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.Research progress on aortic root repair by modified sandwich technique for acute Stanford type A aortic dissection
Chuang LIU ; Shuya FAN ; Yangxue SUN ; Hongwei GUO
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2024;31(03):478-484
Acute Stanford type A aortic dissection has the characteristics of acute onset, severe condition and high mortality. Once making a definite diagnosis, surgical treatment is needed as soon as possible. It is difficult for cardiac surgeons to treat the acute aortic dissection involving the aortic sinus, which is an important risk factor for death. Improving the surgical treatment for the aortic sinus can be a key to improving the prognosis. In this review, we will introduce the modified sandwich technique for acute Stanford type A aortic dissection and the prognosis, and summarize the experiences of different modified sandwich techniques. However, there is still no unified standardized technique in aortic root repair, and there is a lack of large studies with long-term follow-up, so it is necessary to further improve the aortic root repair techniques.
3.Da Chengqitang Protects Intestinal Barrier in Septic Mice via Endogenous Antimicrobial Peptide mCRAMP
Yuanyuan LI ; Xuan ZHONG ; Lei LIU ; Shuya SUN ; Jiajun LING ; Rongfeng LIN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(6):20-28
ObjectiveMolecular docking and animal experiments were employed to explore the protective effect and mechanism of Da Chengqitang (DCQD) on intestinal barrier in septic mice. MethodText mining method was used to screen the active ingredients in DCQD. AutoDock Tools and Discovery Studio were used to study the interactions of active components with the core target proteins [claudin-1, tumor necrosis factor (TNF)-α, interleukin (IL)-6, endogenous antimicrobial peptide mCRAMP, Toll-like receptor 4 (TLR4), and myeloid differentiation primary response gene 88 (MyD88)] in sepsis. Fifty C57BL/6 mice were randomized into sham, model, low- and high-dose (4 g∙kg-1 and 8 g∙kg-1) DCQD, and ulinastatin groups (n=10). Before, during, and after the day of modeling surgery, each group was administrated with corresponding drugs. The mice in other groups except the model group were subjected to modeling by cecal ligation and puncture. Enzyme-linked immunosorbent assay (ELISA) was used measure the serum level of D-lactic acid to assess intestinal mucosa permeability. Hematoxylin-eosin staining was employed to observe the histopathological changes in the ileum and assess the intestinal mucosal damage and inflammatory infiltration. Western blotting was employed to determine the expression levels of tight junction proteins claudin-1 and occludin in the ileal tissue, which were indicative of the bowel barrier function. The TNF-α and IL-6 levels were measured by ELISA to assess the intestinal inflammation. The expression of mCRAMP in the ileal tissue was observed by immunohistochemistry. The mRNA levels of mCRAMP, TLR4, and MyD88 in mouse ileal tissue were determined by Real-time polymerase chain reaction, on the basis of which the mechanism of DCQD in protecting the intestinal barrier of septic mice was explored. ResultMolecular docking results showed that most of the 10 active ingredients of DCQD that were screened out by text mining could bind to sepsis targets by van der Waals force, hydrogen bonding, and other conjugated systems. The results of animal experiments showed that compared with the model group, low- or high-dose DCQD lowered the D-lactic acid level in the serum (P<0.01), alleviated damage to the ileal tissue and mucosal edema, protected the small intestine villus integrity, reduced inflammatory cell infiltration, promoted the expression of claudin-1 (P<0.01), lowered the IL-6 level (P<0.01), up-regulated the mRNA and protein levels of mCRAMP (P<0.01), and down-regulated the mRNA and protein levels of TLR4 and MyD88 (P<0.01) in the ileal tissue. In addition, high-dose DCQD lowered the TNF-α level and promoted the expression of occludin in the ileum tissue (P<0.01), and low-dose DCQD up-regulated the protein level of occludin in the ileum tissue (P<0.05). ConclusionDCQD has a protective effect on intestinal barrier in septic mice. It can reduce intestinal inflammation, repair intestinal mucosal damage, improve the tight junction protein level, and reduce intestinal mucosal permeability by up-regulating the mRNA and protein levels of mCRAMP and the down-regulating the expression of genes in the TLR4/MyD88 pathway.
4.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.
5.Analysis of the management status of emergency and life support medical equipment in township health centers in Huazhou city
Canliang FANG ; Shuya LIU ; Jun YANG ; Yi WEN ; Yuanmiao ZHOU ; Yunlian XUE ; Guihao LIU
Modern Hospital 2024;24(10):1522-1526
Objective To understand the management status of emergency and life support medical equipment in town-ship health centers in Huazhou City,providing a reference for improving emergency service levels in grassroots medical institu-tions.Methods A convenience sampling method was used to conduct a questionnaire survey of physicians,nurses,medical technicians,public health personnel,and administrative and logistical staff in 24 township health centers in Huazhou City.The survey focused on seven aspects:procurement and configuration,upgrades,operational use,maintenance,emergency handling,management systems,and quality assessment and monitoring.Results Training on emergency and life support equipment was generally adequate in Huazhou City(65.2%).Health centers in northern and central Huazhou performed better than those in the urban and southern areas regarding equipment training,emergency handling procedures for equipment failures,regular emergency drills,meeting clinical treatment needs,management system completeness,routine maintenance and inspections,and regular quality assessments.Factor analysis revealed significant issues with equipment aging and quality in urban township health cen-ters,while northern township health centers urgently needed life-sustaining and therapeutic equipment.Conclusion There are regional disparities in the management levels of emergency and life support medical equipment in township health centers in Hua-zhou City.It is recommended to enhance the overall quality of personnel in township health centers,optimize"effective training"for staff,and improve annual management plans to further elevate grassroots emergency service capabilities.
6.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.
7.Contrast-enhanced computed tomography radiomics for the preoperative prediction model of microvascular invasion in intrahepatic cholangiocarcinoma
Zheyu ZHOU ; Shuya CAO ; Chunlong ZHAO ; Qiaoyu LIU ; Xiaoliang XU ; Chaobo CHEN
International Journal of Surgery 2024;51(8):511-516
Objective:To predict the status of microvascular invasion (MVI) in intrahepatic cholangiocarcinoma (ICC) patients preoperatively based on the radiomics analysis of contrast-enhanced CT to provide imaging evidence for early identification of patients at high risk of recurrence.Methods:Clinical data of 40 ICC patients who underwent radical hepatectomy at Nanjing Drum Tower Hospital from January 2021 to May 2024 were retrospectively collected. Patients were divided into the MVI group ( n=8) and the non-MVI group ( n=32) according to the MVI status of the postoperative pathology report. Whether there were differences in each pathological index between the groups and the efficacy of radiomics analysis of contrast-enhanced CT for the preoperative prediction of MVI were analyzed. The regions of interest (ROI) were outlined on the arterial and venous phase images using the 3D Slicer software. Then, radiomics features were extracted from each ROI based on Python. Finally, the LASSO regression and glm function were used to screen radiomics features and establish a prediction model based on the R language. The established predictive model′s diagnostic efficacy, calibration, and net clinical benefit were evaluated using the receiver operating characteristic (ROC) curve, calibration curve, and decision curve analysis (DCA), respectively. Normally distributed measurement data were expressed as mean±standard deviation ( ± s) and compared using the t-test. Count data were expressed as frequency and compared using the chi-square test. Results:Patients in the MVI group had more poorly differentiated tumors and a significantly higher proportion of lymph node metastases ( P<0.05). The established radiomics prediction model included six features, 1 first-order statistical feature and 5 gray texture features. The area under the ROC curve was 0.87, the sensitivity was 75.0%, and the specificity was 90.6%. The calibration curve showed good agreement between the predicted MVI and actual MVI status, and the decision curve demonstrated that the model could provide a large net clinical benefit. Conclusion:Radiomics analysis of contrast-enhanced CT can identify the MVI status of ICC patients preoperatively and aid in clinical decision-making, providing vital evidence for individualized and precise treatment of ICC.
8.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.
9.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.
10.Research progress of psychological and biological mechanism of childhood trauma affecting social anxiety disorder
Yitong LIU ; Shuya YAN ; Jingyan SUN ; Shuming ZHONG ; Yanbin JIA
Chinese Journal of Nervous and Mental Diseases 2024;50(9):565-569
Social anxiety disorder has a significant negative impact on individuals'social interaction and normal life,and childhood trauma plays an important role in the occurrence and development of SAD.Childhood trauma affects the development of self-awareness,impairs the ability of information processing,hinders the normal development of prefrontal cortex-limbic system loop and default mode network,and causes abnormal secretion of glucocorticoid and oxytocin,which leads to individuals'inability to correctly understand social clues and reasonably regulate emotions,and thus unable to produce adaptive emotional and behavioral responses in social situations,which may lead to SAD.In conclusion,childhood trauma has a lasting adverse effect on social function from both psychological and physiological aspects.

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