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 explore the regulatory mechanisms underlying the weak expression of ABO blood group antigens due to variants in the non-coding regions of the ABO gene. Methods:From June 2014 to October 2023, a total of 29 samples from the Taiyuan Blood Center and local hospitals, which were serologically identified as having weak ABO antigen expression without detectable coding region mutations, were selected for this study. Full-length ABO gene sequencing was performed using third-generation long-read sequencing technology (Pacific Biosciences) to obtain complete haplotype sequences of the ABO gene. Variants in the non-coding regions were compared and identified to infer their regulatory effects on weak antigen expression. The procedures followed in this study were in accordance with the ethical standards of the World Medical Association′s Declaration of Helsinki (2013 revision). The Medical Ethics Committee of Taiyuan Blood Center has granted an exemption from ethical review. Results:18 bp deletions in the -35 to -18 region of the promoter were identified in 7 samples. Variants in intron 1 (+ 5.8 kb) were detected in 7 samples, including ABO* A (28+ 5792_5793delCT (1 case) and ABO* B (28+ 5793T>C) located in the GATA binding region; ABO* B (28+ 5808C>T) (1 case) in the E-box region; and ABO* B (28+ 5875C>T) (4 cases) in the RUNX1 binding region. Nucleotide variants at splice sites were detected in 2 samples, namely ABO* B (C.98+ 1G>A) and ABO* B (C.204-2A>C). Conclusion:Variants in the non-coding regulatory sequences of the ABO gene are a significant factor contributing to weak ABO antigen expression. In clinical ABO sequencing, it is essential to screen not only the conventional coding regions but also the flanking sequences, introns, and splice sites of the ABO gene to facilitate precise blood transfusion.

The statement"Blood is the material carrier of spirit and qi"indicates that the generation of blood relies on the coordinated governance of spirit and qi,while simultaneously serving as their material foundation.Based on the physiological connection among"blood-spirit-qi",this article explores the pathogenesis of premature ovarian insufficiency(POI)accompanied by anxiety and depression.It proposes that essence and blood deficiency,premature exhaustion of Tian Gui(reproductive essence),and failure of blood to nourish the spirit form a pathological chain:"blood deficiency-spirit disturbance-POI with anxiety and depression".Blood stasis obstructing the uterine chamber,disorder of the chong and ren meridians,and failure of blood to carry the spirit give rise to the transformation:"blood stasis-spirit depression-POI with anxiety and depression".Liver qi stagnation and disruption of qi and blood further exacerbate the imbalance between blood and spirit,aggravating the disease progression.Based on this pathological analysis,the general treatment principle of"regulating blood,harmonizing spirit,and rectifying qi"is established.For patients with blood deficiency syndrome,treatment should focus on tonifying essence and blood,replenishing reproductive essence,and nourishing the spirit,selecting Guanyuan(CV4),Zusanli(ST36),and Sanyinjiao(SP6)as main acupoints.For patients with blood stasis syndrome,treatment should aim to regulate the chong and ren meridians,promote blood circulation,and calm the spirit,selecting Qichong(ST30),Zhongji(CV3),and Xuehai(SP10)as main acupoints.Simultaneously,the method of regulating the liver should be applied throughout the entire treatment process to soothe the liver,regulate qi,relieve depression,and calm the spirit,selecting Baihui(GV20),Taichong(LR3),and Ganshu(BL18)as main acupoints.Appropriate acupuncture techniques and methods should be chosen according to the patient's constitution and condition,providing new therapeutic ideas and approaches for clinical practice.

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 explore the regulatory mechanisms underlying the weak expression of ABO blood group antigens due to variants in the non-coding regions of the ABO gene. METHODS: From June 2014 to October 2023, a total of 29 samples from the Taiyuan Blood Center and local hospitals, which were serologically identified as having weak ABO antigen expression without detectable coding region mutations, were selected for this study. Full-length ABO gene sequencing was performed using third-generation long-read sequencing technology (Pacific Biosciences) to obtain complete haplotype sequences of the ABO gene. Variants in the non-coding regions were compared and identified to infer their regulatory effects on weak antigen expression. The procedures followed in this study were in accordance with the ethical standards of the World Medical Association's Declaration of Helsinki (2013 revision). The Medical Ethics Committee of Taiyuan Blood Center has granted an exemption from ethical review. RESULTS: 18 bp deletions in the -35 to -18 region of the promoter were identified in 7 samples. Variants in intron 1 (+5.8 kb) were detected in 7 samples, including ABO*A (28+5792_5793delCT (1 case) and ABO*B (28+5793T>C) located in the GATA binding region; ABO*B (28+5808C>T) (1 case) in the E-box region; and ABO*B (28+5875C>T) (4 cases) in the RUNX1 binding region. Nucleotide variants at splice sites were detected in 2 samples, namely ABO*B (C.98+1G>A) and ABO*B (C.204-2A>C). CONCLUSION Variants in the non-coding regulatory sequences of the ABO gene are a significant factor contributing to weak ABO antigen expression. In clinical ABO sequencing, it is essential to screen not only the conventional coding regions but also the flanking sequences, introns, and splice sites of the ABO gene to facilitate precise blood transfusion.
ABO Blood-Group System/genetics* ; Humans ; Alleles ; Promoter Regions, Genetic ; Haplotypes ; Introns

Objective To construct mouse models of vaccinia virus infection using C57BL/6N and BALB/C mice infected with vaccinia virus VR-1354(WR(NIH TC-adapted)),and to compare the differences between the models in the two mouse strains.Methods Vaccinia virus VR-1354 was amplified,concentrated,and titrated,according to conventional method.The median lethal doses(LD50)of vaccinia virus V R-1354 for C57BL/6N and BALB/C mice were determined,respectively,and mice from the two strains were infected with the respective LD50.Lung tissues from control and experimental mice were removed for hematoxylin and eosin staining on days 3,7,and 14,and changes in the virus load in the lung tissues were measured at the same times.Results The titer of the amplified vaccinia virus VR-1354 was 3×108 plaque forming units(PFU)/mL.The LD50 values in C57BL/6N and BALB/C mice were 2.5×103and BALB/C 2.8×103PFU,respectively.After infecting mice from both strains with 2.5× 103 PFU of vaccinia virus VR 1354,lung tissue lesions worsened with increasing infection time,but the symptoms of the mice tended to recover after day 7.The virus loads in lung tissues began to increase on day 3 in both strains,peaked on day 7,and then gradually decreased.Conclusions C57BL/6N and BALB/C mouse models infected with vaccinia virus VR-1354 were established successfully.The virus can cause obvious lesions in mouse lung tissues,including inflammatory cell infiltration and alveolar cavity fluid exudation.The degree of disease onset and susceptibility differed between the two mouse strains,with C57BL/6N mice showing higher susceptibility to this virus than BALB/C mice.

Objective: To identify the phenotypes, antibodies and explore the molecular mechanisms of a patient who carries antibodies to RBC high-frequency antigens and his family members. Methods: The antibody identification test was performed for the proband by serological methods, and targeted NGS was subsequently used to detect mutations that occurred in blood group genes. Blood samples were collected from the proband and his family members. Sanger sequencing was used to verify the mutation of the XK gene. The expression of Kell blood group antigens was detected by serological methods and flow cytometry. K cells were used to detect the antibody specificity of the proband. The morphology of red blood cells was detected by the scanning electron microscopy. The serum creatine kinase levels of the proband and his family members were analyzed by colorimetric methods. Results: The results of the antibody identification test suggested that the proband might have antibodies to high-frequency antigens. NGS results suggested a homozygous mutation (c. 107G>A) in exon 1 of the XK gene in the proband, resulting in a truncated XK protein. The Sanger sequencing results of the proband were consistent with the NGS results, and the mutation was not found in other family members. The expression of Kell blood group antigens of the proband was not found by serological methods and flow cytometry. The results of the antibody specificity test showed that the proband had anti-Km antibodies. Spike-like changes were identified on red blood cells, and serum creatine kinase level was elevated in the proband. Conclusion: In this study, the McLeod phenotype caused by homozygous mutation (c. 107G>A) of XK gene was identified in Chinese individuals for the first time by the phenotype and molecular mechanism studies. The results of genotyping and phenotyping suggested that the McLeod phenotype caused by the mutation was compatible with the phenotypes of McLeod and K .

BACKGROUND:Hyperbaric oxygen,as one of the emerging means of fatigue elimination,has been increasingly valued and applied in the field of sports.However,there are fewer studies on the effect of hyperbaric oxygen intervention on fatigue elimination after high-intensity interval training shock microcycle.OBJECTIVE:To investigate the effect of hyperbaric oxygen intervention on the elimination of exercise-induced fatigue in the high-intensity interval training shock microcycle,and to study the corresponding mechanisms in terms of blood biochemical markers and metabolomics.METHODS:Twenty male college students were recruited from the Capital University of Physical Education and Sports,and randomly divided into a control group(n=10)and a hyperbaric oxygen group(n=10).Both groups underwent high-intensity interval training shock microcycle training for 2 weeks,a total of 12 sessions,with the following specific training program:warming up at 50%of the maximum heart rate for 10 minutes,and then pedaling at 90%-95%of the maximum heart rate for 4 minutes,repeating the program for 5 sessions,with a rest period of 2.5 minutes in between sessions,and finally pedaling at 50%of the maximum heart rate for 30 minutes.Subjects in the control groups recovered naturally after training,and those in the hyperbaric group recovered from training with hyperbaric oxygen,60 minutes each,at a pressure of 131.722 kPa.Blood biochemical markers and metabolomics data were analyzed and rating of perceived exertion was performed before,during and at 1 and 3 days after the experiment.Oxidative stress indicators and fatigue monitoring indicators were analyzed by Pearson correlation analysis.RESULTS AND CONCLUSION:(1)Regarding exercise-induced fatigue indicators,the control group showed varying degrees of increase in blood uric acid,creatine kinase,interleukin 6 and the rating of perceived exertion after training,while the hyperbaric oxygen group exhibited minimal changes in blood uric acid,creatine kinase,interleukin 6 and the rating of perceived exertion after training.Additionally,blood uric acid,creatine kinase,and interleukin 6 levels in the control group were significantly higher than those in the hyperbaric oxygen group at 1 day after training.(2)In the control group,superoxide dismutase levels decreased,while malondialdehyde levels increased after training.Conversely,in the hyperbaric oxygen group,superoxide dismutase levels increased,while malondialdehyde levels decreased after training.(3)Superoxide dismutase levels showed a negative correlation with blood uric acid,interleukin 6 and the rating of perceived exertion,while malondialdehyde levels exhibited a positive correlation with interleukin 6 and the rating of perceived exertion.(4)In the metabolomics analysis,significant changes were observed in the metabolic pathways of arachidonic acid metabolism and oxidative phosphorylation.Differential metabolites enriched in these pathways included arachidonic acid,prostaglandin D2,leukotriene D4,etc.To conclude,the high-intensity interval training shock microcycle induces oxidative stress,leading to exercise-induced fatigue in the body.Hyperbaric oxygen intervention can partially ameliorate oxidative stress levels and cause arachidonic acid metabolism and oxidative phosphorylation,thereby reducing oxidative damage,regulating inflammatory responses,promoting tissue repair,and alleviating exercise-induced fatigue.

Objective To construct mouse models of vaccinia virus infection using C57BL/6N and BALB/C mice infected with vaccinia virus VR-1354(WR(NIH TC-adapted)),and to compare the differences between the models in the two mouse strains.Methods Vaccinia virus VR-1354 was amplified,concentrated,and titrated,according to conventional method.The median lethal doses(LD50)of vaccinia virus V R-1354 for C57BL/6N and BALB/C mice were determined,respectively,and mice from the two strains were infected with the respective LD50.Lung tissues from control and experimental mice were removed for hematoxylin and eosin staining on days 3,7,and 14,and changes in the virus load in the lung tissues were measured at the same times.Results The titer of the amplified vaccinia virus VR-1354 was 3×108 plaque forming units(PFU)/mL.The LD50 values in C57BL/6N and BALB/C mice were 2.5×103and BALB/C 2.8×103PFU,respectively.After infecting mice from both strains with 2.5× 103 PFU of vaccinia virus VR 1354,lung tissue lesions worsened with increasing infection time,but the symptoms of the mice tended to recover after day 7.The virus loads in lung tissues began to increase on day 3 in both strains,peaked on day 7,and then gradually decreased.Conclusions C57BL/6N and BALB/C mouse models infected with vaccinia virus VR-1354 were established successfully.The virus can cause obvious lesions in mouse lung tissues,including inflammatory cell infiltration and alveolar cavity fluid exudation.The degree of disease onset and susceptibility differed between the two mouse strains,with C57BL/6N mice showing higher susceptibility to this virus than BALB/C mice.

BACKGROUND:Hyperbaric oxygen,as one of the emerging means of fatigue elimination,has been increasingly valued and applied in the field of sports.However,there are fewer studies on the effect of hyperbaric oxygen intervention on fatigue elimination after high-intensity interval training shock microcycle.OBJECTIVE:To investigate the effect of hyperbaric oxygen intervention on the elimination of exercise-induced fatigue in the high-intensity interval training shock microcycle,and to study the corresponding mechanisms in terms of blood biochemical markers and metabolomics.METHODS:Twenty male college students were recruited from the Capital University of Physical Education and Sports,and randomly divided into a control group(n=10)and a hyperbaric oxygen group(n=10).Both groups underwent high-intensity interval training shock microcycle training for 2 weeks,a total of 12 sessions,with the following specific training program:warming up at 50%of the maximum heart rate for 10 minutes,and then pedaling at 90%-95%of the maximum heart rate for 4 minutes,repeating the program for 5 sessions,with a rest period of 2.5 minutes in between sessions,and finally pedaling at 50%of the maximum heart rate for 30 minutes.Subjects in the control groups recovered naturally after training,and those in the hyperbaric group recovered from training with hyperbaric oxygen,60 minutes each,at a pressure of 131.722 kPa.Blood biochemical markers and metabolomics data were analyzed and rating of perceived exertion was performed before,during and at 1 and 3 days after the experiment.Oxidative stress indicators and fatigue monitoring indicators were analyzed by Pearson correlation analysis.RESULTS AND CONCLUSION:(1)Regarding exercise-induced fatigue indicators,the control group showed varying degrees of increase in blood uric acid,creatine kinase,interleukin 6 and the rating of perceived exertion after training,while the hyperbaric oxygen group exhibited minimal changes in blood uric acid,creatine kinase,interleukin 6 and the rating of perceived exertion after training.Additionally,blood uric acid,creatine kinase,and interleukin 6 levels in the control group were significantly higher than those in the hyperbaric oxygen group at 1 day after training.(2)In the control group,superoxide dismutase levels decreased,while malondialdehyde levels increased after training.Conversely,in the hyperbaric oxygen group,superoxide dismutase levels increased,while malondialdehyde levels decreased after training.(3)Superoxide dismutase levels showed a negative correlation with blood uric acid,interleukin 6 and the rating of perceived exertion,while malondialdehyde levels exhibited a positive correlation with interleukin 6 and the rating of perceived exertion.(4)In the metabolomics analysis,significant changes were observed in the metabolic pathways of arachidonic acid metabolism and oxidative phosphorylation.Differential metabolites enriched in these pathways included arachidonic acid,prostaglandin D2,leukotriene D4,etc.To conclude,the high-intensity interval training shock microcycle induces oxidative stress,leading to exercise-induced fatigue in the body.Hyperbaric oxygen intervention can partially ameliorate oxidative stress levels and cause arachidonic acid metabolism and oxidative phosphorylation,thereby reducing oxidative damage,regulating inflammatory responses,promoting tissue repair,and alleviating exercise-induced fatigue.