Objective: To analyze the frequency and investigate the causes of unexpected antibodies in D-negative blood donors. Methods: From January 2022 to December 2024, 3 768 D-negative blood donors sent to our laboratory were selected as research subjects. D-negative confirmation test and RhCE phenotype detection were applied by saline tube method and microcolumn gel indirect antiglobulin test (IAT), respectively. Antibody screening and identification were performed using the polybrene method and IAT column agglutination methods. Anti-D, anti-C and anti-G specificities were identified by a two-step adsorption-elution method, and the genotypes of D-negative samples were determined by RHD gene amplification, Sanger sequencing, and PacBio Single Molecule Real-Time (SMRT) sequencing. Results: Among D-negative donors, ccee and Ccee phenotypes accounted for the highest proportion, 55.68% (2 098/3 768) and 29.56% (1 114/3 768), respectively, while CcEE and CCEe phenotypes were the least, with one case detected in each, accounting for 0.03% (1/3 768). A total of 165 cases with D variant phenotype were detected, and the proportion of D variant was 4.38% (165/3 768) in the donors detected by D-negative confirmation test. Antibody screening positive blood donors were identified in 93 cases with a proportion of 2.47% (93/3 768). Antibody specificity was determined in 84 blood donors, and 9 samples showed no clear specificity. Anti-D was detected most frequently (n=68), in which 6 of them were detected having multiple antibodies, anti-D + anti-C (n=2), anti-D + anti-G(n=1), and anti-D + anti-E(n=3). The other antibodies detected were anti-E (n=1), anti-M(n=9), anti-P1 (n=3), anti-Le (n=1), and anti-HI(n=2). Fourteen cases were detected with anti-D in serological D-negative donors with C+ or E+ phenotype, in which three of them were DVI type 3 individuals and 11 cases were D negative individuals. Conclusion: The incidence of unexpected antibodies was higher in D-negative blood donors than in the total donors, with anti-D being the most common. The data provide insights for prevention and monitoring hemolytic disease of the fetus and newborn (HDFN) caused by anti-D. To ensure the safety of blood transfusion, routine unexpected antibody screening for RhD-negative blood donors is recommended to prevent the use of unexpected antibodies positive plasma in the clinic.

[Objective] To investigate the functional differences and potential effects of chromatin spatial structure in patients with normal karyotype and complex karyotype multiple myeloma. [Methods] High-throughput chromosome conformational capture (Hi-C) analysis was performed on plasma cells of 1 case with 1q21 complex karyotype and 1 case with normal karyotype multiple myeloma, and the differences in three-dimensional genome structure between the two patients were analyzed, and the transcriptome characteristics of plasma cells were combined to investigate the differential features through gene functional enrichment. [Results] A/B switch occurred in 36% of the chromatin compartments in two cases, and 1 041 genes in patient with complex karyotype had B/A switch. About 3 500 topological association domains (TADs) were identified in each sample, and there was no significant difference. The number of loops identified in complex karyotype sample was 1 069, which was 1/6 of the normal sample, and there were significant differences in the number of three different types of loops, which to some extent reflected the loss of genome stability. Transcriptome analysis showed significant differences in expression profiles between the two patients, and a total of 6 150 differentially expressed genes (3 303 up-regulated genes and 2 847 down-regulated genes) were identified. [Conclusion] Compared with patient with normal karyotype, patient with 1q21 complex karyotype multiple myeloma exhibit significant changes in the spatial structure of plasma cell chromatin at different levels, which leads to changes in gene expression and activation of pathways related to cancer progression.

Objective To compare the safety and efficacy of emergency balloon dilation angioplasty with emergency stent implantation in treating patients with acute anterior tandem occlusion caused by atherosclerosis at the starting segment of the internal carotid artery.Methods A total of 91 patients with stroke caused by acute anterior tandem occlusion,who were admitted to the First Affiliated Hospital of Hainan Medical College and the Third Affiliated Hospital of Guangzhou Medical University of China within 24 hours after disease onset to receive treatment from January 2018 to October 2022,were enrolled in this study.The patients were divided into balloon dilation angioplasty group(balloon dilation group,n=51)and stent implantation group(stenting group,n=40).The basic clinical data were compared between the two groups.The modified thrombolysis in cerebral infarction(mTICI)grade 2b-3 was defined as a good recanalization.The postoperative 90-day modified Rankin scale(mRS)score of 0-2 points was defined as a good clinical prognosis.Results The good recanalization rate and postoperative 90-day good clinical prognosis rate in the stenting group were 70％and 60％respectively,which were higher than 60％and 52％respectively in the balloon dilation group,and the differences between the two groups were not statistically significant(P=0.361 and P=0.391 respectively).The incidences of symptomatic intracranial hemorrhage(sICH),asymptomatic intracranial hemorrhage(aSICH),and mortality in the stenting group were 10％,32.5％,and 22.5％respectively,which in the balloon dilation group were 11.8％,41.2％,and 17.7％respectively,and the differences between the two groups were not statistically significant(P=1.000,P=0.396,and P=0.564 respectively).Conclusion For the treatment of patients with acute anterior tandem occlusion caused by atherosclerosis at the starting segment of the internal carotid artery,both emergency balloon dilation angioplasty or stent implantation are clinically safe and effective.(J Intervent Radiol,2024,33:593-598)

Objective:To serologically and genotypically analyze the pedigree of a case with a new allele c.175_176insGA of B el subtype and preliminarily explore the molecular mechanism of weak expression of glycosyltransferase B. Method:In the descriptive study,a 23-year-old male voluntary blood donor and his family members were selected for the study. The ABO and Le blood types of the proband and his family members was identified by the test tube method. The agglutination inhibition test was applied to detect the B and H antigens in saliva, and the Sanger sequencing and PacBio (Pacific Bioscience) third-generation haplotype sequencing were performed on the study subjects to identify genotypes. Finally, Expasy software were applied to amino acid translation of DNA sequences and prediction of protein length after gene alteration. ORF finder was applied to predict alternative start codons as well as open reading frames of mRNA, and protein expression mechanisms were analyzed.Results:The proband and her sister were B el subtype, her mother was AB el subtype, her father was normal O type, and all members of the family were Le(a+b+) phenotype. Sanger sequencing results showed that a new allele of c.175_176insGA was found in exon 4 of the proband, her mother, and her sister. Third-generation haplotype sequencing detected the haplotypes of the family members, which revealed that the proband was ABO*O.01.02/ABO*BEL.NEW (c.175_176insGA), the father was ABO*O.01.02/ABO*O.01.02, the mother was ABO*A1.02/ABO*BEL.NEW (c.175_176insGA), and the sister was ABO*O.01.02/ABO*BEL.NEW (c.175_176insGA). Analysis of the protein expression mechanism indicated that although the new allele of ABO*BEL.NEW was presumed to cause a frameshift mutation and result in a premature stop codon p.Asp59Glu*fs20 in exon 5, encoding an inactive glycosyltransferase, an alternative start codon could be utilized to initiate translation of B el subtype functional glycosyltransferase. Conclusion:Expression of the new allele of B el subtype is associated with the translation of B el subtype glycosyltransferase initiated by alternative start codons.

Presynaptic dopaminergic PET imaging is a useful method for the diagnosis of parkinsonism. Based on the expert consensus on operation and clinical application of dopamine transporter brain PET imaging technology published in 2020, this paper further recommends the relevant elements of result interpretation of presynaptic dopaminergic PET imaging.

Objective:To explore risk factors for clinical onset in children with uncontrolled self-limited epilepsy with centrotemporal spikes (SeLECTS) managed by 2 anti-seizure medications (ASMs).Methods:A total of 112 children with SeLECTS who were diagnosed at the Department of Pediatric Neurology of the Third Affiliated Hospital of Zhengzhou University from January 2018 to May 2021 were retrospectively reviewed.All of them were treated with conventional ASMs, and regularly followed up for 1-2 years.Types of therapeutic drugs, clinical seizure control status, presence of new seizure forms, electroencephalogram (EEG) were reviewed at follow-up visits.According to whether the seizures were controlled after the use of no more than 2 ASMs, patients were divided into poor response group (43 cases) and good response group (69 cases), and their clinical data and EEG characteristics were compared.Multivariate Logistic regression analysis was used to explore the risk factors for seizures that were uncontrolled by 2 ASMs. Results:There were significant differences in the age of onset ( χ2=8.919, P=0.003), seizure form ( χ2=4.218, P=0.040), seizure frequency ( Z=-7.664, P<0.001), EEG background slowing ( χ2=10.284, P=0.001), emergence of electrical status epilepticus during slow-wave sleep (ESES)( χ2=11.921, P=0.001), discharge generalization ( χ2=25.377, P<0.001), and presence of epileptic encephalopathy with spike-and-wave activation in sleep (EE-SWAS)( χ2=54.334, P<0.001) between groups.Multivariate Logistic regression analysis showed that seizure frequency ( P<0.001, OR=0.086, 95% CI: 0.022-0.329), discharge generalization ( P=0.006, OR=9.942, 95% CI: 1.918-51.527) and EEG background slowing ( P=0.041, OR=6.648, 95% CI: 1.077-41.038) were the 3 main risk factors associated with poor response to short-term medications of ASMs. Conclusions:Seizures are easily controlled in most SeLECTS patients medicated with ASMs with a favorable prognosis.Seizure frequency, discharge generalization and EEG background slowing are risk factors for the poor response to short-term pharmacotherapy in children with SeLECTS.

ObjectiveTo investigate the distribution of traditional Chinese medicine （TCM） syndrome types and influencing factors of protein-energy wasting （PEW） in chronic kidney disease （CKD） undergoing maintenance hemodialysis （MHD）. MethodsAccording to diagnostic criteria， 164 patients with MHD were divided into PEW group and non-PEW group. The clinical data of all patients were collected， including general information such as gender， age， height and weight， disease characteristics such as course， cormobidity， and haemodialysis duration， laboratory indicators such as blood routine， liver function， renal function， electrolyte， blood lipid， grip strength， and the four examinations. Logistic regression analysis was used to find the influencing factors of PEW by taking the clinical indicators with significant differences between the two groups （P＜0.05） as the independent variables， diagnosis of PEW as the dependent variable， and normal values as the reference. ResultsOut of 164 patients with MHD， there were 96 （58.5%） cases in PEW group and 68 cases （41.5%） in non-PEW group. Compared to the non-PEW group，PEW group had increased age， ratios of bedrest， deep vein preservation， edema， and low grip strength， percentages of comorbidities type 2 diabetes， cardiovascular and cerebrovascular diseases，infections and anemia， and levels of alanine aminotransferase and permine amin aminotransferase， as well as decreased body mass index， self-care ratio，internal arteriovenous fistula， red blood cell count， hemoglobin， serum total protein， serum albumin levels （P＜0.05）. The PEW group had significantly higher frequency of poor appetite and digestion， abdominal distension， fear of cold and preference of warmth， weak breathing and fatigue， poor appetite， oliguria， nausea and vomiting than non-PEW group （P＜0.05）. The incidence of both yin and yang deficiency syndrome and damp-turbidity syndrome were significantly higher in the PEW group than the non-PEW group， while that of liver-kidney yin deficiency syndrome and stirring of wind syndrome were lower （P＜0.05）. Logistic regression analysis showed that low BMI （＜22 kg/m²）， inability to take care of oneself， low grip strength，low serum albumin （＜38 g/L）， infection， older age， fear of cold and cold limbs，and poor appetite were the risk factors of PEW in patients undergoing MHD （P＜0.05）. ConclusionThe root syndrome of MHD-PEW patients is both yin and yang deficiency， concurrent with damp-turbidity syndrome. Low BMI， low serum albumin， infection and older age may be the influencing factors of PEW in patients undergoing MHD.

OBJECTIVE To establish the index system of quality evaluation standard for pharmacist training in cough and wheeze pharmaceutical care （CWPC） outpatient department， and to provide a basis for the selection of CWPC pharmacist training teachers and the improvement of training plans. METHODS Based on Kirkpatrick model， using the Delphi method， a total of 15 experts from 13 tertiary hospitals in 10 provinces in China were consulted to establish the standard index system of the quality evaluation for CWPC pharmacists training. Analytic hierarchy process （AHP） was adopted to determine the weights of each indicator and quantify the index system according to the weights of indicators at all levels. RESULTS The coefficient of expert authority was 0.810， the judgment coefficient was 0.727， and the familiarity coefficient was 0.893. The Kendall coordination coefficient of each index was 0.308-0.687. The P values of χ2 test were all less than 0.05， which indicated that the degree of coordination of the experts was high. After two rounds of correspondence， Kirkpatrick model-based index system of quality evaluation standard for CWPC pharmacist training was determined. The index system included 4 first-level indexes （participants’ reaction layer， learning gain layer， behavior improvement layer， training outcome layer）， 12 second-level indexes （such as training needs， teaching methods， theoretical knowledge， practical skills， job abilities， patient benefits， etc.） and 44 third-level indexes （such as clear training objectives， core system of CWPC， special device operation and evaluation， professional knowledge related to the treatment of cough and wheeze patients， promoting the construction of CWPC， improving patient compliance）. CONCLUSIONS The constructed index system of quality evaluation standard for CWPC pharmacist training has a certain level of authority and scientificity， and provides a scientific theoretical basis for quality evaluation of CWPC pharmacist training.

Objective:To analyze the clinical features of bronchiectasis, in order to improve the diagnosis, treatment, and prognosis.Methods:The clinical data of 78 children with bronchiectasis at Qingdao Women and Children′s Hospital of Qingdao University from January 2010 to January 2020 were analyzed, including age, regional distribution, etiology, clinical manifestations, lung function characteristics, results of lung high-resolution CT, results of fiberbronchoscopy examination, treatment, and prognosis.Results:Among 78 children with bronchiectasis, there were 35 males(44.9%)and 43 females(55.1%), with a median age of 8.7(6.1, 9.0)years.There were 51 children in rural areas(65.4%)and 27 children in urban areas(34.6%). The incidence of children in rural areas was higher than that in urban areas.The top three causes of bronchiectasis were 40 (51.3%)cases after infection, nine cases(11.5%)of primary immunodeficiency and seven cases(9.0%)of inhalation.The main clinical manifestations were wet cough, expectoration, wheezing and repeated respiratory infection.High-resolution CT of the lungs showed 36(46.2%)cases with diffuse bronchiectasis and 37 (47.4%) cases with localized bronchiectasis.Forty-nine(62.8%) cases had abnormal pulmonary ventilation function.Sixty-one(78.2%)cases were treated with fiberoptic bronchoscopy, of which 49(80.3%)cases had "fishbone" changes.After anti-infection, fiberbronchoscopy examination, expectorant and physical therapy, 67 cases were discharged and followed up for more than one year.Among them, 55 cases had symptoms improved, 12 patients had recurrent respiratory tract infection, and three patients died.Conclusion:The clinical manifestations of bronchiectasis in children have no obvious specificity.Early diagnosis, identification of the cause and comprehensive management are critical to prognosis of bronchiectasis.

Objective:To investigate the effects of excessive fluoride exposure on astrocytes and the expression of glial fibrillary acidic protein (GFAP), in vitro and in vivo. Methods:(1) In vivo experiment: 24 SPF SD rats, half male and half female, were randomly divided into control and fluoride exposed groups according to sex and body weight, 12 rats in each group. Rats were fed with < 1 mg/L and 50 mg/L sodium fluoride solution prepared by tap water for 6 months, respectively. The expression levels of GFAP protein in rat brain tissue were measured by immunofluorescence, immunohistochemistry and Western blotting. (2) In vitro experiment: adult (6-month-old) rat cortical astrocytes were extracted and cultured in primary culture (4 mmol/L sodium fluoride solution for 24 h), and the astrocytes were identified by immunofluorescence, and GFAP mRNA and protein expression levels were detected by real-time fluorescence quantitative PCR and Western blotting, respectively, and astrocytes apoptosis and calcium ion content were detected by flow cytometry. Results:(1) In vivo experiment: the results of immunofluorescence, immunohistochemistry and Western blotting showed that the GFAP protein expression level in brain tissue of rats exposed to fluoride was higher than that of control group (0.440 ± 0.200 vs 0.250 ± 0.120, t =-5.93, P = 0.027; 0.270 ± 0.020 vs 0.240 ± 0.050, t =-4.87, P = 0.040; 1.017 ± 0.001 vs 0.486 ± 0.006, t =-52.48, P = 0.001). (2) In vitro experiment: GFAP positive cells were identified as astrocytes by immunofluorescence; GFAP mRNA expression level was higher in fluoride exposed group than that of control group by real-time fluorescence quantitative PCR (2.780 ± 0.120 vs 0.134 ± 0.005, t =-37.84, P = 0.001). The Western blotting results showed that the GFAP protein expression level was higher in fluoride exposed group than that of control group (2.76 ± 0.10 vs 1.38 ± 0.05, t =-20.44, P = 0.002). Flow cytometry results showed that the apoptosis rate of astrocytes was higher in fluoride exposed group than that of control group (%: 55.0 ± 1.0 vs 3.5 ± 0.6, t =-10.28, P = 0.009) and the calcium ion content was lower than that of control group (%: 54 ± 9 vs 72 ± 13, t = 4.64, P = 0.043). Conclusion:Excessive fluoride exposure causes increased GFAP expression in astrocytes in vitro and in vivo, promotes apoptosis, and affects calcium signaling pathways.