OBJECTIVE: To report the blood group antigen and antibody specificity identification methods for a patient with high-frequency antibodies, and the process of finding and providing compatible blood for the patient. METHODS: A patient sent from the Blood Transfusion Department of Shanxi Provincial People's Hospital to Blood Transfusion Technology Research Laboratory of Taiyuan Blood Center in November 2022 was selected for the study. Classical serological methods were used to determine the patient's blood type, screen for unexpected antibodies, identify antibodies, and perform crossmatching. High-frequency antibody identification was carried out using red blood cells treated with various enzymes. Blood group genotyping was conducted using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF) and Sanger sequencing. Multiple strategies were employed to address the patient's blood source problem. The study was approved by the Medical Ethics Committee of Taiyuan Blood Center [Ethics No. 2024 Ethics Review No.(2)]. RESULTS: The patient's blood type was B, RhD positive. Initial screening of the patient's serum with multiple screening cells and antibody identification cells in saline medium was negative, but positive in antiglobulin medium. The patient's serum showed varying reaction intensities with red blood cells treated with different enzymes. MALDI-TOF mass spectrometry and Sanger sequencing revealed a homozygous nonsense variant c.376C>T (p.Gln126Ter) in the ABCG2 gene, resulting in the Jr(a-) phenotype. During family donor selection, the patient's son was found to have a heterozygous variant c.376C>T (p.Gln126Ter), and another heterozygous variant c.421C>A (p.Gln141Lys), which predicted a Jr(a+w) phenotype. Crossmatch tests confirmed the compatibility of blood from the patient's son, which was used to address the urgent blood requirement. Later, rare blood from a Jr(a-) donor from the Guangzhou Blood Center was used for the patient's ongoing treatment, saving the patient's life. CONCLUSION Combining classic serological testing with blood group gene typing techniques successfully identified the rare Jr(a-) blood type and high-frequency anti-Jra antibodies. Enzyme-treated red blood cell identification methods confirmed the presence of anti-Jra antibodies. By searching within the family and seeking help from other blood centers, compatible blood was found. This approach may provide insights for resolving similar complex blood matching problems in the future.
Humans ; Blood Grouping and Crossmatching/methods* ; Blood Group Antigens/immunology* ; Blood Transfusion ; Male ; Isoantibodies/blood* ; Female ; Genotype

Objective:To report the blood group antigen and antibody specificity identification methods for a patient with high-frequency antibodies, and the process of finding and providing compatible blood for the patient.Methods:A patient sent from the Blood Transfusion Department of Shanxi Provincial People′s Hospital to Taiyuan Blood Center in November 2022 was selected for the study. Classical serological methods were used to determine the patient′s blood type, screen for unexpected antibodies, identify antibodies, and perform crossmatching. High-frequency antibody identification was carried out using red blood cells treated with various enzymes. Blood group genotyping was conducted using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF) and Sanger sequencing. Multiple strategies were employed to address the patient′s blood source problem. The study was approved by the Medical Ethics Committee of Taiyuan Blood Center [Ethics No. 2024 Ethics Review No.(2)].Results:①The patient′s blood type was B, RhD positive. Initial screening of the patient′s serum with multiple screening cells and antibody identification cells in saline medium was negative, but positive in antiglobulin medium. The patient′s serum showed varying reaction intensities with red blood cells treated with different enzymes. ②MALDI-TOF mass spectrometry and Sanger sequencing revealed a homozygous nonsense variant c. 376C>T (p.Gln126Ter) in the ABCG2 gene, resulting in the Jr(a-) phenotype. During family donor selection, the patient′s son was found to have a heterozygous variant c. 376C>T (p.Gln126Ter), and another heterozygous variant c. 421C>A (p.Gln141Lys), which predicted a Jr(a+ w) phenotype. ③Crossmatch tests confirmed the compatibility of blood from the patient′s son, which was used to address the urgent blood requirement. Later, rare blood from a Jr(a-) donor from the Guangzhou Blood Center was used for the patient′s ongoing treatment, saving the patient′s life. Conclusion:Combining classic serological testing with blood group gene typing techniques successfully identified the rare Jr(a-) blood type and high-frequency anti-Jra antibodies. Enzyme-treated red blood cell identification methods confirmed the presence of anti-Jra antibodies. By searching within the family and seeking help from other blood centers, compatible blood was found. This approach may provide insights for resolving similar complex blood matching problems in the future.

Objective:To report the blood group antigen and antibody specificity identification methods for a patient with high-frequency antibodies, and the process of finding and providing compatible blood for the patient.Methods:A patient sent from the Blood Transfusion Department of Shanxi Provincial People′s Hospital to Taiyuan Blood Center in November 2022 was selected for the study. Classical serological methods were used to determine the patient′s blood type, screen for unexpected antibodies, identify antibodies, and perform crossmatching. High-frequency antibody identification was carried out using red blood cells treated with various enzymes. Blood group genotyping was conducted using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF) and Sanger sequencing. Multiple strategies were employed to address the patient′s blood source problem. The study was approved by the Medical Ethics Committee of Taiyuan Blood Center [Ethics No. 2024 Ethics Review No.(2)].Results:①The patient′s blood type was B, RhD positive. Initial screening of the patient′s serum with multiple screening cells and antibody identification cells in saline medium was negative, but positive in antiglobulin medium. The patient′s serum showed varying reaction intensities with red blood cells treated with different enzymes. ②MALDI-TOF mass spectrometry and Sanger sequencing revealed a homozygous nonsense variant c. 376C>T (p.Gln126Ter) in the ABCG2 gene, resulting in the Jr(a-) phenotype. During family donor selection, the patient′s son was found to have a heterozygous variant c. 376C>T (p.Gln126Ter), and another heterozygous variant c. 421C>A (p.Gln141Lys), which predicted a Jr(a+ w) phenotype. ③Crossmatch tests confirmed the compatibility of blood from the patient′s son, which was used to address the urgent blood requirement. Later, rare blood from a Jr(a-) donor from the Guangzhou Blood Center was used for the patient′s ongoing treatment, saving the patient′s life. Conclusion:Combining classic serological testing with blood group gene typing techniques successfully identified the rare Jr(a-) blood type and high-frequency anti-Jra antibodies. Enzyme-treated red blood cell identification methods confirmed the presence of anti-Jra antibodies. By searching within the family and seeking help from other blood centers, compatible blood was found. This approach may provide insights for resolving similar complex blood matching problems in the future.

Objective:To investigate the clinical efficacy of the bone-forming channel technique combined with double plates in the treatment of postoperative aseptic nonunion of the humeral shaft. Methods:A retrospective study was conducted to analyze the data of 39 patients with postoperative aseptic nonunion of the humeral shaft who had been treated by bone-forming channel technique combined with double plates from March 2018 to December 2022. There were 20 males and 19 females, with an age of (46.5±1.6) years. The bone nonunion was atrophic in 18 cases and hypertrophic in 21 cases. The operation time, intraoperative bleeding, fracture healing time and complications were recorded. The shoulder and elbow functions were evaluated before operation and at the final follow-up using the Disabilities of the Arm, Shoulder and Hand (DASH) score, Neer's shoulder function score, and Mayo's elbow performance score (MEPS). Results:All the 39 patients were followed up for an average duration of (13.8±3.5) months. The operation time was (84.3±7.3) minutes, the intraoperative bleeding (127.5±4.5) mL, the fracture healing time (24.4±1.3) weeks, and the fracture healing rate 100% (39/39). None of the patients had such postoperative complications as infection, neurovascular injury, or re-rupture of internal fixation. At the final follow-up, the patients' DASH score [(7.4±1.2) points], Neer's shoulder function score [(93.4±5.2) points], and MEPS [(94.6±3.3) points] were significantly improved compared respectively with the preoperative values [(26.6±8.4) points, (74.3±6.5) points, and (73.8±5.3) points] ( P<0.05). At the final follow-up, the patients' forward flexion and upward mobility was 168.5°±3.4°, abduction and upward mobility was 167.5°±5.2°, and horizontal forward flexion was 129.5°±11.8°. Conclusion:In the treatment of postoperative aseptic nonunion of the humeral shaft, since the bone-forming channel technique combined with double plates can achieve satisfactory clinical outcomes, it is worthy of clinical application.

Objective:To investigate the clinical characteristics and significance of supraspinatus fragments in proximal humerus fractures.Methods:A total of 210 patients with proximal humerus fractures who underwent surgical treatment in the Department of Orthopaedic and Trauma Surgery, Affiliated Hospital of Yunnan University from July 2016 to December 2020 were retrospectively analyzed. There were 91 males and 119 females, aged 52.4±11.3 years (range, 18-87 years). Preoperatively, the shoulder joint X-ray, CT and 3D reconstruction, and MRI were evaluated for the presence of a small fracture mass on the lateral side of the interarticular sulcus and an "M" shaped fracture mass at the interarticular sulcus. Intraoperative observation, management, postoperative functional exercise and follow-up were performed by the same group of doctors. Postoperative outpatient review was performed at 2, 4, 6, 12, and 24 weeks, shoulder joint X-ray and CT were taken at 2, 6, and 24 weeks to observe the presence of subacromial displaced bone mass. Constant-Murley scale was used to assess shoulder joint function.Results:All 210 patients completed the surgery successfully, and none of them had vascular, nerve or ligament injuries during the operation. According to Neer's classification: 59 cases of two-part humeral greater tuberosity fractures, 36 cases of two-part humeral surgical neck fractures, 50 cases of three-part humeral greater tuberosity fractures, and 65 cases of four-part proximal humeral fractures. 61.0% (128/210) of the patients showed the presence of a supraspinatus fragment on preoperative imaging, including 42 two-part humeral greater tuberosity fractures, 1 two-part humeral surgical neck fracture, and 31 three-part humeral greater tuberosity fractures with a small lateral fracture mass over the intertubular groove; 54 four-part proximal humeral fractures had an "M" shaped fracture mass containing the intertubular groove. All patients were followed up for 11.8±2.4 months (range, 6-20 months). Postoperative X-ray showed that all fractures healed, and the healing time was 3.4±0.3 months (range, 3-5 months). The Constant-Murley scale of the shoulder joint was 86.3±11.5 (range, 61-100).Conclusion:Supraspinatus fragments are more common in proximal humerus fractures. Proper recognition and management of supraspinatus fragments can help understand the mechanism of proximal humeral fracture occurrence, the relationship between fracture displacement and rotator cuff injury, reduce the incidence of postoperative complications, and improve the shoulder joint function.

【Objective】 To analyze the influence of β-lactam antibiotics on RBC aging and clearance by detecting various indicators of aging and clearance on RBCs, as well as the differences in phagocytosis for erythrocytes before and after drugs treated in vitro. 【Methods】 RBCs were treated by β-lactam antibiotics, including Penicillin, Cefepime, Cefoperazone and Ceftazidime, and the changing of phosphatidylserine (PS) and clearance related CD markers, including CD35, CD47, CD55 and CD59 on the surface of the RBCs, were detected by flow cytometry at 0h and 24h after drugs treatment. The proportion of acanthocytes by microscope also at 0h and 24h after drugs treatment was calculated. The phagocytosis of drug-treated RBC was detected by monocyte monolayer assay (MMA). Untreated RBCs were incubated in PBS by the same condition as a negative control.The influence of β-lactam antibiotics on RBC aging and clearance by all the results above was studied. 【Results】 Compare to the untreated RBCs, the drug treated RBCs showed a higher PS level on the cell surface. The results showed by percentage as following(0 h vs 24 h): Penicillin 9.42% vs 93.30%, Cefepime 3.88% vs 57.27%, Cefoperazone 4.71% vs 75.75% and Ceftazidime 3.05% vs 43.19%. The acanthocytes ratio was as following(0 h vs 24 h): Penicillin 7.33% vs 86%, Cefepime 2.67% vs 52.67%, Cefoperazone 3.33% vs 67.67% and Ceftazidime 3.33% vs 90.67%. On the opposite, the clearance related CD markers, showed an obviously lower level after drugs treated(0 h vs 24 h): CD35: Penicillin 7.36% vs 11.87%, Cefepime 0.14% vs 28.51%, Cefoperazone 11.85% vs 21.55% and Ceftazidime 7.63% vs 8.73%; CD47: Penicillin 1.22% vs 9.13%, Cefepime 1.80% vs 0.86%, Cefoperazone 0.08% vs 6.85% and Ceftazidime 1.54% vs 5.50%; CD55: Penicillin 14.46% vs 44.31%, Cefepime 17.27% vs 38.41%, Cefoperazone 19.28% vs 33.28% and Ceftazidime 14.62% vs 34.13%; CD59: Penicillin 4.71% vs 20.56%, Cefepime 4.03% vs 7.60%, Cefoperazone 5.91% vs 22.38% and Ceftazidime 5.93% vs 30.89%. Drug-treated RBCs attached more to monocytes than untreated RBCs. 【Conclusion】 The β-lactam antibiotics could induce the changing of PS and the clearance of related CD markers on surface of RBCs. They also could lead acanthocytes and make the RBCs more susceptible to phagocytosis by monocytes. The β-lactam antibiotics could promote the RBCs aging and clearance, which might deteriorate the DIIHA.

【Objective】 To establish human hybridoma cell lines, secreting monoclonal antibody against antigens of Rh blood system, from a donor with rare D--phenotype. 【Methods】 Peripheral blood B lymphocytes of an O type female donor, lacking C/c/E/e antigens on her erythrocyte, were transformed with Epstein-Barr virus (EBVs). EBVs were harvested from the cultural supernatant of B95-8 cells. The transformed lymphoblastoid cell line (LCL) secreting antibodies to C antigens were picked up and then hybridized with the myeloma SHM-D33 using electric fusion technique. Hybridoma cells were selected by HATD-Ouabain(HOTD)(Hypoxantine, Aminopterin, Thymidine, 2-Deoxycytide, and Ouabain)culture medium, microplate antibody screening and limited dilution subcloning. The monoclonal antibody was assayed by serological test and was confirmed by flow cytometry (FCM). 【Results】 From the cultural supernatant of D--peripheral blood transformed B lymphocytes, 3A6-C6, which agglutinated with R1R1(DCe/DCe)O-type RBCs but not with R2R2(DcE/DcE)O-type RBCs, was screened and preliminarily identified as anti-C. We established a hybridoma cell line secreting anti-C immunoglobulin M from B cells of D--individual successfully after hybridization with SHM-D33 myeloma cells. 【Conclusion】 The study had laid the groundwork for future research and development of human monoclonal antibodies against Rh antigens of RBC in future for diagnosis and screening purpose.

Objective To evaluate interventional techniques for the management of splanchnic artery areurysms. MethodsThere were 11 cases including splenic artery aneurysm in 5 cases, gastroduodenal in 5, and superior mesenteric artery (SMA) aneurysm in 1. Five patients presented with ruptured pseudoaneurysms and bleeding. Three patients presented with abdominal pain and epigastric pulsatile mass. Three patients were asymptomatic. Transcatheter endovascular coil embolization was used in 10 patients. Combination of the endovascular embolization and stent-graft placement in the SMA was performed in 1 patient with SMA areurysm. ResultsTechnical success was achieved in all cases with no significant complications. Hemostasis was achieved in all 5 patients. One patient was cured by combination of stenting and endovascular coil embolization. An average follow-up of 25.5 months in 11 cases revealed no complications. Doppler ultrasound scan showed complete thrombosis and occlusion of these aneurysms. ConclusionsTranscatheter endovascular interventional techniques are safe and effective methods for the treatment of splanchnic artery areurysms.