Objective: To reveal the molecular biological mechanism of a rare Dc-variant individual using PacBio third-generation sequencing technology. Methods: ABO and Rh blood type identification, DAT, unexpected antibody screening and D antigen enhancement test were conducted by serological testing. The absorption-elution test was used to detect the e antigen. RHCE gene typing was performed by PCR-SSP, and the 1-10 exons of RHCE were sequenced by Sanger sequencing. The full-length sequences of RHCE, RHD and RHAG were detected by PacBio third-generation sequencing technology. Results: Serological findings: Blood type O, Dc-phenotype, DAT negative, unexpected antibody screening negative; enhanced D antigen expression; no detection of e antigen in the absorption-elution test. PCR-SSP genotyping indicated the presence of only the RHCE c allele. Sanger sequencing results: Exons 5-9 of RHCE were deleted, exon 1 had a heterozygous mutation at c. 48G/C, and exon 2 had five heterozygous mutations at c. 150C/T, c. 178C/A, c. 201A/G, c. 203A/G and c. 307C/T. Third-generation sequencing results: RHCE genotype was RHCE 02N. 08/RHCE-D(5-9)-CE; RHD genotype was RHD 01/RHD 01; RHAG genotype was RHAG 01/RHAG 01 (c. 808G>A and c. 861G>A). Conclusion: This Dc-individual carries the allele RHCE 02N. 08 and the novel allele RHCE-D(5-9)-CE. The findings of this study provide data support and a theoretical basis for elucidating the molecular mechanisms underlying RhCE deficiency phenotypes.

[Abstract] [Objective] To evaluate the immunogenicity of red blood cell blood group antigens in the population of Xi'an. [Methods] Data on blood group antigens of voluntary blood donors from the Shaanxi Province Blood Center and unexpected antibody detection results from clinically submitted cases between January 2019 and May 2024 were analyzed. The Giblett blood group antigen immunogenicity calculation formula was used to calculate the immunogenicity of blood group antigens based on the frequency of unexpected antibodies and the probability of antigen-negative patients receiving antigen-positive red blood cells. The relative immunogenicity of each blood group antigen was obtained by multiplying the immunogenicity of the K antigen (0.095). [Results] A total of 30 921 individuals were included for red blood cell blood group antigen analysis, with 511 cases of unexpected antibody identification. The ranking of red blood cell blood group antigen immunogenicity for the overall population was: Wr^a>E>Di^b>Fy^a>K>C>e>c>Di^a>Jk^a>M>Le^a>Jk^b>Le^b>Fy^b>S, while for males, it was: Di^b>Wr^a>E>K>Fy^a>C>e>c>M>Di^a>Jk^a>Fy^b>Le^a>Le^b>Jk^b>S. [Conclusion] Based on the immunogenicity ranking from strong to weak of red blood cell antigens in the population of Xi'an, this study provides theoretical support for the expansion and matching of antigens, and technical support for achieving precise red blood cell transfusions to improve transfusion efficacy and safety.

Background::Triple-negative breast cancer (TNBC) is a type of highly invasive breast cancer with a poor prognosis. According to new research, long noncoding RNAs (lncRNAs) play a significant role in the progression of cancer. Although the role of lncRNAs in breast cancer has been well reported, few studies have focused on TNBC. This study aimed to explore the biological function and clinical significance of forkhead box C1 promoter upstream transcript (FOXCUT) in triple-negative breast cancer.Methods::Based on a bioinformatic analysis of the cancer genome atlas (TCGA) database, we detected that the lncRNA FOXCUT was overexpressed in TNBC tissues, which was further validated in an external cohort of tissues from the General Surgery Department of the First Affiliated Hospital of Nanjing Medical University. The functions of FOXCUT in proliferation, migration, and invasion were detected in vitro or in vivo. Luciferase assays and RNA immunoprecipitation (RIP) were performed to reveal that FOXCUT acted as a competitive endogenous RNA (ceRNA) for the microRNA miR-24-3p and consequently inhibited the degradation of p38. Results::lncRNA FOXCUT was markedly highly expressed in breast cancer, which was associated with poor prognosis in some cases. Knockdown of FOXCUT significantly inhibited cancer growth and metastasis in vitro or in vivo. Mechanistically, FOXCUT competitively bounded to miR-24-3p to prevent the degradation of p38, which might act as an oncogene in breast cancer. Conclusion::Collectively, this research revealed a novel FOXCUT/miR-24-3p/p38 axis that affected breast cancer progression and suggested that the lncRNA FOXCUT could be a diagnostic marker and therapeutic target for breast cancer.

Objective To investigate the clinical features in children with invasive pneumococcal disease (IPD) and early warning indicators of severe case.Methods The case data of 101 children inpatients with IPD isolated Streptococcus pneumoniae (Sp) at least once blood culture in this hospital from January 2007 to De-cember 2021 were collected retrospectively.The children patients were divided into the pediatric intensive care unit (PICU) group (35 cases) and general ward group (66 cases) according to whether or not entering PICU during hospitalization.The general clinical data,infection types and results of drug sensitivity test and bacteri-al strain serotype identification were compared between the two groups and analyzed.Results Compared with the general ward group,the male proportion,proportions of complicating nervous system disease and iron-defi-ciency anemia,winter incidence rate,disturbance of consciousness,convulsion,dyspnea,heart rate increase,oc-currence rate of vomiting and diarrhea,proportion of peripheral blood neutrophilic granulocytes,C reactive protein (CRP),lactic dehydrogenase (LDH) level,Hb<90 g/L,LDH>700 U/L,creatine kinase isoenzyme-MB (CK-MB) abnomality,proportions of bacterial meningitis,sepsis and severe pneumonia in the PICU group were higher,the proportion of 3-97 percentile of body weight,bronchopneumonia and no-focus blood stream infection were lower,WBC,Hb and albumin level were lower,and the differences were statistically significant (P<0.05).Eleven kinds of serotypes were identified in this study,ranking the top five in order were 6B,14,19F,23F and 19A.Pneumococcal 13-valent conjugate vaccine (PCV13) could cover 96.9％ of the IPD sero-types.The multivariate logistic regression analysis showed that consciousness disturbance,convulsion,per-centage of neutrophils increase,Hb<90 g/L,LDH>700 U/L and CK-MB abnormality indicated the condition of IPD children patients was critical,which was the early-warning indicator of entering PICU (P<0.05).Con-clusion Severe IPD lacks the early specific early-warning indicator,and the cover rate of PCV13 is high.

Objective This paper aims to construct a system integrating mixed reality technology with artificial algo-rithm and to evaluate its effectiveness in vascular localization during anterolateral thigh perforator flap surgery to provide new insights for clinical practice.Methods Twenty patients undergoing anterolateral thigh perforator flap repair were selected.After attaching positioning devices on the lower limb,CT angiography(CTA)scans were performed.The 2D data obtained were converted into a 3D model of the positioning device and vessels.Mixed reality technology was uti-lized to achieve 3D visualization of perforator vessels.An artificial algorithm was developed in HoloLens 2 to match the positioning device automatically with its 3D model intraoperatively to overlap the perforator vessels with their 3D mod-els.The number of perforator vessels identified within the flap harvesting area and the actual number detected during sur-gery were recorded to calculate the accuracy rate of vessel identification based on CTA data reconstruction.The distance between the perforator vessel exit points located by the system and the actual exit points was measured,and the error val-ues were calculated.The surgical time required for the system to harvest the anterolateral thigh perforator flap was docu-mented and compared with the surgical time required by conventional methods.The clinical applicability of the system was discussed.Results The CTA data reconstruction identified 30 perforator vessels,while the actual number found during surgery was 32,resulting in an identification accuracy rate of 93.75％.The average distance between the perfora-tor vessel exit points located by the system and the actual exit points was(1.65±0.52)mm.The average surgical time for flap harvesting with the assistance of the system was(43.45±4.6)min compared with(57.6±7.9)min required by conven-tional methods.All perforator flaps survived the procedure.One case of flap infection occurred seven days postoperative-ly,and one case of partial flap necrosis was treated with symptomatic therapy,resulting in delayed healing.Conclusion The system constructed in this paper can achieve 3D visualization of perforator vessels through mixed reality technology and improve the accuracy of perforator vessel localization using artificial algorithms,hence demonstrating potential ap-plication in anterolateral thigh perforator flap harvesting surgeries.

Objective:To explore the short-term clinical efficacy, safety and patients' prognostic influencing factors of CyberKnife for the treatment of brain metastases in non-small cell lung cancer (NSCLC).Methods:A retrospective case series study was conducted. The clinical data of 58 NSCLC patients who received CyberKnife treatment for brain metastases at Jilin Cancer Hospital from July 2020 to January 2022 were retrospectively analyzed. At 3 months after CyberKnife treatment for brain metastases, and the efficacy of radiotherapy was evaluated on the basis of changes of brain metastases detected by contrast-enhanced magnetic resonance imaging (MRI) of the head. Overall survival (OS) and local recurrence-free survival (LRRFS) were analyzed in 58 patients by using the Kaplan-Meier method; the efficacy of cumulative brain metastasis volume for determining the survival of CyberKnife-treated NSCLC patients with brain metastases was analyzed by using the receiver operating characteristic (ROC) curve with the survival status of patients during the follow-up period as the gold standard, and the optimal cut-off value of cumulative brain metastasis volume was obtained; the clinical factors affecting OS and LRRFS of CyberKnife-treated NSCLC patients with brain metastases were analyzed by univariate and multivariate Cox proportional hazards models, and the adverse reactions associated with CyberKnife treatment were evaluated.Results:Among the 58 patients, 26 (44.8%) were male and 32 (55.2%) were female, with a median age [ M ( Q1, Q3)] of 64 years old (56 years old, 70 years old); there were 1-7 brain metastatic lesions in each patient, and there were 98 brain metastatic lesions in the 58 patients. There were 2 deaths (3.4%) within 3 months after CyberKnife treatment. At 3 months after treatment, there were 3 cases (5.4%) in complete remission, 36 cases (64.3%) in partial remission, 13 cases (23.2%) in stable disease, and 4 cases (7.1%) in disease progression in the remaining 56 patients. ROC curve analysis showed that the area under the curve for determining the survival of CyberKnife-treated NSCLC patients with brain metastases based on the cumulative brain metastasis volume was 0.593 (95% CI: 0.423-0.763), and the optimal cut-off value of cumulative brain metastasis volume was 15 cm 3. Median follow-up time was 12.6 months (7.5 months, 17.9 months). The 6- and 12-month OS rates were 91.3% and 79.5%, respectively, and the 6- and 12-month LRRFS rates were 93.0% and 89.2%, respectively. Multivariate Cox regression analysis showed that the Karnofsky functional status score (>70 points vs. ≤70 points, HR= 0.103, 95% CI: 0.019-0.545, P = 0.007), control of extracranial tumor (controlled vs. uncontrolled, HR = 0.145, 95% CI: 0.049-0.429, P < 0.001), cumulative brain metastasis volume (≤15 cm 3vs. >15 cm 3, HR = 0.105, 95% CI: 0.028-0.399, P = 0.001) were independent influencing factors for poor OS, and the control of extracranial tumor (controlled vs. uncontrolled, HR = 0.062, 95% CI: 0.006-0.616, P = 0.018), cumulative brain metastasis volume (≤15 cm 3vs. >15 cm 3, HR = 0.440, 95% CI: 0.007-0.292, P = 0.001), and target area total bioequivalent dose (BED) (≤60 Gy vs. >60 Gy, HR = 5.299, 95% CI: 1.020-27.530, P = 0.047) were independent influencing factors for poor LRRFS. Only grade 1-2 headache [53.5% (31/58)], nausea and vomiting [36.2% (21/58)] and other adverse reactions occurred after treatment, and no ≥grade 3 adverse reactions occurred. Conclusions:CyberKnife treatment for NSCLC brain metastases has high local control rate and short-term survival rate with mild adverse effects. Karnofsky functional status score, control of extracranial tumor and cumulative brain metastasis volume may affect OS of CyberKnife-treated NSCLC patients with brain metastases, and the control of extracranial tumor, cumulative brain metastasis volume and total BED may affect local recurrence.

Radiotherapy is widely used in the management of advanced colorectal cancer (CRC). However, the clinical efficacy is limited by the safe irradiated dose. Sensitizing tumor cells to radiotherapy via interrupting DNA repair is a promising approach to conquering the limitation. The BRCA1-BARD1 complex has been demonstrated to play a critical role in homologous recombination (HR) DSB repair, and its functions may be affected by HERC2 or BAP1. Accumulated evidence illustrates that the ubiquitination-deubiquitination balance is involved in these processes; however, the precise mechanism for the cross-talk among these proteins in HR repair following radiation hasn't been defined. Through activity-based profiling, we identified PT33 as an active entity for HR repair suppression. Subsequently, we revealed that BAP1 serves as a novel molecular target of PT33 via a CRISPR-based deubiquitinase screen. Mechanistically, pharmacological covalent inhibition of BAP1 with PT33 recruits HERC2 to compete with BARD1 for BRCA1 interaction, interrupting HR repair. Consequently, PT33 treatment can substantially enhance the sensitivity of CRC cells to radiotherapy in vitro and in vivo. Overall, these findings provide a mechanistic basis for PT33-induced HR suppression and may guide an effective strategy to improve therapeutic gain.

Objective:To evaluate the relationship between the preoperative neutrophil/lymphocyte ratio(NLR) and postoperative delirium (POD)in elderly patients.Methods:Nine hundred and thirty-seven patients, undergoing elective knee or hip arthroplasty under combined spinal and epidural anesthesia, in whom Mini-Mental State Examination was completed at 1 day before operation, with Mini-Mental State Examination score≥24, were selected. Elbow venous blood samples were collected before surgery, neutrophils and lymphocytes were counted, and the ratio of neutrophils to lymphocytes was calculated.Cerebrospinal fluid (CSF) 2 ml was extracted after successful spinal-epidural puncture for measurement of preoperative amyloid beta40 (Aβ40), amyloid beta42 (Aβ42), total Tau (T-tau), and phosphorylated Tau (P-tau) by enzyme-linked immunosorbent assay. POD was assessed by Confusion Assessment Method, and the severity of POD was assessed by the Memorial Delirium Assessment Scale.The logistic regression equation was used to identify the risk factors for POD, and the mediating effect of CSF biomarkers was analyzed. Sensitivity analysis was used to test the stability of the results. The receiver operating characteristic curve was introduced, and the area under the curve was calculated to evaluate the accuracy of preoperative NLR in predicting POD.Results:A total of 853 patients were finally enrolled in this study, and 17.4% patients developed POD. Logistic regression analysis showed that the increased levels of NLR ( OR 1.141, 95% confidence interval [ CI] 1.033-1.260, P=0.010), P-tau in CSF ( OR 1.093, 95% CI 1.076-1.110, P<0.001) and T-tau in CSF( OR 1.003, 95% CI 1.001-1.005, P<0.001) were risk factors for POD, while the increased level of Aβ42 in CSF( OR 0.998, 95% CI 0.997-1.000, P=0.028) was a protective factor for POD after adjusting for multiple confounding factors. Analysis of mediating effect: T-tau and P-tau in CSF were the mediating factors in the relationship between NLR and POD with the mediating effects of 0.011 9 and 0.020 0 respectively, and the proportion of mediating effect was 46.1% and 53.1% respectively.The receiver operating characteristic curve showed that the area under the curve of NLR and combination of NLR and CSF biomarkers in predicting POD was 0.711 and 0.939 respectively. Conclusions:Increased preoperative NLR level is a risk factor for POD, and combination of NLR and CSF biomarkers shows a higher accuracy in predicting POD. T-tau and P-tau in CSF serve as the key mediators in the relationship between NLR and POD.

【Objective】 To evaluate the application of monoclonal typing reagents and human anti-A/B antibodies for absorption-elution test in ABO grouping. 【Methods】 The specificity of monoclonal typing reagents and human anti-A/B antibodies with standard A, B, O and AB phenotypes at 4 ℃, room temperature, and 37 ℃ were compared. Affinity was evaluated by the titer, agglutination time and agglutination intensity of the reaction with A1/B cells. 29 samples with ABO discrepancy were tested to evaluate the ability of monoclonal typing reagents and human anti-A/B antibodies to detect weak antigens in absorption-elution test. 【Results】 The specificity and affinity of human anti-A/B antibodies are low, and monoclonal typing reagents have cross reactivity. Human anti-A/B antibodies can detect most weak antigens in absorption-elution test with no cross reactivity. 【Conclusion】 In ABO grouping, the human anti A/B antibody binding absorption-elution test can serve as a supplement method for identifying ABO weak antigens. Accurate results can be obtained with reasonable reagents and corresponding methodology in serological tests,thus ensuring the safety of blood transfusion.

【Objective】 To establish a rare blood group information supply platform in Shaanxi Province. 【Methods】 The rare blood group information supply platform consists of sample registration, result registration, donor files and inventory blood. The blood donation codes of voluntary blood donors were recorded for blood typing, and the antigen identification results of each blood group system were registered, all stored in the rare blood type information supply platform. When receiving an application for unusual or rare blood type missing multiple conventional antigens or a certain high-frequency antigen, the corresponding antigen negative blood donors and their blood status (in stock or not) were queried from the donor profile module of the platform, and the inventory of blood of rare blood type was monitored dynamically. 【Results】 The results showed that 5.060% (273/5 398) of rare Rh phenotype donors, 1.540‰ (51/33 010) of donors lacking multiple regular antigens, and 13 O-type donors lacking high-frequency antigens were recorded in the rare blood type information supply platform. Among them, 0.019‰ (3/158 484) of Jk(a-b-) phenotype, 0.436‰ (2/4 586) of Di(a+b-) phenotype, and 4.030‰ (8/1 983) of Fy (a-b+) phenotype were stored in the blood bank for rare blood type. 【Conclusion】 The establishment of rare blood group information supply platform can meet the urgent demand for blood of rare blood types in clinical practice and ensure the safety of blood transfusion.