In maintaining normal function and activation processes, glycolysis, lipid metabolism, and amino acid metabolism play key roles in the energy demand of platelets. In the resting state, platelets primarily rely on glycolysis and aerobic oxidation to generate energy. Upon activation, platelets preferentially utilize glycolysis, as it can more rapidly provide the required ATP. In addition to glycolysis, platelets can also utilize glycogen and fatty acids as additional energy sources. The ATP provided by fatty acid oxidation is crucial for platelet activation. Additionally, during platelet storage, distinctive changes in energy metabolism occur. In the early stages of storage, platelets primarily rely on glycolysis and the pentose phosphate pathway (PPP) to generate energy. In the mid-storage phase, there is an increase in tricarboxylic acid cycle (TCA) metabolism. In the later stages of storage, cellular metabolism gradually declines. The regulation and flexibility of these metabolic pathways play a critical role in the survival and function of platelets in different states.

Objective: To accurately determine the ABO blood group of samples exhibiting forward/reverse grouping discrepancies by combining first-generation (Sanger) and third-generation (long-read) sequencing technologies. Methods: Five samples with ABO forward/reverse grouping discrepancies were selected. Serological testing was conducted using automated blood typing instruments and the tube method. Genotyping was conducted using both Sanger and long-read sequencing technologies. Results: Sanger sequencing identified specific genetic mutations in two samples, with genotypes of ABO BA. 04/ABO O.01.01 and ABO B3.05/ABO O.01.02. Further analysis with long-read sequencing revealed specific mutations in the +5.8kb region of intron 1 (c.28+5885C>T and c.28+5861T>G) in three samples where mutations were not detected by Sanger sequencing. These mutations affect the expression of the ABO antigens and are likely responsible for the ABO subgroup phenotypes. Conclusion: The integration of Sanger and long-read sequencing technologies effectively identifies genetic variations causing ABO subtypes, providing a scientific basis for enhancing clinical transfusion safety and ensuring accurate blood group determination.

Objective: To accurately determine the ABO blood group of samples exhibiting forward/reverse grouping discrepancies by combining first-generation (Sanger) and third-generation (long-read) sequencing technologies. Methods: Five samples with ABO forward/reverse grouping discrepancies were selected. Serological testing was conducted using automated blood typing instruments and the tube method. Genotyping was conducted using both Sanger and long-read sequencing technologies. Results: Sanger sequencing identified specific genetic mutations in two samples, with genotypes of ABO BA. 04/ABO O.01.01 and ABO B3.05/ABO O.01.02. Further analysis with long-read sequencing revealed specific mutations in the +5.8kb region of intron 1 (c.28+5885C>T and c.28+5861T>G) in three samples where mutations were not detected by Sanger sequencing. These mutations affect the expression of the ABO antigens and are likely responsible for the ABO subgroup phenotypes. Conclusion: The integration of Sanger and long-read sequencing technologies effectively identifies genetic variations causing ABO subtypes, providing a scientific basis for enhancing clinical transfusion safety and ensuring accurate blood group determination.

Objective To prepare mouse monoclonal antibodies against the ectodomain of E2 (E2ecto) glycoprotein of Western equine encephalitis virus (WEEV). Methods A prokaryotic expression plasmid pET-28a-WEEV E2ecto was constructed and transformed into BL21 (DE3) competent cells. E2ecto protein was expressed by IPTG induction and presented mainly as inclusion bodies. Then the purified E2ecto protein was prepared by denaturation, renaturation and ultrafiltration. BALB/c mice were immunized with the formulated E2ecto protein using QuickAntibody-Mouse5W as an adjuvant via intramuscular route, boosted once at an interval of 21 days. At 35 days post-immunization, mice with antibody titer above 1×10⁴ were inoculated with E2ecto intraperitoneally, and spleen cells were fused with SP2/0 cells three days later. Hybridoma cells secreting specific monoclonal antibodies were screened by the limited dilution method, and ascites were prepared after intraperitoneal inoculation of hybridoma cells. The subtypes and titers of the antibodies in ascites were assayed by ELISA. The biological activity of the mAb was identified by immunofluorescence assay(IFA) on BHK-21 cells which were transfected with eukaryotic expression plasmid pCAGGS-WEEV-CE3E2E1. The specificity of the antibodies were evaluated with E2ecto proteins from EEEV and VEEV. Results Purified WEEV E2ecto protein was successfully expressed and obtained. Four monoclonal antibodies, 3G6G10, 3D7G2, 3B9E8 and 3D5B7, were prepared, and their subtypes were IgG2c(κ), IgM(κ), IgM(κ) and IgG1(κ), respectively. The titers of ascites antibodies 3G6G10, 3B9E8 and 3D7G2 were 10⁵, and 3D5B7 reached 10⁷. None of the four antibody strains cross-reacted with other encephalitis alphavirus such as VEEV and EEEV. Conclusion Four strains of mouse mAb specifically binding WEEV E2ecto are successfully prepared.
Horses ; Animals ; Mice ; Encephalitis Virus, Western Equine ; Ascites ; Immunosuppressive Agents ; Antibodies, Monoclonal ; Immunoglobulin M

【Objective】 To study the relationship between ABO subtype, para-Bombay blood group and genotype, so as to explore the possible molecular mechanism of these two blood groups, and provide accurate genetic detection targets and theoretical basis for the accurate identification of ABO blood group. 【Methods】 First, the serology of 24 200 patients with blood type identification in the Ruijin Hospital from February to December in 2022 were analyzed, as well as 10 ambiguous ABO samples from other hospitals(3 were suspected ABO subtype and 7 were suspected para-Bombay blood group). Then ABO subtypes and para-Bombay blood groups were directly sequenced or post-clonal sequencing was performed to analyze ABO, FUT1 and FUT2 gene sequences. 【Results】 Among the 24 200 patients underwent blood type identification, 7 cases of ABO subtypes were detected. Among the 10 ambiguous samples sent by other hospitals, 2 of ABO subtypes, 1 of normal type A, and 7 of para-Bombay blood type were detected. In total, we identified blood types as follows: 1) 9 ABO subtypes: Ael(AEL.02/O.01.02), AelB(AEL.05/B.01), three of B3(2 of B3.03/O.01.01, 1 of B3.03/O.01.02), B(A)(BA.02/O.01.01), ABweak(A1.02/BW.07), Bweak(BW.31 /O.01.02), A2Bweak(A2.05 /BW.31); 2) 7 para-Bombay blood group: ABm^h (FUT1^*01N.13/FUT1^*01N.13), 4 of Am^h (3 of FUT1^*01N.06/FUT1^*01N.13, 1 of FUT1^*01N.13/FUT1^*01N.13); two of Bm^h (FUT1^*01N.06 /FUT1^*01N.06, FUT1^*01N.06/FUT1^*01N.13), all of FUT2 of the 7 cases were FUT2^*01/FUT2^*01. 【Conclusion】 Clinical ABO blood group variant samples need to be identified in combination with serological and molecular biology to improve the accuracy of identification, thus providing a reference for safe blood transfusion, organ transplantation, and the prediction and prevention of fetal-maternal immune hemolytic disease.

Objective To examines the correlation between lifestyle factors and semen quality among sperm donors at Anhui human sperm bank.Methods Demographic and lifestyle data were collected from 1,222 volunteers who donated sperm between January 2021 and December 2023,and their association with semen quality was analyzed.Results Univariate chi-square analysis revealed significant associations between several lifestyle factors and abnor-mal semen parameters(P＜0.05),including non-student status,frequent masturbation,short-term abstinence,low exercise frequency,frequent staying up late,smoking,drinking,and short sleep duration.Moreover,multiva-riate logistic regression analysis demonstrated that non-student status,longer abstinence time,and insufficient sleep were linked to abnormal semen volume.Additionally,abstinence time,exercise frequency,staying up late,smok-ing,and sleep duration were significantly correlated with abnormal semen concentration and sperm motility(P＜0.05).Conclusion Analysis reveals a close relationship between semen quality and volunteers'lifestyles,inclu-ding factors such as abstinence time,staying up late,sleep duration,smoking,drinking,and exercise frequency.

Objective To analyze the characteristics of males with autologous sperm preservation(ASP)in Anhui human sperm bank,and to explore the future direction of ASP in human sperm bank.Methods The basic infor-mation of males applied for ASP in Anhui human sperm bank from January 2019 to December 2023 was retrospec-tively analyzed.Results During this period,there were 424 males applied for ASP.93.40％(396/424)came from Anhui Province,of which 46.46％(197/424)came from Hefei.The age range of them was 15 to 59 years old.66.04％(280/424)had a college degree or above.23.11％(98/424)were employees of public institutions or enterprises.26.89％(114/424)were unmarried and 89.39％(379/424)were childless.67.45％(286/424)patients applied for ASP because of assisted reproductive technology treatment.15.33％(65/424)patients did it due to tumors,among which testicular cancer,lymphoma,leukemia and seminoma were the main reasons.A total of 1 163 semen samples were saved,and 53 males had used their sperm.Conclusion Only a few people applied for ASP,and the characteristics of males with ASP can be used to further strengthen publicity for key groups,espe-cially cancer patients,so as to benefit more people with autologous sperm preservation.

Objective To investigate the serological and molecular biology characteristics of ambiguous blood types a-mong voluntary blood donors in Putian,and to file records for relevant donors for future reference in case of their own trans-fusion needs or emergency donations to others.Methods A total of 68 593 blood samples from voluntary blood donors in Putian Central Blood Station between January 1,2019 and August 31,2023 were collected and tested for blood types using serological methods.ABO gene(including promoters,enhancers and seven exons as well as their flanking sequences and in-tron 6)and FUT1 gene testing were performed on ambiguous blood types.3D models were constructed using Chimira and Py-MOL software to predict the impact of gene mutations on enzyme structure.Results A total of 16 ABO subtypes were identi-fied by serological methods,with the highest detection rate as the para-Bombay phenotype(0.73 per 10 000),followed by the cisAB phenotype(0.44 per 10 000).Gene analysis revealed 12 cases with known mutations(4 cases of FUT1.01N.06/FUT1.01N.06,1 case of FUT1 01W.08/FUT1.01N.06,2 cases of A2.08/B.01,1 case of BA.02/O.01.01,1 case of A3.07/O.01.01,3 cases of cisAB.01/B.01),and key mutations were not found in 4 cases(2 cases of A1.02/B.01,2 cases of A1.02/O.01.02).3D molecular model analysis revealed that both A3.07 and FUT1.01W.08 allele can lead to a decrease in ac-tivity of the corresponding glycosyltransferases,resulting in the emergence of subtypes.Conclusion The most common phe-notype causing discrepancies in ABO blood type testing among voluntary blood donors from Putian is the para-Bombay phe-notype,with the most common allele being FUT1.01N.06.

Pulmonary sarcomatoid carcinoma(PSC)is a rare and highly malignant tumor,which includes the follow-ing five pathologic types:pleomorphic carcinoma,spindle cell carcinoma,giant cell carcinoma,carcinosarcoma and pulmonary blastoma.The onset of PSC is occult with non-specific clinical symptoms and signs.The clinical manifestations include irritat-ing cough,bloody sputum,dyspnea,chest pain and so on,which are closely related to the growth and invasion site of the tumor.PSC tends to metastasize early,so most patients are already in local advanced stage or advanced stage with a median survival of 9 months at the time of hospital visit.A patient with primary PSC which led to 90%stenosis in central airway was treated by com-bined method of vascular and tracheoscopic intervention in our respiratory center.This treatment prolonged the patient's survival time and got a satisfactory effect at 19-month follow-up after surgery.Herein we report the case for clinical reference.

Background and objective As a new technique developed in recent years,bronchoscopic interven-tion therapy has the advantages of minimal invasion,high safety and repeatability.The aim of this study is to investigate the clinical characteristics of bronchopleural fistula(BPF)induced by surgeries for lung malignancies or benign diseases and the effect of bronchoscopic intervention therapy for BPF,so as to provide support for prevention and treatment of BPF.Methods Data 64 patients with BPF who were treated by bronchoscopic intervention in Respiratory Disease Cen-ter of Dongzhimen Hospital,Beijing University of Chinese Medicine from June 2020 to September 2023 were collected.Patients with fistula diameter ≤5 mm were underwent submucous injection of macrogol,combined with blocking therapy with N-butyl cyanoacrylate,medical bioprotein glue or silicone prosthesis.Patients with fistula diameter＞5 mm were im-planted with different stents and cardiac occluders.Locations and characteristics of fistulas were summarized,meanwhile,data including Karnofsky performance status(KPS),shortbreath scale(SS),body temperature,pleural drainage volume and white blood cell count before and after operation were observed.Results For all 64 patients,96 anatomic lung resections in-cluding pneumonectomy,lobectomy and segmentectomy were executed and 74 fistulas occurred in 65 fistula locations.The proportion of fistula in the right lung(63.5％)was significantly higher than that in the left(36.5％).Besides,the right inferior lobar bronchial fistula was the most common(40.5％).After operation,KPS was significantly increased,while SS,body tem-perature,pleural drainage volume and white blood cell count were significantly decreased compared to the preoperative values(P＜0.05).By telephone follow-up or readmission during 1 month to 38 months after treament,median survival time was 21 months.33 patients(51.6％)showed complete response,7 patients(10.9％)showed complete clinical response,18 patients(28.1％)showed partial response,and 6 patients(9.4％)showed no response.As a whole,the total effective rate of broncho-scopic intervention for BPF was 90.6％.Conclusion BPF induced by pulmonary surgery can lead to severe symptoms and it is usually life-threating.Bronchoscopic intervention therapy is one of the fast and effective therapeutic methods for BPF.