[Objective] To study the molecular biological mechanism for a case of ABO blood group B subtype, and perform three-dimensional modeling of the mutant enzyme. [Methods] The ABO phenotype was identified by the tube method and microcolumn gel method; the ABO gene of the proband was detected by sequence-specific primer polymerase chain reaction (PCR-SSP), and the exon 6 and 7 of the ABO gene were sequenced and analyzed. Homologous modeling of Bw.03 glycosyltransferase (GT) was carried out by Modeller and analyzed by PyMOL2.5.0 software. [Results] The weakening B antigen was detected in the proband sample by forward typing, and anti-B antibody was detected by reverse typing. PCR-SSP detection showed B, O gene, and the sequencing results showed c.721 C>T mutation in exon 7 of the B gene, resulting in p. Arg 241 Trp. Compared with the wild type, the structure of Bw.03GT was partially changed, and the intermolecular force analysis showed that the original three hydrogen bonds at 241 position disappeared. [Conclusion] Blood group molecular biology examination is helpful for the accurate identification of ambiguous blood group. Homologous modeling more intuitively shows the key site for the weakening of Bw.03 GT activity. The intermolecular force analysis can explain the root cause of enzyme activity weakening.

Objective:To analyze the breast cancer-related parameters and the ultrasonic features of positive axillary lymph nodes,to discuss the risk factors for axillary lymphnode metastatic burden,and to provide basis for preoperative evaluation of breast cancer patients.Methods:The ultrasonic and clinicopathological data of 574 breast cancer patients with axillary lymph node metastasis confirmed by surgery and pathology were retrospectively analyzed.According to the status of axillary lymphnode metastasis,the patients were divided into low nodal burden(LNB)group(n=283)and high nodal burden(HNB)group(n=291).The affected side,tumor quadrant,distance to skin,maximum diameter,internal echogenicity,shape,margin,calcification,blood supply,posterior echo,lymphnode long diameter,lymphnode short diameter,lymphnode aspect ratio,number of suspicious metastases,intranodal blood supply,lymphnode hilum morphology,age,pathological type,histological grade,molecular subtype,and the expressions of estrogen receptor(ER),progesterone receptor(PR),Ki-67,human epidermal growth factor receptor 2(HER2),and P53 were compared between two groups.Logistic regression was used to analyze the risk factors for axillary lymph node metastatic burden in the breast cancer patients;receiver operating characteristic(ROC)curve and area under the curve(AUC)were used to evaluate the predictive value.Results:The univariate analysis results showed that there were statistically significant differences in tumor quadrant,distance to skin,molecular subtype,HER2 positive expression,lymphnode long diameter,lymph node short diameter,lymph node aspect ratio,number of suspicious metastases,and lymphnode hilum morphology between two groups(P<0.05).The multivariate Logistic regression analysis results showed that tumor located in the upper outer quadrant(OR=0.648,P=0.021),distance to skin<5 mm(OR=0.283,P=0.016),Luminal A(OR=1.564,P=0.044),lymphnode long diameter≥20 mm(OR=2.050,P<0.01),lymphnode short diameter≥8.6 mm(OR=2.430,P<0.01),lymph node aspect ratio<2(OR=1.585,P<0.01),and indistinct lymphnode hilum structure(OR=2.092,P<0.01)were the independent risk factors for axillary lymphnode metastatic burden.The ROC curve analysis results showed that compared with the ultrasonic features of positive axillary lymph nodes,the AUC of the combination of breast cancer-related parameters and ultrasonic features of positive axillary lymphnodes was larger(Z=2.72,P=0.006 5),and it had higher predictive value for axillary lymphnode metastatic burden.Conclusion:The tumor quadrant,distance to skin,molecular subtype,lymphnode long diameter,lymph node short diameter,lymphnode aspect ratio,and lymphnode hilum structure are the independent risk factors for axillary lymphnode metastatic burden,and they have certain predictive value for axillary lymphnode metastatic burden.

OBJECTIVE: To explore the molecular mechanism of a case with RhD-- phenotype. METHODS: A proband with RhD-- phenotype who attended the clinic of the First Affiliated Hospital of Zhengzhou University on January 29, 2024 was selected as the study subject. Peripheral blood samples were collected from the proband (8 mL) and her close relatives (father, mother and brother; 3 mL each) for Rh phenotyping and irregular antibodies testing with gel card and test tube methods. Direct agglutination reaction and absorption-elution test were used to detect the c antigen on the red blood cells of the proband. PCR-sequence specific primers (PCR-SSP) typing and gene sequencing were used to determine the RHCE gene of the proband and her relatives. The origin of the proband's variant was traced by pedigree analysis. Three-dimensional structural models of the wild-type RhCE*cE protein and the RhD-- phenotype protein were constructed to predict the alterations of the RhD-- phenotype protein caused by the variant. The procedures of this study were approved by the Medical Ethics Committee of the First Affiliated Hospital of Zhengzhou University (Ethics No.: 2023-KY-0870-003). RESULTS: The red blood cells of the proband did not agglutinate with anti-C, anti-c, anti-E, and anti-e. The result of the serum irregular antibody test was negative. The results of direct agglutination reaction and absorption-elution test of the proband were both negative. Her Rh blood group was identified as RhD--. The results of the Rh blood grouping of her close relatives were normal. PCR-SSP detection showed that the RHCE genotypes of the proband and her close relatives were cE/cE and Ce/cE, respectively. Gene sequencing analysis showed that the RHCE genotypes of the proband and her close relatives were RHCE*cE (c.365C>A)/RHCE*cE (c.365C>A) and RHCE*Ce/RHCE*cE (c.365C>A), respectively. Pedigree analysis revealed that the variants in the proband were inherited from her father and mother, respectively. Homology modeling of RhCE*cE protein showed that the RhD-- type peptide chain with a significantly shortened C-terminal was encoded by only 121 amino acid resides, which was 296 amino acid resides shorter compared to the wild-type RhCE*cE peptide chain encoded by 417 amino acid residues. CONCLUSION Above results revealed the molecular biological mechanism of a RhD-- phenotype. The c.365C>A variant in the RHCE gene has rendered the RHCE*cE alleles invalid, which ultimately led to the RhD-- phenotype.
Humans ; Rh-Hr Blood-Group System/chemistry* ; Female ; Phenotype ; Male ; Alleles ; Pedigree ; Base Sequence ; Molecular Sequence Data ; Adult

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.

Objective:To explore the risk factors of perioperative outcomes of lung transplantation and establish a predictive model for delayed extubation after lung transplantation.Methods:From January 1, 2020 to December 31, 2022, 104 lung transplantation recipients were retrospectively collected to identify the risk factors of early post-operative outcome.According to the timing of extubation post-lung transplantation, they were assigned into two groups of normal(77 cases)and delayed(27 cases). Baseline profiles, type of primary diagnosis, cold ischemic duration and lung transplantation approach were compared between two groups.The factors with significant difference were examined by univariate and multivariate Logistic regression.Furthermore, multivariate logistic model was visualized by a nomogram.Receiver operating characteristic(ROC)curve and decision curve analysis(DCA) were performed for evaluating the model's predictive performance and its value for clinical utilization.Results:The postoperative mortality rate was 9.6%.Delayed extubation was a strong predictor for postoperative mortality.Cold ischemic time outperformed others variates in terms of delayed extubation prediction.AUC of cold ischemic time and multivariate logistic model was 0.75(95% CI: 0.69-0.81)and 0.87(95% CI: 0.82-0.91). Conclusions:Delayed postoperative extubation is a key predictor of early post-lung transplantation mortality.The established predictive model may effectively identify high-risk patients for preventive intervention and survival improvement post-lung transplantation.

Objective:To explore the airway pathogen characteristics and examine the correlation between donor-derived pathogens and post-transplant outcomes in patients after lung transplantation (LT).Methods:Between January 1, 2015 and December 31, 2019, retrospective review was conducted for clinical and microbiological data of 88 LT recipients.Airway pathogen percentage of different microorganisms and evolution of drug-resistance were examined.Drug-resistant pathogen positive group (n=71) and negative group (n=17) were assigned according to whether or not drug-resistant pathogens were detected.Survival analysis was conducted by Log-rank with 3-year follow-ups.Between April 11, 2020 and September 5, 2020, prospective study was conducted in 14LT recipients.The potential pathogenic bacteria from donor lungs were detected by metagenomic next generation sequencing and the impact of those bacteria was examined on 1-year post-transplantation outcome in 2020.Microbial diversity and richness were shown with Shannon index.The outcome variables included heart rate, neutrophil count, lymphocyte count, immunoglobulin level and pulmonary spirometry.ANOVA and Pearson's correlation analysis were performed for elucidating the relationship between airway microbiota and post-LT outcomes.Results:From 2015 to 2019, 88 recipients were recruited and 992 strains of airway pathogens were isolated, including bacteria 796 strains and fungi 196 strains.Gram-negative bacteria (704 strains) accounted for 88.4% of all bacteria.The detection rates of Gram-positive bacteria, Klebsiella pneumonia (Kp), Acinetobacter baumannii (Ab), Stenotrophomonas maltophilia and Candida increased in 2019 than that in 2015 (8.2% vs. 5.3%, 13.6% vs. 13.2%, 33.2% vs. 17.5%, 6.5% vs. 5.3%, 26.6% vs. 20.2%). Drug resistance rate of Kp to imipenem was 68.18% in 2019 and drug resistance rate of Ab to imipenem 98.44%.The 3-year survival rate was 46.3% and 35.3% in drug-resistance positive and negative groups and the difference was insignificant ( P=0.410). Fourteen recipients were enrolled in 2020.Potential pathogenic bacteria could be detected in all donor samples.Five recipients carried the same bacteria and two died during 1-year follow-up.Nine recipients did not carry the donor-derived pathogens and two died during 1-year follow-up.The diversity of donor/recipient-derived airway microbiota (Shannon index) showed no correlation with the outcomes of 1-year follow-up by Pearson's correlation test. Conclusions:Gram-negative bacteria predominated in airway pathogens of recipients post-LT.The drug resistance rate to imipenem remained high.The donor/recipient-derived pathogen isolates showed no correlation with immediate outcomes post-LT.

Objective    To compare the clinical effects of segmentectomy and lobectomy for ≤2 cm lung adenocarcinoma with micropapillary and solid subtype negative by intraoperative frozen sections. Methods    The patients with adenocarcinoma who received segmentectomy or lobectomy in multicenter from June 2020 to March 2021 were included. They were divided into two groups according to a random number table, including a segmentectomy group (n=119, 44 males and 75 females with an average age of 56.6±8.9 years) and a lobectomy group (n=115, 43 males and 72 females with an average of 56.2±9.5 years). The clinical data of the patients were analyzed. Results    There was no significant difference in the baseline data between the two groups (P>0.05). No perioperative death was found. There was no statistical difference in the operation time (111.2±30.0 min vs. 107.3±34.3 min), blood loss (54.2±83.5 mL vs. 40.0±16.4 mL), drainage duration (2.8±0.6 d vs. 2.6±0.6 d), hospital stay time (3.9±2.3 d vs. 3.7±1.1 d) or pathology staging (P>0.05) between the two groups. The postoperative pulmonary function analysis revealed that the mean decreased values of forced vital capacity and forced expiratory volume in one second percent predicted in the segmentectomy group were significantly better than those in the lobectomy group (0.2±0.3 L vs. 0.4±0.3 L, P=0.005; 0.3%±8.1% vs. 2.9%±7.4%, P=0.041). Conclusion    Segmentectomy is effective in protecting lungs function, which is expected to improve life quality of patients.

OBJECTIVE: To explore the molecular basis for an A subtype of the ABO blood group. METHODS: The forward and reverse typing of the ABO blood group were identified by gel card and test tube methods. The ABO gene of the patient was detected by PCR-sequence specific primer (PCR-SSP). Exons 1 to 7 of the ABO gene was amplified by PCR and sequenced. The ABO gene was also subjected to subclone sequencing for haplotype analysis. RESULTS: The patient's red cells showed weak agglutination with anti-A but non-agglutination with anti-B. The patient's serum showed 1+ agglutination with A cells and 4+ agglutination with B cells. Based on above serological characteristics, the patient was defined as Aw subtype of the ABO blood group. Sequencing analysis showed that the patient was heterozygous for c.106G>T, c.188G>A, c.189C>T, c.220C>T, c.297A>G, c.467C>T, c.543G>C, c.646T>A, c.681G>A, c.771C>T, c.829G>A, in addition with a c.261G deletion. Combined with the result of subclone sequencing, the ABO genotype of the patient was determined as ABO*AW.33. new/O.01.02, which harbored c.467C>T and c.543G>C variants compared with ABO*A1.01 and c.543G>C variant compared with ABO*A1.02. The novel allele has been submitted to GenBank with an accession number of MK302122. CONCLUSION A novel allele of Aw33 subtype has been identified with its GTA transferase gene harboring c.467C>T and c.543G>C variants compared with A1.01.
ABO Blood-Group System ; genetics ; Alleles ; Exons ; genetics ; Genotype ; Humans ; Phenotype

Objective    To analyze whether hypernatremia within 48 hours after cardiac surgery will increase the incidence of delirium which developed 48 hours later after surgery (late-onset delirium). Methods    We conducted a retrospective analysis of 3 365 patients, including 1 918 males and 1 447 females, aged 18-94 ( 60.53±11.50) years, who were admitted to the Department of Cardiothoracic and Vascular Surgery of Nanjing First Hospital and underwent cardiac surgery from May 2016 to May 2019. Results    A total of 155 patients developed late-onset delirium, accounting for 4.61%. The incidence of late-onset delirium in patients with hypernatremia was 9.77%, the incidence of late onset delirium in patients without hypernatremia was 3.45%, and the difference was statistically different (P<0.001). The odds ratio (OR) of hypernatremia was 3.028 (95% confidence interval: 2.155-4.224, P<0.001). The OR adjusted for other risk factors including elderly patients, previous history of cerebrovascular disease, operation time, cardiopulmonary bypass time, lactate, hemoglobin≥100 g/L, prolonged mechanical ventilation, left ventricular systolic function, use of epinephrine, use of norepinephrine was 1.524 (95% confidence interval: 1.031-2.231, P=0.032). Conclusion    Hypernatremia within 48 hours after cardiac surgery may increase the risk of delirium in later stages.

Background and objective As computed tomography (CT) screening for lung cancer becomes more common in China, so too does detection of pulmonary ground-glass nodules (GGNs). Although anumber of national or international guidelines about pulmonary GGNs have been published,most of these guidelines are produced by respiratory, oncology or radiology physicians, who might not fully understand the progress of modern minimal invasive thoracic surgery, and these current guidelines may overlook or underestimate the value of thoracic surgery in the management of pulmonary GGNs. In addition, the management for pre-invasive adenocarcinoma is still controversial. Based onthe available literature and experience from Shanghai Pulmonary Hospital, we composed this consensus about diagnosis and treatment of pulmonary GGNs. For lesions which are considered as adenocarcinoma in situ, chest thin layer CT scan follow-up is recommended and resection can only be adopt in some specific cases and excision should not exceed single segment resection. For lesions which are considered as minimal invasive adenocarcinoma, limited pulmonary resection or lobectomy is recommended. For lesions which are considered as early stage invasive adenocarcinoma, pulmonary resection is recommend and optimal surgical methods depend on whether ground glass component exist, location, volume and number of the lesions and physical status of patients. Principle of management of multiple pulmonary nodules is that primary lesions should be handled with priority, with secondary lesions taking into account.
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Adenocarcinoma ; diagnosis ; diagnostic imaging ; surgery ; Adenocarcinoma of Lung ; China ; Consensus ; Hospitals ; Humans ; Lung Neoplasms ; diagnosis ; diagnostic imaging ; surgery ; Physicians ; psychology ; Positron Emission Tomography Computed Tomography ; Practice Guidelines as Topic ; Retrospective Studies ; Solitary Pulmonary Nodule ; diagnosis ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed