Objective To explore the characteristics of military training injuries in high-altitude troops and determine the possible impact of daily diet on these training injuries in order to provide theoretical reference for scientific training and medical service support for high-altitude troops.Methods A cross-sectional scheme was adopted in this study.A self-designed Military Training Injury Questionnaire for Plateau Troops was used to survey the officers and soldiers from resident high-altitude troops in July 2024 for their training injuries,daily diet,and other situations.The obtained data were statistically analyzed.Results Among the 3 655 participants,the incidence of military training injuries was 17.87%.The subject with highest incidence was physical training(45.94%),the most common season was winter(31.39%).The most common sites of injury were waist(28.48%),knees(22.21%),and ankles(18.07%),and the most common types were sprains(28.48%),chronic fatigue injuries(18.38%)and strains(12.25%).The intake amounts of coarse grains and potatoes,bean products,aquatic products and nuts were relatively low in the daily diet of high-altitude troops.Multivariate logistic regression analysis found intake of fruit(OR=0.625,95%CI:0.508～0.768,P<0.001)and of nut(OR=0.759,95%CI:0.654～0.879,P<0.001)were correlated with the occurrence of training injury.Conclusion The occurrence pattern of military training injuries in high-altitude troops in this survey is basically consistent with that of previous reports,but the incidence rate is slightly decreased.Regular consumption of fruit and nut may be protective factors for the occurrence of training injuries.

Objective: To explore the role of the c.598G>A mutation of the ITGB3 gene in the occurrence of fetal and neonatal alloimmune thrombocytopenia (FNAIT) through its expression in vitro. Methods: The platelet antibodies in the sera of the affected neonate and her mother were detected using commercial enzyme-linked immunosorbent assay (ELISA), solid-phase agglutination, flow cytometry and the gold standard monoclonal antibody-specific immobilization of platelet antigens (MAIPA). The common human platelet antigen (HPA) genotypes of the neonate and her parents were obtained using the HPA-SSP method. The presence of mutations was analyzed by sequencing the exons of the ITGB3 and ITGA2B genes. The target gene of ITGB3 was obtained by PCR amplification using the existing human platelet cDNA. The wild-type ITGB3 eukaryotic expression vector was constructed by TA cloning technology. The 598G>A mutant ITGB3 eukaryotic expression vector was obtained by point mutation, and the plasmid DNA was co-transfected with that of ITGA2B (αⅡb) into HEK293 cells. The transfected cells stably expressing GP Ⅱb/Ⅲa were screened and obtained. The expression of GP Ⅱb/Ⅲa in 598G>A mutant transfected cells and the presence of antibodies against this mutation in the serum of mother were detected by flow cytometry and MAIPA. Results: Antibodies against HLA-class Ⅰ and GP Ⅱb/Ⅲa glycoproteins were detected in the serum of the neonate's mother, and subsequent HLA antibody-specific testing confirmed the presence of antibodies against HLA-B 57∶01 and A 02∶05. ITGB3 sequencing showed that the neonate and her father carried the c.598G>A point mutation, which results in the change of glutamate to lysine at position 200. Antibodies against GP Ⅱb/Ⅲa glycoproteins were not detected using constructed c.598G>A mutant transfected cells reacted with the maternal serum. Conclusion: The in vitro expression and analysis of the ITGB3 c.598G>A mutation did not support a role for this mutation in the pathogenesis of FNAIT. The establishment of this method facilitates the discovery of new platelet low-frequency antigens, and provides a theoretical foundation for the detection of antibodies against platelet antigens associated with patients with adverse pregnancy and childbirth histories.

Objective: To analyze CD36 gene by PacBio Sequel Ⅱ the third-generation sequencing technology (TGS), including non-coding sequence, and to investigate the molecular mechanism of CD36 deficiency. Methods: Flow cytometry was performed in the southern Chinese population to detect the CD36 phenotype. Among them, 15 cases of CD36 type I deficiency, 15 cases of CD36 type Ⅱ deficiency, and 10 positive samples were selected. The TGS of the CD36 gene was performed and statistical analysis was conducted. Results: 40 samples (including 15 cases of type I deficiency, 15 cases of type Ⅱ deficiency, and 10 positive samples) were subjected by TGS of CD36 full-length sequences (except part of intron1). A total of 180 polymorphic loci were identified. Among them, 13 kinds were in the coding region, the rest were in non-coding region, with most mutations located in regulatory regions such as the 5′-UTR and 3′-UTR. Conclusion: The high polymorphism of CD36 non-coding regions, particularly in regulatory sequences, provides mechanistic insights into type Ⅱ CD36 deficiency.

Objective:To evaluate the impacts of 2 reduction sequences on the surgical treatment of Schatzker type Ⅳ tibial plateau fractures with coronal plane subluxation.Methods:A retrospective analysis was conducted of the 53 patients who had been treated for Schatzker type Ⅳ tibial plateau fractures with coronal plane subluxation at Department of Orthopaedics, Fuzhou Second General Hospital from June 2020 to October 2023. The patients were divided into 2 groups according to the 2 reduction sequences employed during surgery. In the observation group of 25 patients [16 males and 9 females with an age of (47.2±10.2) years], the coronal plane subluxation was first reduced and temporarily fixed, followed by reduction of the medial plateau, and fixation of both the medial and lateral plateaus. In the control group of 28 patients [13 males and 15 females with an age of (50.6±15.4) years], in the traditional manner, the medial plateau was first reduced and temporarily fixed, followed by reduction of the coronal plane subluxation and finally by fixation of both plateaus. Outcomes compared between the 2 groups included surgical duration, intraoperative blood loss, and fracture healing time. Additionally, postoperative outcomes were assessed on the second day after surgery using such parameters as medial proximal tibial angle, lateral plateau widening, rate of coronal plane subluxation, and Rasmussen score. Knee joint function was evaluated at the final follow-up.Results:No significant differences were found between the 2 groups in baseline characteristics, ensuring comparability ( P>0.05). The mean follow-up duration for all patients was (12.6±4.2) months. There were no significant differences in surgical duration, intraoperative blood loss, fracture healing time or medial proximal tibial angle between the 2 groups ( P>0.05). The observation group demonstrated a significantly higher success rate by Rasmussen score (100.0% [25/25]) than the control group (78.6% [22/28]), significantly smaller postoperative lateral plateau widening [(3.70±2.42) mm] than the control group [(5.79±3.38) mm], a significant lower rate of coronal plane subluxation [16.0% (4/25)] than the control group [42.9% (12/28)], a significantly higher Hospital for Special Surgery (HSS) knee score [(83.8±5.4) points] than the control group [(80.0±6.5) points] (all P<0.05). Conclusion:Compared to the traditional reduction sequence, the modified reduction sequence prioritizing the reduction of coronal plane subluxation during surgery for Schatzker type Ⅳ tibial plateau fractures with coronal plane subluxation leads to superior short-term outcomes, effectively reduced widening of postoperative lateral tibial plateau and a decreased rate of coronal plane subluxation.

Objective: To explore the role of the c.598G>A mutation of the ITGB3 gene in the occurrence of fetal and neonatal alloimmune thrombocytopenia (FNAIT) through its expression in vitro. Methods: The platelet antibodies in the sera of the affected neonate and her mother were detected using commercial enzyme-linked immunosorbent assay (ELISA), solid-phase agglutination, flow cytometry and the gold standard monoclonal antibody-specific immobilization of platelet antigens (MAIPA). The common human platelet antigen (HPA) genotypes of the neonate and her parents were obtained using the HPA-SSP method. The presence of mutations was analyzed by sequencing the exons of the ITGB3 and ITGA2B genes. The target gene of ITGB3 was obtained by PCR amplification using the existing human platelet cDNA. The wild-type ITGB3 eukaryotic expression vector was constructed by TA cloning technology. The 598G>A mutant ITGB3 eukaryotic expression vector was obtained by point mutation, and the plasmid DNA was co-transfected with that of ITGA2B (αⅡb) into HEK293 cells. The transfected cells stably expressing GP Ⅱb/Ⅲa were screened and obtained. The expression of GP Ⅱb/Ⅲa in 598G>A mutant transfected cells and the presence of antibodies against this mutation in the serum of mother were detected by flow cytometry and MAIPA. Results: Antibodies against HLA-class Ⅰ and GP Ⅱb/Ⅲa glycoproteins were detected in the serum of the neonate's mother, and subsequent HLA antibody-specific testing confirmed the presence of antibodies against HLA-B 57∶01 and A 02∶05. ITGB3 sequencing showed that the neonate and her father carried the c.598G>A point mutation, which results in the change of glutamate to lysine at position 200. Antibodies against GP Ⅱb/Ⅲa glycoproteins were not detected using constructed c.598G>A mutant transfected cells reacted with the maternal serum. Conclusion: The in vitro expression and analysis of the ITGB3 c.598G>A mutation did not support a role for this mutation in the pathogenesis of FNAIT. The establishment of this method facilitates the discovery of new platelet low-frequency antigens, and provides a theoretical foundation for the detection of antibodies against platelet antigens associated with patients with adverse pregnancy and childbirth histories.

Objective:To evaluate the impacts of 2 reduction sequences on the surgical treatment of Schatzker type Ⅳ tibial plateau fractures with coronal plane subluxation.Methods:A retrospective analysis was conducted of the 53 patients who had been treated for Schatzker type Ⅳ tibial plateau fractures with coronal plane subluxation at Department of Orthopaedics, Fuzhou Second General Hospital from June 2020 to October 2023. The patients were divided into 2 groups according to the 2 reduction sequences employed during surgery. In the observation group of 25 patients [16 males and 9 females with an age of (47.2±10.2) years], the coronal plane subluxation was first reduced and temporarily fixed, followed by reduction of the medial plateau, and fixation of both the medial and lateral plateaus. In the control group of 28 patients [13 males and 15 females with an age of (50.6±15.4) years], in the traditional manner, the medial plateau was first reduced and temporarily fixed, followed by reduction of the coronal plane subluxation and finally by fixation of both plateaus. Outcomes compared between the 2 groups included surgical duration, intraoperative blood loss, and fracture healing time. Additionally, postoperative outcomes were assessed on the second day after surgery using such parameters as medial proximal tibial angle, lateral plateau widening, rate of coronal plane subluxation, and Rasmussen score. Knee joint function was evaluated at the final follow-up.Results:No significant differences were found between the 2 groups in baseline characteristics, ensuring comparability ( P>0.05). The mean follow-up duration for all patients was (12.6±4.2) months. There were no significant differences in surgical duration, intraoperative blood loss, fracture healing time or medial proximal tibial angle between the 2 groups ( P>0.05). The observation group demonstrated a significantly higher success rate by Rasmussen score (100.0% [25/25]) than the control group (78.6% [22/28]), significantly smaller postoperative lateral plateau widening [(3.70±2.42) mm] than the control group [(5.79±3.38) mm], a significant lower rate of coronal plane subluxation [16.0% (4/25)] than the control group [42.9% (12/28)], a significantly higher Hospital for Special Surgery (HSS) knee score [(83.8±5.4) points] than the control group [(80.0±6.5) points] (all P<0.05). Conclusion:Compared to the traditional reduction sequence, the modified reduction sequence prioritizing the reduction of coronal plane subluxation during surgery for Schatzker type Ⅳ tibial plateau fractures with coronal plane subluxation leads to superior short-term outcomes, effectively reduced widening of postoperative lateral tibial plateau and a decreased rate of coronal plane subluxation.

Objective To analyze the structure of CD36,search the possible B cell epitopes and prepare multi-antigen peptides(MAP)with B cell epitopes,so as to provide a preliminary experimental basis for the preparation of CD36 antibod-ies using MAP with B cell epitopes.Methods The potential B cell epitopes of CD36 were analyzed by bioinformatics meth-ods,including physical and chemical properties,secondary structure,potential phosphorylation and glycosylation sites.Eight-branch MAP with CD36 B cell epitopes were synthesized by FMOC using polylysine as the core matrix.The purity of MAPs was analyzed by reverse high-performance liquid chromatography chromatography(RP-HPLC),and the molecular weight of MAPs was determined by mass spectrometry.Results CD36 is a stable and hydrophilic alkaline protein,with multiple phosphorylation and glycosylation sites and strong antigenicity,and its secondary structure is mainly characterized by irregular curls.Four potential B cell epitopes were obtained and 4 MAPs containing potential B cell epitopes were pre-pared.RP-HPLC analysis showed that the purity of the MAPs were above 85%,and the molecular weight of 3 MAPs was consistent with the expected theoretical molecular weight.Conclusion CD36 on platelet has strong antigenicity.MAPs con-taining CD36 B cell epitopes can provide the experimental basis for the preparation and related research of CD36 antibodies.

Antibodies against human CD36 are responsible for several immune-mediated disorders. The detection of anti-CD36 antibodies using the standard monoclonal antibody (mAb) immobilization of platelet antigens (MAIPA) assay is hampered by a high frequency of false-negative results, most likely due to competitive inhibition of the mAb used as the capture antibody. We generated a panel of mouse mAbs against CD36 and seven hybridomas (GZ-3, GZ-13, GZ-70, GZ-143, GZ-413, GZ-507, and GZ-608), which were selected for MAIPA assays, as they reacted with mouse and human CD36. Fourteen anti-CD36 sera were assayed; all of which showed a positive reaction in a PakPlus (Immucor GTI Diagnostics, Inc., Waukesha, WI, USA) ELISA-based screening (optical density: 0.257–2.292). When the reference anti-CD36 mAb FA6-152 was used in the MAIPA assay, only 6/14 (42.9%) sera displayed a positive reaction. In contrast, anti-CD36 antibodies were detected in 13/14 (92.9%) sera when GZ-70 and GZ-608 mAbs were used. This significant improvement resulted in the identification of anti-CD36 antibodies by an antigen capture assay. Since patient’s platelets possibly carrying rare native antigens are used, this method will facilitate the identification of new platelet antibodies against CD36 that are involved in immune-mediated thrombocytopenia and other diseases, such as transfusion-related acute lung injury.

【Objective】 To search compatible and suitable platelets for platelet transfusion refractoriness (PTR) patient caused by compound antibodies against HLA and CD36. 【Methods】 ELISA method was used to detect the antibody against platelet antigens and the specificity of HLA-I antibody in PTR patients. The specificity of HLA-I antibody and corresponding epitopes of patients were analyzed using MATCH IT! and HLA Matchmaker software. The HLA genotype of both donor and patient was obtained by HLA-SSO method. Compatible or suitable donor platelets for PTR patients were searched through cross-reactive group (CREG) of HLA-I and HLA epitope-matched approach (Eplet). The matching degree was identified using monoclonal antibody-specific immobilization of platelet antigens (MAIPA) and the platelet suspension immunofluores-cence test (PIFT). Finally, the transfusion effect was evaluated according to the corrected count increment (CCI). 【Results】 Compound antibodies against both CD36 and HLA-I antigens were detected in two PTR patients, and their phenotype of CD36 was conformed to be type I deficient. Through LSA testing, high-frequency of HLA -I antibodies was found in these two patients, and the panel reactive antibody in patients 1 and 2 was 56% (54/96) and 53% (51/96), respectively. According to HLA-CREG and Eplet matching strategies, one donor of grade C-matching with patient 1 and one donor of grade D-matching with patient 2 were screened from the CD36 deficiency donor bank, respectively. And the selected donors avoided the antigen of HLA-I antibody epitope. These results of MAIPA and PIFT also confirmed that no immune response was detected between the patient and the donor. And a CCI of >4.5 within 24 hour of transfusion of compatible platelets was obtained in patient 2. 【Conclusion】 For PTR patients caused by HLA and CD36 compound antibodies, a combination strategy including serological cross-matching, HLA-CREG and Eplet approach should be used to select the CD36 deficient donor platelets which evaded the antigen corresponding to HLA-I antibodies and had the compatible HLA epitopes.

【Objective】 To find the HLA-matched platelets from platelet donor registry and track the transfusion effect for aplastic anemia patients in pregnancy with platelet transfusion refractory (PTR) caused by anti-HLA, so as to support the childbirth and follow-up treatment of the patients. 【Methods】 Antibodies to HPA and HLA were detected by ELISA kit and Luminex, respectively. DNA of the patient and 523 platelet donors from the donor registry were extracted for high-resolution HLA genotyping. The Risk Factors Evaluation Software of PTR was used to select the ABO-compatible and HLA-matched donors, without HLA Eplets specific to the patient. After MASPAT cross matching, the patient was transfused with 1 U of platelets, and the 24h post-transfusion effect was recorded. 【Results】 Only anti-HLAⅠantibody was found in the patient serum, and the specificity Eplet was 65QIA, including HLA-B^*27∶08, B^*27∶05, B^*07∶02, B^*55∶01, B^*67∶01 and B^*54∶01; anti-HLA Ⅱ antibody was negative. The HLA genotypes of both the patient and donor were HLA-A^*02∶07, A^*11∶01, B^*46∶01, B^*46∶01, C^*01∶02, 01∶03, DRB1^*04∶05, DRB1^*0901, DQB1^*03∶03 and DQB1^*04∶01. The results of MASPAT matching were negative. HLA-matched platelets transfusion provided a satisfactory posttransfusion platelet responses in patients(1 before vs 33 ×10⁹/L after). A baby boy was delivered by cesarean section 4 weeks later, and the same donor was recruited due to the mother′s low Plt and bleeding trend. The 24h posttransfusion Plt (×10⁹ / L) rose from 5 to 37 after the secondary transfusion of 1U platelet. The vital signs of the mother and her baby were normal during the two-day follow up. 【Conclusion】 The establishment of blood donor registry and screening of HLA-matched donors is an effective approach to treat PTR caused by HLA antibodies in pregnancy complicated with aplastic anemia.