[Objective] To analyse the distribution of antigen phenotypes in the Rh, MNS and Kidd blood group systems of Han and Tujia blood donors in Chongqing, and to provide data support for the establishment of an expanded blood group antigen phenotype database and the development of expanded blood group coordinated transfusion in blood donors. [Methods] The antigens of Rh, MNS and Kidd blood group systems in Han and Tujia blood donors in Chongqing were detected by test-tube method, and the Hardy-Weinborg anastomosis of the three blood group systems was calculated. Pearson's chi-square test and Fisher's exact probability method were used to compare the differences in phenotypic distribution frequencies among different regions and ethnic groups. [Results] Han and Tujia blood donors accounted for the highest proportion of CCee in the antigenic phenotype of the Rh blood group system, followed by CcEe, and then Ccee and ccEE. Tujia blood donors accounted for 52.02% of CCee, which was higher than that of Han blood donors (47.24%), while Han blood donors accounted for 32.20% of CcEe, which was higher than that of Tujia blood donors (28.94%). In the antigenic phenotype of the MNS blood group system, the blood donors of Han nationality and Tujia were MN>MM>NN,. The antigen phenotype distribution frequency of the Kidd blood group system was highest for Jk(a+b+) among both Han and Tujia blood donors, and the blood donors of Han nationality were Jk(a+b+)>Jk(a+b+), while those of Tujia were Jk(a-b+)>Jk(a+b-). The antigens of the three blood groups of Han and Tujia blood donors were consistent with the Hardy-Weinberg balance(P>0.05). There was no statistically significant difference in the frequency of antigen phenotypes of the three blood group systems between Han and Tujia blood donors(P>0.05). There were statistically significant differences in the phenotypic distribution frequency of Rh antigens between Chongqing and Xi'an, Zhejiang, Shantou, Foshan, Nanning and Yangzhou(P<0.05), but not with Guang'an and Shenzhen(P>0.05). There were statistically significant differences in the phenotypic distribution frequency of Rh antigens between Han, Tujia, Zang, Mongolian, Korean and Hani ethnic groups in Chongqing(P<0.05). There were statistically significant differences in the phenotypic distribution frequency of MNS antigens between Han blood donors in Chongqing and Urumqi, Hainan and Yuncheng, but not with Xi'an and Wenzhou. There was a statistically significant difference in the phenotypic distribution frequency of MNS antigen between Tujia blood donors in Chongqing and Urumqi and Hainan(P<0.05), but there was no significant difference in the phenotypic distribution frequency of MNS antigen between Tujia blood donors in Chongqing, Urumqi and Hainan(P>0.05). There was a statistically significant difference in the phenotypic distribution frequency of Kidd antigens between blood donors in Chongqing and Harbin(P<0.05), but not in Huizhou, Wenzhou and Yichang(P>0.05). [Conclusion] The population in Chongqing has multi-ethnic characteristics, and the antigenic phenotypes of Rh, MNS and Kidd blood group systems exhibit diversity and regional differences. Establishing an expanded blood bank can provide more options for precision blood transfusion.

Objective: To investigate the clinical characteristics, the types of unexpected antibodies, and their impacts on immunological risks among patients with different frequencies of cross-matching incompatibility, so as to propose corresponding solutions. Methods: Data of cross-matching incompatibility samples from 92 medical institutions during 2022 to 2024 were collected and divided into three groups based on the frequency of cross-matching. Statistical analysis was performed on disease types, distribution of hematologic diseases, alloantibody detection rates, and proportions of alloantibody types. Results: The 858 patients were divided into three groups based on the frequency of blood cross-matching incompatibility: ≥5 times (8.28%, 71/858), 2 to 4 times (28.21%, 242/858); 1 time (63.52%, 545/858). There was a clustered distribution of disease types in the ≥5 cross-matchings group, with 71.83% (51/71) of patients having tumors or hematologic and hematopoietic diseases. In contrast, the disease types in the 2 to 4 cross-matchings and 1 cross-matching groups were more diverse. An analysis of 249 patients with hematologic diseases found that multiple myeloma was the most common disease in all three groups, accounting for 31.43% (11/35), 35.37% (29/82), and 37.88% (50/132) respectively. In the ≥5 cross-matchings group, myelodysplastic syndrome (14.29%, 5/35) and thalassemia (14.29%, 5/35) were the second most common diseases. In contrast, in the 2 to 4 cross-matchings group and 1 cross-matching group, autoimmune hemolytic anemia was the second most common disease, with prevalence rates of 20.73% (17/82) and 24.24% (32/132), respectively. Alloantibodies were detected in 54.66% of the patients, with antibodies against Rh blood group being most frequent (>50%) in all three groups. The detection rates of alloantibodies/alloantibodies with coexisting autoantibodies decreased across groups: the ≥5 cross-matchings group (70.42%, 50/71) > the 2 to 4 cross-matchings group (54.96%, 133/242) > the 1 cross-matching group (52.48%, 286/545). Conclusion: The risk of alloantibody production increases in patients with multiple cross-matching incompatibilities, especially in those with tumors or hematologic diseases. For handling of cross-matching incompatibility cases, it is recommended to optimize the cross-matching process, implement individualized transfusion plans, and enhance the technical capabilities of clinical transfusion departments and blood group reference laboratories to ensure the safety and effectiveness of transfusions.

Objective To investigate the risk factors of postoperative fecal contamination in children pa-tients with Hirschsprung's disease(HSCR),and to construct and evaluate the risk predictive model.Methods The clinical data in 377 children patients with HSCR in 3 class 3A hospitals in Guangxi from Janu-ary 2016 to June 2021were retrospectively analyzed by adopting the convenience sampling method.The pa-tients were divided into the modeling group(n=264)and testing model group(n=113)with a ratio of 7∶3.The risk factors of postoperative fecal soiling were analyzed by the single factor and multiple factors,and the risk predictive model was constructed.The receiver operating characteristic(ROC)curve was used to detect the discriminative ability of the model and the H-L test was used to determine the goodness of fit of the mod-el.The model was prospectively validated in 21 children patients with HSCR from August to December 2021.Results Among 377 children patients with HSCR,the fecal soiling occurred in 131 cases with a incidence rate of 34.75％.The constructed predictive model of fecal contamination risk after HSCR operation:logit(P)=-2.385+1.697 × special type of megacolon+0.929 × Soave+0.105 × length of bowel resection+2.065 × il-literate caregivers+0.808 × caregivers'implementation of postoperative diet+0.867 × postoperative defecation training by caregivers.The area under the curve(AUC)in the modeling group was 0.849,the Yoden index was 0.53,the optimal critical value of the model was 0.32,the sensitivity was 76.00％,and the specificity was 77.00％.The H-L test,X2=6.649,P=0.575.AUC of the testing model group was 0.736,the sensitivity was 81.25％,and the specificity was 78.46％.The prospective validation results showed that the sensitivity and specificity of the model were 66.67％and 100％respectively.Conclusion The constructed model has good i-dentification and predictive ability.

Objective: To investigate the causal relationship between antioxidant nutrients and pregnancy complications, so as to provide the reference for the prevention and treatment of pregnancy complications. Methods: Data of seven antioxidant nutrients including vitamin A, vitamin C, vitamin E, selenium, zinc, copper and iron were collected from genome-wide association study (GWAS) Catalog database, and data of four pregnancy complications including gestational diabetes mellitus, pre-eclampsia, spontaneous abortion and preterm birth were collected from the Finland database. Single nucleotide polymorphism (SNP) data were collected, and 27 SNPS strongly correlated with seven antioxidant nutrients were selected as instrumental variables. Mendelian randomization (MR) analyses were performed using the inverse-variance weighted (IVW) method with seven antioxidant nutrients as exposures factors and four pregnancy complications as outcome variables. The heterogeneity was assessed using the Cochran's Q test, the horizontal pleiotropy was assessed using the MR-PRESSO test and MR-Egger regression, and the robustness of the results was verified with the leave-one-out. Results: Cochran's Q test showed heterogeneity of MR results between vitamin C and gestational diabetes mellitus, preeclampsia and preterm birth, between vitamin E and iron and gestational diabetes (all P<0.05), and a random effect model was employed. There was no heterogeneity in other results (all P>0.05), and a fixed effect model was employed. MR analysis results showed that there was no causal association between seven antioxidant nutrients and the risk of four pregnancy complications (all P>0.05). MR-PRESSO test and the MR-Egger regression identified no horizontal pleiotropy of instrumental variables (both P>0.05). Conclusion This study did not find genetically predicted associations of antioxidant nutrients with pregnancy complications.

【Objective】 This study endeavors to introduce the statistical process control (SPC) method to analyze the quality control index concerning red blood cells in additive solution with leukocytes reduced, with the aspiration to advance the effective utilization of blood quality control data, thereby providing empirical foundations for the continual enhancement of blood quality. 【Methods】 Between 2020 and 2022, test data pertaining to the quality control index of red blood cells in additive solution with leukocytes reduced were amassed from six blood stations in Chongqing area. Utilizing Minitab software, the SPC analysis was carried out, p-control charts were delineated, the non-conformance rates of each quality control index along with their 95% confidence intervals were computed, as well as the Process Capability Index (Z value). 【Results】 In accordance with the Whole Blood and Blood Components Quality Requirements, the appraisal of the quality control indexes for red blood cells in additive solution with leukocytes reduced manifested a conformity rate of 100% for appearance, end-of-storage hemolysis rate and sterility test. Nonetheless, the conformity rates for volume, hemoglobin, hematocrit and residual leukocytes did not attain 100%, albeit all were ≥75%. Through the employment of binomial distribution-based p-control charts, the controlled state of the production process was discerned. Although the overarching conformity rate satisfied the national standard stipulations, it was discerned that there were out-of-control points concerning volume, hemoglobin, hematocrit, and residual leukocytes across different institutions, exhibiting palpable trends. The non-conformance rates of all quality control indexes were less than 25%, yet at a 95% confidence level, the residual leukocyte counts from institutions B, C, E, and F did not adhere to the stipulations (exceeding 25%). By architecting the ability evaluation index Z value for count data process capability analysis, it was unveiled that the volume of institution E, the hematocrit of institutions B, C, and F, and the residual leukocytes Z values of all six blood collection and supply institutions were below 2, hinting at avenues for amelioration. 【Conclusion】 The SPC method anchored in binomial distribution exhibits substantial application merit in blood component quality management, facilitating real-time surveillance of blood collection, preparation, and storage procedures.

Objective To explore the application value of ZZAP reagent in treating allogeneic erythro-cytes for absorption experiment use and in excluding the interference of erythrocyte autoantibodies in pre-transfusion tests.Methods The serological characteristics of 413 cases of erythrocyte autoantibodies interfer-ence in pre-transfusion tests in this center from January 2017 to February 2024 was retrospectively analyzed.The antibody identification adopted different methods.The patients were divided into 3 groups according to the antibody identification method:self-absorption group(the serum after autologous cell absorption test con-ducted the antibody identification and cross matching,n=87),unabsorption group(the serum of the patients conducted the antibody identification and cross-matched,n=277)and allogenic absorption group(the serum after allogeneic erythrocyte absorption test treated by ZZAP reagent condcuted the antibody identification and cross-matched,n=49).Among them,77 cases of allogenic absorption treated by ZZAP reagent and without absorption test were selected and conduct the detailed survey on the blood infusion effective rate and iso-anti-body detection situation.The changes of red blood cell(RBC)count,hemoglobin(Hb)and hematocrit(Hct)were compared before and after blood transfusion.The effect of allogenic cells absorption method for exclu-ding the autoantibodies interference and increasing the blood transfusion effect after ZZAP reagent treating al-logenic cells was analyzed.Results The typing results of antihuman globulin test(direct method)in the pa-tients with RBC atoantibodies were mainly IgG positive and IgG+C3d positive,and the autoantibodies were mainly the warm autoantibodies and cold and warm mixed autoantibodies.HB,RBC Hct after blood transfu-sion in the allogenic absorption group and unabsorption group all were improved.The alloantibody detection rate in the allogenic adsorption group was 42.86％,which was significantly higher than 12.64％in the unab-sorbed group and 11.49％in the autologous adsorption group,and the differences were statistically significant(P＜0.001).The total effective rate of blood transfusion in the allogenic adsorption group was 95.92％,which was significantly higher than 78.57％in the unabsorbtion group,and the differences were statistically significant(P=0.016).Conclusion The adsorption of allogeneic cells after ZZAP treatment could serve as a substitute scheme for the inoperable autoadsorption,which effectively excludes the interference of autoanti-bodies on the pre-transfusion test,and increase the safety and effectiveness of blood transfusion.

LC3-associated phagocytosis (LAP) is a special phagocytosis occurring at the intersection of the two pathways of phagocytosis and autophagy. A hallmark event of the LAP process is the recruitment of microtubule-associated proteinⅠlight chain type 3-Ⅱ(LC3Ⅱ) to the phagosome surface of the monolayer membrane structure. The LAP pathway relies on the functions of the RUN domain and cysteine-rich domain containing, Beclin 1-interacting protein (Rubicon) and reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. The LC3-associated phagosome (LAPosome) binds to the lysosome to digest and degrade the contents. In recent years, increasing studies have found that LAP plays an important role in the infections caused by pathogenic microorganisms including fungi and bacteria. LAP is a crucial way in the host to resist and degrade the infection of pathogenic microorganisms. However, some pathogenic microorganisms can effectively escape from LAP in the host and even use LAPosome as a place for colonization and replication. This article summarized the recent progress in the role of LAP in host defense against pathogenic microorganism infection and the significance of it in the occurrence and development of diseases.

Objective:To analyze the clinical and molecular genetic characteristics of a Chinese family with congenital cataract-microcornea syndrome.Methods:The method of pedigree investigation was adopted.A Chinese Han family with congenital cataract-microcornea syndrome was recruited in Xiamen Eye Center of Xiamen University.All the family members received detailed ophthalmologic examination including the best corrected visual acuity, intraocular pressure measurement by handheld applanation tonometry, slit lamp biomicroscopy, color fundus photography, B-scan ultrasonography, corneal diameter, anterior segment optical coherence tomography, ultrasound biomicroscopy, corneal endoscopy, and corneal topography.Genomic DNA was extracted from peripheral venous blood from some patients and unaffected family members.Targeted high-throughput DNA sequencing was performed on the proband.The sequencing chip contained 188 known pathogenic genes related to lens abnormalities.Suspected pathogenic genes were verified by Sanger sequencing in phenotypically normal family members to identify the co-segregation and the disease-causing gene.Bioinformatics analysis was performed to analyze the pathogenicity of variants by REVEL.Conserved protein domains were analyzed by InterPro.Physicochemical property of the mutant protein was analyzed by ProtParam.The deleteriousness of the protein was predicted by PolyPhen-2.Homology of the variants in pathogenic gene was analyzed by NCBI website to compare the conservation among various species.This study followed the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Xiamen Eye Center of Xiamen University (No.XMYKZX-LW-2009-003).Written informed consent was obtained from each subject prior to entering the study cohort.Results:There were 39 members of 4 generations in this family including 11 patients with an autosomal dominant inheritance pattern.Clinical features of the patients included congenital cataract and microcornea.No obvious abnormality was found in ophthalmic and general examination.A heterozygous mutation c. 61C>T in the CRYAA gene was found, resulting in the mutation of the amino acid from arginine to tryptophan (p.Arg21Trp) at position 21, consistent with co-segregation.The number of cationic cluster in the mutant protein decreased, and the hydrophilicity and stability were reduced.The variant was predicted to be deleterious and was highly conserved in multiple species. Conclusions:A novel heterozygous mutation c.61C>T p. Arg21Trp in CRYAA gene is considered as the causal gene of this family.It is the first time this variant has been reported in China.

Objective:To investigate the role of NOD-like receptor protein 3 (NLRP3) in the regulation of phagocytosis in Vibrio vulnificus ( V. vulnificus)-infected macrophages. Methods:Expression profiles of phagocytosis-related genes in PBS- and V. vulnificus-infected J774A.1 cells were analyzed by RNA-Seq. NLRP3-knockout (NLRP3 KO) J774A.1 cells were constructed using CRISPR-Cas9 gene-editing system. The phagocytosis of V. vulnificus and pHrodo RED-labelled Escherichia coli ( E. coli) bioparticles in parental and NLRP3 KO J774A.1 cells was detected by flow cytometry. Real-time PCR was performed to measure the expression of Fgr2 b gene at mRNA level in PBS- and V. vulnificus-treated parental and NLRP3 KO J774A.1 cells. Results:The expression of 18 phagocytosis-related genes was upregulated in V. vulnificus-infected J774A.1 cells than in PBS-treated J774A.1 cells ( P<0.05). There was a 5 bp deletion in the exon 2 of NLRP3 gene in NLRP3 KO J774A.1 cells, resulting in frameshift mutation and complete loss of NLRP3 expression. NLRP3 KO J774A.1 cells exhibited enhanced phagocytosis of V. vulnificus and pHrodo RED-labelled E. coli bioparticles than parental J774A.1 cells ( P<0.05). Besides, the expression of Fgr2 b gene at mRNA level was significantly increased in V. vulnificus-infected NLRP3 KO J774A.1 cells than in parental J774A.1 cells ( P<0.05). Conclusions:The phagocytosis of V. vulnificus in macrophages could be negatively regulated by NLRP3, which was possibly mediated through the regulation of Fgr2 b gene expression.

Objective:To investigate the effects of TNF-α knockout on liver and spleen neutrophil responses to Vibrio vulnificus bloodstream infection in a mouse model. Methods:(1) TNF-α-knockout (TNF-α -/-) and wild-type (WT) C57BL/6J mice aged 6-8 weeks were randomly divided into four groups with six in each group: uninfected WT group, infected WT group, uninfected TNF-α -/- group and infected TNF-α -/- group. The mouse model of bloodstream infection was constructed by intraperitoneal injection of Vibrio vulnificus CGMCC1.1758 (2×10 8 CFU/200 μl), while the mice in the uninfected groups were injected intraperitoneally with equal amount of PBS. (2) Liver immune cells and splenocytes were isolated 4 h after infection and subjected to analyze the percentages and numbers of neutrophils, and the changes in cell viability, cellular reactive oxygen species (ROS) level and phagocytosis by flow cytometry. In addition, effects of Vibrio vulnificus bloodstream infection on mTOR signaling pathway in murine neutrophils were evaluated in vivo. Results:(1)Compared with the uninfected WT group, the percentages and numbers of neutrophils in liver and spleen tissues of the infected WT group increased significantly. The percentage and number of liver neutrophils were significantly higher in the infected TNF-α -/- group than in the infected WT group, but no significant difference in spleen neutrophils was detected between the two groups. (2) Compared with the infected WT group, the phagocytosis of liver neutrophils rather than that of spleen neutrophils was enhanced in the infected TNF-α -/- group. (3) The survival rates of neutrophils in both liver and spleen were decreased, while the cellular ROS level was significantly increased in the infected WT group compared with those of the uninfected WT group. Compared with the infected WT group, the infected TNF-α -/- group had increased survival rates of both liver and spleen neutrophils, but decreased level of ROS. (4) The levels of p-AKT (S473) in liver and spleen neutrophils of the infected WT group were lower than those of the uninfected WT group. Compared with the infected WT group, the infected TNF-α -/- group had lower level of p-AKT (S473) in liver neutrophils, but higher p-AKT (S473) level in spleen neutrophils. There were no significant differences in p-4E-BP1(T37/46) levels between the uninfected WT group and the infected WT group. The p-4E-BP1 (T37/46) level in liver neutrophils was lower in the infected TNF-α -/- group than in the infected WT group, but no significant difference in p-4E-BP1 (T37/46) levels in spleen neutrophils was observed between the two groups. Conclusions:TNF-α had different effects on the neutrophils in spleen and liver tissues of mice with Vibrio vulnificus bloodstream infection. It played a critical role in regulating the recruitment, phagocytic function and mTOR signaling of liver neutrophils after Vibrio vulnificus infection in vivo.