1.Studies on the effect of the p.Arg352Gln mutation on Bw07 transferase
Yongkui KONG ; Pengfei ZHU ; Xin LIU ; Jie SONG ; Li WANG ; Shuya WANG ; Qiankun YANG
Chinese Journal of Laboratory Medicine 2024;47(3):286-292
Objective:To investigate the molecular biological mechanism of Bw07 allele and its transferase alteration carried by a proband of ABw07 subtype.Methods:A 2-year-old male child was selected as the research object. The peripheral blood of the proband and his parents was identified for ABO blood type by the test tube method, and the ABO subgroup PCR-SSP detection and ABO gene sequencing were performed on the three individuals to determine their blood type genotypes. Finally, the effect of the p.Arg352Gln mutation on Bw07 transferase was verified by virtual mutation, DUET structure prediction, molecular dynamics analysis, and in vitro cellular experiments.Results:The serological phenotypes of the proband and his mother were ABw and Bw, respectively, while his father was normal A. The ABO subgroup PCR-SSP assay identified the three genotypes as Bw07/A, Bw07/O, and A/A, respectively.Sanger sequencing further verified that the proband and his mother carried the Bw07 gene, and virtual mutation showed that the intermolecular forces were weakened by the R352Q mutation. DUET predicted that this p.Arg352Gln mutation could affect the thermodynamic stability of Bw07 transferase. Molecular dynamics analysis confirmed that the alteration of thermodynamic stability was mainly related to the appearance of large fluctuations in the amino acid backbone atoms in the 125-133, 193-198 and 336-354 regions, and in vitro cellular experiments further verified the weakened antigen synthesis of Bw07 transferase.Conclusion:The formation of the ABw07 phenotype is associated with the mutation of the highly conserved Arg352 to Gln in Bw07 transferase.
2.Changes in perioperative blood group antibody of 33 type-A/B recipients in ABO-incompatible kidney transplanta-tion
Huifang JIN ; Yongkui KONG ; Xin LIU ; Shuya WANG ; Liyinghui CHEN ; Hao YANG ; Jinfeng LI ; Qiankun YANG
Chinese Journal of Blood Transfusion 2024;37(5):534-540
Objective To statistically analyze the perioperative results of patients with ABO-incompatible kidney trans-plantation(ABOi-KT),in order to explore the changes in blood group antibody of type-A/B recipients.Methods A total of 33 cases of blood group A/B ABOi-KT recipients in our hospital from January 2021 to October 2023 were recruited and divided into two groups of group A(n=18)and group B(n=15)according to the different blood types of recipient.The effects of preoperative plasmapheresis on antibody titer,antibody rebound and renal function after operation(serum urea ni-trogen,creatinine and estimated glomerular filtration rate on the 1st,3rd,7th and 14th day)were analyzed between the two groups.According to the postoperative rebound of blood type antibodies,33 recipients were divided into antibody rebound group(n=7)and non rebound group(n=26),and the differences in initial blood type antibody titers between the two groups were analyzed.Results There was no significant difference in the clearance rate of IgM with preoperative plasma ex-change between the two groups(Z=-0.26,P>0.05);Levels of serum urea nitrogen and creatinine on the 1st,3rd,7th and 14th day after operation between group A and group B were not statistically significant(P>0.05),the same as eGFR.Group B was more prone to rebound antibody compared with group A(P<0.05).There was a significant difference in the in-itial IgM antibody titer between the blood type antibody rebound group and the non rebound group(Z=-2.127,P<0.05),but no statistically significant difference in the initial IgG antibody titer(Z=-1.835,P>0.05)between the two groups was found.Conclusion The patients type B receiving type AB kidney donors are more prone to rebound antibody after ABOi-KT operation compared to the the patients type A receiving type AB.
3.Exploration of the theoretical basis,feasibility,and path of integrating life education into the cultivation of college students'"three views"
Chinese Medical Ethics 2024;37(8):963-966
Life education,as an important modern educational tool,can guide people to view talent cultivation from the dimension of the life cycle.University period is a crucial period for establishing a correct worldview,outlook on life,and values,as well as fastening the first button of life.From the perspective of the ultimate goals,the cultivation of"three views"resonated with the same frequency as life education.Therefore,timely integrating life education into college students'"three views"can not only help promote the innovative development of college students'life education content,but also help college students to correctly understand the world,times,and individuals,complete the shaping of the positive"three views",and shoulder the mission entrusted to youth people by the times.
4.Molecular study of an individual with Bel subtype due to a novel c. 620T>C variant
Xin LIU ; Huifang JIN ; Shuya WANG ; Ying XIE ; Xue LIU ; Yinghui CHEN-LI ; Yongkui KONG
Chinese Journal of Medical Genetics 2024;41(4):411-416
Objective:To explore the molecular basis for an individual with Bel subtype of the ABO blood type due to a novel c. 620T>C variant gene, and assess its impact on the structure of GTB transferase.Methods:An individual who had visited the First Affiliated Hospital of Zhengzhou University on February 11, 2023 was selected as the study subject. ABO phenotyping was initially conducted with serological methods, which was followed by direct sequencing of 7 exons of the ABO gene. Subsequently, single-strand sequencing was carried out by using allele-specific primers, and the variant in the B transferase was homology-modeled using the Modeller software. The impact of the variant on the transferase′s spatial structure was analyzed with the PyMOL software. Results:The serological phenotype of the patient was identified as the Bel subtype. Direct sequencing revealed that she has harbored a novel c. 620T>C variant, resulting in a p. Leu207Pro substitution in the polypeptide chain. Combined with single-strand sequencing, her genotype was ultimately determined as ABO* BELnew/ ABO* O.01.02. Three-dimensional protein structure modeling showed that, compared with the wild type, the distance of one hydrogen bond between Proline and Glycine at position 272 has increased, along with disappearance of another hydrogen bond. Conclusion:The novel c. 620T>C (p.Leu207Pro) variant of B allele may affect the structural stability of the glycosyltransferase. The weakened enzyme activity in turn may lead to reduced B antigen expression, manifesting as the Bel subtype by serological analysis.
5.Molecular biological identification of a case with A223B subtype
Li WANG ; Qiankun YANG ; Shuya WANG ; Ying XIE ; Xue LIU ; Yanli CHANG ; Yongkui KONG
Chinese Journal of Medical Genetics 2024;41(8):982-987
Objective:To study the molecular basis for a proband with A subtype B of the ABO blood group and explore the influence of amino acid variant on the activity of glycosyltransferase (GT).Methods:A proband who had presented at the First Affiliated Hospital of Zhengzhou University on July 2, 2020 was selected as the study subject. Serological identification of the ABO blood groups of the proband and her family members were performed by gel card and test tube methods. The ABO gene of the proband was identified by PCR-sequence specific primers (PCR-SSP) and DNA sequencing. A 3D molecular homologous model was constructed to predict the impact of the variant on the stability of α-(1→3)-D-N-acetylgalactosamine transferase (GTA). Results:The red blood cells of the proband, her mother and two younger brothers showed weak agglutination with anti-A and strong agglutination with anti-B. The sera showed 1~2+ agglutination with Ac and no agglutination with Bc. Based on the serological characteristics, the proband was identified as AwB subtype. Pedigree analysis suggested that the variant was inherited from her mother. The blood group of the proband was identified as A223B type by PCR-SSP. ABO gene sequencing analysis showed that the proband has harbored heterozygous variants of c. 297A>G, c. 467C>T, c. 526C>G, c. 657C>T, c. 703G>A, c. 796C>A, c. 803G>C, c. 930G>A and c. 1055insA. Based on the results of clone sequencing, it was speculated that the genotype was ABO* A223/ ABO* B.01. There were c. 467C>T and c. 1055insA variants compared with ABO* A1.01, and c. 1055insA variant compared with ABO* A1.02. Homologous modeling showed that the C-terminal of A223 GT was significantly prolonged, and the local amino acids and hydrogen bond network have changed. Conclusion:Above results revealed the molecular genetics mechanism of A223B subtype. The c. 1055insA variant carried by the proband may affect the enzymatic activity of GTA and ultimately lead to weakening of A antigen.
6.Characteristics of oral flora and its metabolites in children with henoch-schonlein purpura
Qin-Gwen WANG ; Shuya ZHANG ; Weilin XIONG ; Xiaolei HU ; Ziwei LI ; Qingyin GUO
The Journal of Practical Medicine 2024;40(9):1244-1250
Objective To study and compare the oral microbiota and metabolites of children with Henoch Schonlein purpura(HSP)to identify specific microbiota and metabolites related to this disease and elucidate the pathogenesis of HSP.Methods Three groups of qualified subjects were included,including 20 in the HSP group,20 in the HSP nephritis(HSPN)group,and 20 in the control group.Perform high-throughput 16S rRNA sequencing and metabolic profiling of saliva from each group to analyze the correlation between differential microbiota and differ-ential metabolites.Results(1)Compared with the control group,there was a significant difference in richness and diversity in the HSPN group(P<0.05).At the same time,there was no significant difference in richness and diver-sity in the HSP group(P>0.05).Compared with the HSP group,the abundance,and diversity of the HSPN group were significantly increased(P<0.05).At the genus level,the proportion of Streptococcus in each group is the high-est.Compared with the control group,there was no significant correlation between the HSP group and the genus of bacteria.In contrast,the HSPN group showed a significant increase in the genera of Pseudomonas and Parabacteroi-des(P<0.05).Compared with the HSP group,the abundance of Pseudomonas and Parabacteroides in the HSPN group was significantly increased(P<0.05).(2)Compared with the control group,the HSPN group had 12 differen-tial metabolites involving nine metabolic pathways,such as phenylalanine metabolism;There was no significant dif-ference in metabolites and no metabolic pathway in the HSP group.Compared with the HSP group,the HSPN group has 15 differential metabolites involving nine metabolic pathways,such as phenylalanine metabolism.(3)In the HSPN and control groups,Pseudomonas and Parabacteroides negatively correlated with Phenylalanine metabolic pathway products.In the HSPN and HSP groups,Pseudomonas,Parabacteroides,and Phenylalanine metabolic path-way products were negatively correlated.The metabolites involved in phenylalanine metabolism in the oral cavity are 2-hydroxycinnamic acid,Phenylpyruvic acid,and N-acetyl-L-phenylalanine.Conclusion There is a significant dif-ference between HSPN and HSP children and healthy children.Streptococcus,Pseudomonas,and Parabacteroides may be one of the trigger factors of HSPN,and Phenylalanine metabolism may be one of the pathways in the patho-genesis of HSPN.Children with HSPN have a more pronounced imbalance in oral microbiota and greater differences in metabolic products than children with HSP.
7.An advanced machine learning method for simultaneous breast cancer risk prediction and risk ranking in Chinese population: A prospective cohort and modeling study
Liyuan LIU ; Yong HE ; Chunyu KAO ; Yeye FAN ; Fu YANG ; Fei WANG ; Lixiang YU ; Fei ZHOU ; Yujuan XIANG ; Shuya HUANG ; Chao ZHENG ; Han CAI ; Heling BAO ; Liwen FANG ; Linhong WANG ; Zengjing CHEN ; Zhigang YU
Chinese Medical Journal 2024;137(17):2084-2091
Background::Breast cancer (BC) risk-stratification tools for Asian women that are highly accurate and can provide improved interpretation ability are lacking. We aimed to develop risk-stratification models to predict long- and short-term BC risk among Chinese women and to simultaneously rank potential non-experimental risk factors.Methods::The Breast Cancer Cohort Study in Chinese Women, a large ongoing prospective dynamic cohort study, includes 122,058 women aged 25-70 years old from the eastern part of China. We developed multiple machine-learning risk prediction models using parametric models (penalized logistic regression, bootstrap, and ensemble learning), which were the short-term ensemble penalized logistic regression (EPLR) risk prediction model and the ensemble penalized long-term (EPLT) risk prediction model to estimate BC risk. The models were assessed based on calibration and discrimination, and following this assessment, they were externally validated in new study participants from 2017 to 2020.Results::The AUC values of the short-term EPLR risk prediction model were 0.800 for the internal validation and 0.751 for the external validation set. For the long-term EPLT risk prediction model, the area under the receiver operating characteristic curve was 0.692 and 0.760 in internal and external validations, respectively. The net reclassification improvement index of the EPLT relative to the Gail and the Han Chinese Breast Cancer Prediction Model (HCBCP) models for external validation was 0.193 and 0.233, respectively, indicating that the EPLT model has higher classification accuracy.Conclusions::We developed the EPLR and EPLT models to screen populations with a high risk of developing BC. These can serve as useful tools to aid in risk-stratified screening and BC prevention.
8.Thyroglossal Duct Papillary Carcinoma:Report of One Case and Literature Review
Guofeng DAI ; Shuya LI ; Yiqiang YIN ; Gangpu WANG
Acta Academiae Medicinae Sinicae 2024;46(5):792-796
Thyroglossal duct papillary carcinoma is extremely rare in clinical practice.This article reports a case of a thyroglossal duct cyst complicated by thyroid papillary carcinoma,while also reviewing the relevant literature.It summarizes the clinical manifestations,imaging characteristics,and pathological features of thyro-glossal duct papillary carcinoma and discusses the relationship between thyroglossal duct cysts,malignant trans-formation of thyroglossal duct cysts,and thyroid papillary carcinoma.
9.Analysis of ABO system hemolytic disease of the fetus and newborn in 1 040 cases and its influencing factors
Shuling DONG ; Xin LIU ; Ying XIE ; Shuya WANG ; Yinghui CHENLI ; Xianping LYU
Chinese Journal of Blood Transfusion 2023;36(7):605-609
【Objective】 To analyze ABO system hemolytic disease of the fetus and newborn (HDFN) and its influencing factors in Obstetrics Department of our hospital. 【Methods】 The blood samples of 1 040 neonates and their mothers in the obstetric department of our hospital were retrospectively analyzed from September 2022 to January 2023, including ABO and RhD blood group of the neonates and mothers, as well as 3 tests of HDFN, Hb, total bilirubin (TBIL) and indirect bilirubin(IBIL) of the neonates. Relevant clinical data of the neonates and mothers were collected, including maternal and neonatal age, neonatal sex, maternal pregnancy history, gestational age and delivery mode, and their influences on ABO-HDFN were analyzed. 【Results】 Among 1 040 HDFN samples, 298 were ABO incompatibility, among which 113 were HDFN positive, with a positive rate of 37.9% (113/298); the positive rate of HDFN in neonates born to mothers with type O was significantly higher than that in neonates born to mothers with type A and B (71.4% vs 8.2%, P<0.05); the positive rate of HDFN in neonates with antigen-A incompatibility was significantly higher than that in neonates with antigen-B incompatibility (48.7%vs 26.7%, P<0.05), which was the highest in neonates with O-A incompatibility [83.6% (61/73)], followed by O-B incompatibility [58.2% (39/67)]. There was no significant difference in Hb and bilirubin among the other groups except for the difference of Hb between the O-A incompatibility HDFN positive group and the HDFN negative group [(145.0±16.0) vs(153.4±13.2), P<0.05)]. The levels of Hb, TBIL and IBIL in the "direct antiglobulin test+ indirect antiglobulin test+release test+" group were significantly different from those in the HDFN negative group[(144.9±21.6) vs (153.3±13.2), P <0.05; (36.9±11.8) vs (29.6±6.1), P<0.05; (30.6±12.7) vs (23.0±6.9), P<0.05, respectively]. Logistic regression analysis showed that maternal delivery frequency, mother-neonate incompatible antigen and maternal blood type were independent risk factors for HDFN. 【Conclusion】 ABO-HDFN occurred mainly in neonates born to O-type mothers, and the positive rate was the highest in neonates with O-A incompatibility. The severity of HDFN had little relationship with the mother-neonate blood type, but had relationship with the result of 3 tests of HDFN. Maternal delivery frequency, mother-neonate incompatible antigen and maternal blood type were independent risk factors for HDFN.
10.Analysis of red blood cell alloimmunization among inpatients: 104 588 cases
Shuling DONG ; Jie SONG ; Shuya WANG ; Qiankun YANG ; Xianping LYU
Chinese Journal of Blood Transfusion 2023;36(3):245-248
【Objective】 To analyze the yield, specificity and detection time of red blood cell(RBC)alloimmunization in 104 588 inpatients. 【Methods】 The clinical information of patients who underwent at least one antibody screening in our hospital from November 2017 to December 2019 was retrospectively analyzed. The demographic characteristics, transfusion history, pregnancy history and antibody screening results of patients were collected. The RBC alloantibody yield, specificity and detection time were analyzed, and differences of transfusion units and frequency between patients with and without alloimmunization were compared. 【Results】 Eight hundred cases of alloantibodies with clinical significance were detected in blood samples of 723 patients, with a positive rate of 0.7% (723/104 588). The incidence rate of alloimmunization in females was higher than that in males (0.9% vs 0.5%, P<0.05). Rh alloantibodies accounted for 76.4%(611/800), of which 61.4%(375/611)were anti-E. Transfusion units and frequency of patients with alloimmunity were higher than those without(median: 6.0 vs 4.0, P<0.05; 4.0 vs 2.0, P<0.05, respectively). And 67.5% of RBC alloantibodies were detected within 6 months, with the median (IQR) detection time of 97.0 (22.5-247.0) days. 【Conclusion】 Routine antibody screening should be performed before transfusion in order to reduce the occurrence of adverse reactions, and Rh typing transfusion with compatible crossmatch should be performed if necessary.

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