31/292). The Lutheran, Kell, Yt, Colton, H, Ok, I, and Gerbich systems exhibited near-uniform phenotype distributions. Among the 12 ABO ambiguous samples, NGS yielded multiple possible genotype combinations in 5 cases due to inability to phase alleles, whereas TGS uniquely resolved the genotypes through long-read sequencing. In one case, NGS detected only 2 mutation sites due to insufficient sequencing depth, while TGS identified all 11 sites and assigned the A1 phenotype. Concordance rates between serology and genotyping were: Duffy 96.55% (140/145), ABO 96.43% (513/532), Kidd 94.17% (97/103), MNS 85.07% (57/67), and Lewis 68.18% (75/110). In the MNS system, 7 of 8 samples serologically typed as M+N+ but genotyped as M-N+ carried GYPB variants. Lewis discrepancies predominantly featured genotype Le (a-b+) with serological phenotypes of Le (a+b-), Le (a+b+), or Le (a-b-). The NGS platform completed sequencing of 192 samples within 2 weeks. Conclusion: The tiered genotyping strategy combining NGS, TGS, and serology enables multi-system blood group identification in large-scale blood donor populations. TGS provides complementary value in phasing ambiguous ABO samples and detecting variants in regions with insufficient sequencing depth. This study provides a technical framework and baseline frequency data for the expansion of a local rare blood type repository. However, standardization and cost-effectiveness of this strategy require further validation with expanded sample sizes, and serological confirmation should be retained for secretion status-related systems such as Lewis.

Objective: To explore the association between waist circumference and beverage consumption behaviors among junior high school students, so as to provide a theoretical basis for improving their nutritional status. Methods: Using a multi stage cluster random sampling method, 1 611 junior high school students aged 13-15 years from Yushan, Yanshan, Yiyang, and Wannian counties in Shangrao City, Jiangxi Province, were selected for a survey in September 2021 through questionnaires and waist circumference measurements. Stratified analyses were conducted by sex, t-test and analysis of variance (ANOVA) were used to compare waist circumference across groups, and analysis of covariance (ANCOVA) was applied to examine the associations between different types of beverage consumption and waist circumference. Results: Boarding students had significantly greater average waist circumference than day students[boys:(71.86±9.24, 69.69±12.19)cm；girls:(66.53±13.30, 64.63±11.90)cm； t =2.69, 2.08, both P < 0.05 ]. Among boys, waist circumference differed significantly by vigorous physical[vigorous physical:(69.71±9.53)cm; non vigorous exercise:(71.53±12.99)cm]( t=-2.31, P <0.05). Boys waist circumference also differed significantly by consumption frequency (high and medium,low,none) of milk containing beverages[(71.83±7.40, 69.92±9.98, 72.29±13.90)cm], functional beverages[( 73.36± 11.87, 65.97±10.46, 70.71±10.55)cm] and purified water beverages[(69.36±10.87, 71.93±12.15, 71.50±10.09)cm]( F =4.06, 5.93, 4.85, all P <0.05). Girls waist circumference also differed significantly by consumption frequency (high and medium,low,none) of milk containing beverages[(68.81±5.55, 66.59±13.02, 65.36±11.69)cm, F=9.67, P <0.01]. After controlling for the confounding variables, ANCOVA results indicated that the consumption of milk containing beverages, functional beverages and Purified water beverages were significantly associated with waist circumference of boys ( F =3.79, 5.26, 5.08, all P < 0.05 ). Milk containing beverages were significantly associated with waist circumference of girls ( F=4.17, P <0.05). Conclusion The consumption frequency of milk containing beverages and functional beverages by junior high school students is positively correlated with their waist circumference.

[Objective] To study the relationship between the expression intensity of HLA-Ⅰ platelet antibodies in patients with platelet transfusion refractoriness (PTR) and platelet cross-matching, and to further evaluate other factors in order to provide relevant data support for improving platelet transfusion efficiency and optimizing platelet transfusion regimens. [Methods] Luminex single antigen flow cytometry was used to detect platelet specific antibodies in 35 patients with hematological disease. Subsequently, the Capture-P method was employed to perform 102 crossmatchings between plasma with HLA-Ⅰ antibodies and platelets with known HLA-Ⅰ genotypes. The cross-matching results were assessed and the clinical transfusion outcomes were tracked. [Results] The positive detection rate of HLA-Ⅰ and HPA antibodies in this study was 48.6% (17/35). The negative rate of cross-matching in 102 cases was 37.3% (38/102). Multiple factors affect platelet cross-matching, such as HLA-Ⅰ antibody expression level and antibody type, antigen expression level, cross-reactivity group and eplets. Among them, the expression level and antibody type of HLA-Ⅰ antibody are the main influencing factors. However, the effectiveness of clinical platelet transfusion is not completely determined by the compatibility of platelet cross-matching. [Conclusion] In addition to avoiding strong positive HLA-Ⅰ antibodies, clinical matching should also be vigilant against the serological cross-incompatibility caused by weak positive HLA-Ⅰ antibodies. It may be necessary to establish HLA-Ⅰ low expression antigen database as a better alternative platelet donor selection strategy, and gradually explore the effectiveness of ‘low expression mismatch’ strategy for clinical platelet transfusion.

Objective To investigate the diagnostic value of dynamic contrast-enhanced magnetic resonance imaging(DCE-MRI)parameters for cognitive dysfunction in first-episode stroke,thereby providing reference for the early clinical assessment of the risk of cognitive dysfunction in stroke and formulation of intervention programs.Methods A total of 122 patients with first-episode stroke were enrolled and divided into occurrence group(n=53)and non-occurrence group(n=69)according to whether they had cognitive dysfunction.Additionally,53 patients with non-cerebrovascular cognitive dysfunction were selected as control group.All patients underwent DCE-MRI examination after admission.The 3 groups and patients with different degrees of cognitive dysfunction were compared for DCE-MRI parameters,including interstitium-to-plasma rate constant(Kep),volume transfer constant(Ktrans),volume fraction of extravascular extracellular space(Ve),and volume fraction of plasma(Vp).The correlations of DCE-MRI parameters with the degree of cognitive dysfunction were analyzed,along with their diagnostic value for cognitive dysfunction and evaluation value for the risk of cognitive dysfunction after first-episode stroke.Results Both occurrence group and non-occurrence group had higher levels of Ktrans,Ve and Vp than control group,and these parameters were higher in occurrence group than non-occurrence group(P<0.05).Ktrans,Ve and Vp of patients with different degrees of cognitive dysfunction differed significantly(P<0.05),and they were increased significantly with the aggravation of cognitive dysfunction.Ktrans,Ve and Vp were all positively correlated with the degree of cognitive dysfunction (P<0.05). The AUC for the combined diagnosis using Ktrans, Ve and Vp was 0.921, significantly higher than single index (P<0.05). The risks of cognitive dysfunction in first-episode stroke patients with higher levels of Ktrans, Ve and Vp were 3.077, 1.944 and 2.313 times of the low-level, respectively. Conclusion DCE-MRI can be used to diagnose cognitive dysfunction in first-episode stroke, providing reference for early clinical prediction of cognitive dysfunction after stroke.

Humans ; COVID-19/genetics* ; SARS-CoV-2/genetics* ; Clustered Regularly Interspaced Short Palindromic Repeats/genetics* ; CRISPR-Cas Systems/genetics* ; RNA, Viral/genetics*

Objective To investigate the infection of human parvovirus B19 in Suzhou voluntary blood donors under the current blood screening model. Methods A total of 893 blood donor samples from September to December 2022 were randomly collected. Samples were tested to determine the seroprevalence (anti-B19 IgG and IgM) of B19 antibodies by enzyme-linked immunosorbent assay(ELISA), and B19 DNA of positive samples was further detected by real-time polymerase chain reaction(PCR) assay. Results Among 893 samples, the total seroprevalence of B19 antibody was 20.7% (185/893), with anti-B19 IgG and IgM positive rate at 19.4% (173/893) and 1.9% (17/893), respectively, showing significant difference (P<0.05). No difference in the positive rates of B19 IgG and IgM (20.1%, 1.5% vs 18.0%,2.6%) was noticed by gender(P>0.05). The prevalence of anti-B19 IgG statistically increased with age (P<0.05), while there was no difference in the prevalence of anti-B19 IgM (P>0.05). No statistical difference was not found in anti-B19 IgG and IgM samples among different blood groups. The anti-B19 IgG in repeated blood donors was higher than that in first-time donors(21.5% vs 15.9%)(P<0.05) while there was no difference in the positive rate of IgM antibodies (P>0.05). Three cases were found to be positive for B19 DNA in the B19 antibody positive samples, with the positive rate at 1.6%(3/185). Conclusion Although the prevalence of B19 infection in Suzhou was lower than that in other areas and was mostly past infection, there was still a certain proportion of persistent infection and acute infection, which posed the potential risk of blood transfusion transmission. Therefore, attention should be paid to blood transfusions, especially for the high-risk and susceptible groups.

【Objective】 To analyze differences of eplets between the patient who generated HLA allele-specific antibodies after platelet transfusion with donors. 【Methods】 The HLA genotypes of the patient and donors were detected by PCR-SBT, and the Luminex single antigen beads coating was used to screen HLA-Ⅰ antibodies in the patient’s serum. HLA Matchmaker was utilized to analyze different amino acids and eplets. 【Results】 The patient carried HLA-A^*02∶03 allele, and HLA-A2 antibodies were found in his serum after platelet transfusion (A^*02∶01, A^*02∶06, and A^*02∶07). Sequence alignment showed that the patient′s A^*02∶03 has a difference in position 149, which resulted in a different eplet between A^*02∶03 and A^*02∶01, A^*02∶06, A^*02∶07 and then induced the production of antibodies. 【Conclusion】 HLA antibodies are specific for HLA epitopes that have structural differences due to amino acid differences between HLA alleles, suggesting that high-resolution typing of HLA-A, -B need to be conducted in patients and donors, and the acceptable mismatch of HLA should be determined based on epitopes rather than antigens, so as to reduce alloimmune response and improve platelet count after transfusion.

【Objective】 To study the incidence and specificity of platelet antibody in blood donors in Suzhou, analyze the distribution characteristics of platelet antibody in blood donors in this area, and explore the significance of platelet antibody detection in blood donors to reduce the adverse reactions toplatelet transfusion in clinical. 【Methods】 Platelet antibody detection was performed in 2178 blood donors in this area by solid-phase immunosorbent assay. The antibody specificity of the positive samples was analyzed by commercial kit, and the anti-CD36 antibody positive samples were further identified by flow cytometry and gene sequencing. 【Results】 Twelve positive samples were detected by platelet antibody screening, with a positive rate of 0.55%(12/2 178), including 5 males (0.33%, 5/2 178)and 7 females(1.06%, 7/2 178). Among the positive samples, anti-HLA-Ⅰ antibody was identified in 2 cases, anti-CD36 antibody in 1 case, and the antibody specificity was not identified in the other 9 cases. In one case, the positive rate of anti-HLA-Ⅰ antibody PRA was 31.31%(31/ 99), which was mainly specific to anti-B15, anti-B35 and anti-B40. The positive rate of anti-HLA-Ⅰ antibody PRA in the other case was 45.45%(45/ 99), which was mainly specific to anti-A2, anti-A11, anti-A24, anti-A29, anti-A33, anti-A66, anti-B15 and anti-B35. The blood donor with anti-CD36 antibody was type I CD36 deficiency, and 329_330delAC mutation occurred in exon 5. 【Conclusion】 Through antibody screening and specificity identification, the positive rate of platelet antibody in females was significantly higher than that in males(P<0.05). In addition to the common anti-HLA-I antibodies, anti-CD36 antibody was also detected in type I CD36 deficient blood donor. Therefore, the detection of platelet antibodies in blood donors is of certain clinical significance to reduce the adverse reactions to blood transfusion caused by antibodies in platelet products.

【Objective】 To investigate the expression of CD36 antigen in Suzhou area, analyze the type of antigen deficiency and gene mutation, so as to provide references for the establishment of CD36 negative donor registry in Suzhou. 【Methods】 Anticoagulant whole blood samples (805 cases) were randomly collected from healthy blood donors in Suzhou Blood Center. The expression of CD36 antigen on platelet and monocyte was analyzed by flow cytometry to determine the type of CD36 deficiency. The gene mutation type of platelet CD36 antigen-deficient was performed by genomic DNA sequencing. 【Results】 The CD36 deficiency frequency on platelet was 2.48% (20/805), among which TypeⅠ(lacking CD36 expression both on platelet and monocyte) and TypeⅡ(lacking CD36 expression on platelet only) CD36 deficiency accounted for 10% (2/20) and 90% (18/20), respectively. CD36 gene mutations were found in 10 samples, including 3 cases of 329_330 d^elAC, 1 case of 1228_1239 d^elATTGTGCCTATT and 2 cases of 1163 A>T; 1 case of 329_330 d^elAC+ 1172_1183 d^elTATTGGTCAAGC and 287 G>C+ 329_330 d^elAC heterozygous mutation. In addition, 1 case of 745 A>G and 1 case of 806 C>T mutations were novel, and not yet reported. 【Conclusion】 Results showed that the frequency of CD36 antigen deficiency in Suzhou were similar to that reported in southern China, but the mutation sites were slightly different. The establishment of CD36 negative platelet registry could provide negative platelets for patients with transfusion reactions caused by anti-CD36 antibody and improve the effect of clinical platelet transfusion.

Objective To compare the short-term efficacy of AC TightRope plate and clavicular hook plate in the treatment of acromioclavicular joint dislocation.Methods A retrospective case control study was conducted to analyze the clinical data of 32 patients with acromioclavicular joint dislocation admitted to Yifu Hospital Affiliated to Nanjing Medical University from April 2016 to December 2017.There were 23 males and 9 females,aged 26-61 years [(43.5 ±9.5)years].According to Rockwood classification,there were 23 patients with type Ⅲ and 9 patients with type Ⅴ.The patients were divided into AC TightRope group and clavicular hook plate group according to different treatment methods.There were 17 patients in AC TightRope group,including 12 males and 5 females,aged (42.6 ± 10.3)years.There were 15 patients in clavicular hook plate group,including 11 males and 4 females,aged (44.5 ± 8.9)years.The incision length,operation time,intraoperative blood loss,hospitalization time,postoperative visual analogue scale (VAS) of the affected shoulder,functional recovery of shoulder joint and complications were compared between the two groups.Results All patients were followed up for 6-26 months,with an average of 15 months.There was no significant difference between the two groups in hospitalization time (P ＞ 0.05).In the AC TightRope group and clavicular hook plate group,the operation time was (41.5 ±4.2)minutes and (60.1 ± 4.2) minutes,respectively (P ＜ 0.01);the incision length was (3.6 ± 0.3) cm and (10.1 ± 0.6) cm,respectively (P ＜ 0.01);the intraoperative blood loss was (45.3 ± 11.1) ml and (78.3 ± 16.0) ml,respectively (P ＜ 0.01);the postoperative VAS of the shoulder was (1.9 ± 0.5) points and (3.1 ± 0.7) points,respectively (P ＜ 0.05);the excellent and good rate of postoperative recovery of shoulder joint function was 100％ (17/17) and 67％ (10/15),respectively (P ＜0.01).There were no complications such as surgical incision infection,intraoperative neurovascular injury or condylar fracture during the perioperative period.No serious postoperative complications such as internal fixation loosening,loss of reduction or recurrence of dislocation were found in both groups.Conclusion For acromioclavicular joint dislocation,AC TightRope can obtain good clinical effect and has obvious advantages in operation time,incision length and blood loss,with lower incidence of shoulder pain and better functional recovery compared with clavicular hook plate.