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 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.

Critically ill patients are at high risk for hospital acquired infections, which can significantly increase the mortality rate and treatment costs for these patients. Therefore, in the process of treating the primary disease, strict prevention and control of new hospital infections is an essential component of the treatment for critically ill patients. The treatment of critically ill patients involves multiple steps and requires a concerted effort from various aspects such as theory, management, education, standards, and supervision to achieve effective prevention and control of hospital infections. However, there is currently a lack of unified understanding and standards for hospital infection prevention and control. To address this, in March 2024, a group of experts in critical care medicine, infectious diseases, and hospital infection from China discussed the current situation and issues of hospital infection control in the intensive care unit together. Based on a review of the latest evidence-based medical evidence from both domestic and international sources, 2024 Expert Consensus on Hospital Acquired Infection Control Principles in the Department of Critical Care Medicine was formed, aiming to provide a basis for the development of hospital infection prevention and control strategies in the field of critical care medicine.

Objective:To analyze the incidence and clinical phenotype of the concomitant extragenital malformations in the patients with female reproductive tract anomalies.Methods:A retrospective study was conducted using clinical data of hospitalized patients diagnosed with uterine, cervical, or vaginal malformations from January 2003 to December 2022 in Peking Union Medical College Hospital. The malformations were classified according to American Society for Reproductive Medicine müllerian anomalies classification 2021, and in each type, the incidence and specific manifestations of concomitant extragnital malformations were analyzed.Results:A total of 444 patients were included. The overall incidence of concomitant extragenital malformations was 43.5% (193/444), including urinary system, skeletal system, and other system malformations. Renal malformations on the obstructed side were present in all patients with oblique vaginal septum syndrome (100.0%, 78/78). The total incidence of concomitant extragnital malformations was as high as 8/11 in uterus didelphys, 43.5% (10/23) in unicornuate uterus, 33.6% (79/235) in Mayer-Rokitansky-Küster-Hauser syndrome, 18.8% (6/32) in septate uterus and 18.5% (12/65) in cervical agenesis. Urinary system malformations (30.6%, 136/444) and skeletal system malformations (13.5%, 60/444) were the most common concomitant malformations in all types, in which, unilateral renal agenesis and scoliosis were the most common.Conclusions:Urinary and skeletal system malformations are important features of female reproductive tract anomalies. Urologic ultrasonography and spinal roentgenogram are recommended for all patients with female reproductive tract anomalies.

Primary prostate signet ring cell carcinoma (PPSRCC) is one of the extremely rare malignant tumors in the male urogenital system, and its diagnosis mainly relies on pathological and immunohistochemical examination. Compared with typical prostate cancer, PPSRCC is characterized by more aggressive with less treatment response and poor prognosis. Current researches on the pathogenesis, clinical manifestations, pathological characteristics, diagnosis, treatment and prognosis of PPSRCC were reviewed.

Objective:To investigate the optimal serum antibody titer in acute stage for the diagnosis of Mycoplasma pneumoniae (MP) infection by passive agglutination, and to evaluate the clinical diagnostic value of different antibody titers.Methods:A cross-sectional study.Eighty-eight pairs of clinical serum samples were collected from children with MP infection treated at the Department of Pediatrics in Shengjing Hospital of China Medical University from December 2016 to February 2017 and Children′s Hospital of Baotou in November 2019.The four-fold change of the double serum specific antibody titer was used as the gold standard, and the receiver operating characteristic (ROC) curve was plotted.When detecting the single serum in acute stage, different antibody titers were used as positive criteria to evaluate their clinical application value in the diagnosis of MP infection and find the most appropriate serum antibody titer as the diagnostic cut-off value.Results:(1)When the serum specific antibody titer ≥1∶40 was used as the positive criterion, the sensitivity was 72.9%, the area under the ROC curve was 0.817, and the specificity was 87.5%, which might cause overdiagnosis.When the serum specific antibody titer ≥1∶160 was used as the positive criterion, the specificity was 97.5%, the area under the ROC curve was 0.775, and the sensitivity was 52.1%, which might cause missed diagnosis.When the serum specific antibody titer ≥1∶80 was used as the positive criterion, the sensitivity was 60.4%, the specificity was 97.5%, and the area under the ROC curve was 0.823, overall performing better compared with the said two criteria.(2)After the disease lasted at least 5 days, blood samples were collected.About 72.5% of the children had antibodies, and 60.0% of the children had antibody titers ≥1∶80.Conclusions:(1)When the passive agglutination method is used to detect MP infection, antibody titer ≥1∶80 is recommended as the diagnostic standard.However, in clinical practice, the diagnosis of MP infection depends on clinical and other laboratory test results.(2) It is appropriate to collect blood samples on 5-7 days of illness.If MP infection is clinically suspected, and an antibody titer of 1∶40 is also suggestive, it can perform cooperative diagnosis based on molecular biology lab results or retest at a shorter interval.

Objective:To compare the clinical outcomes of arthroscopic external tension band fixation versus open reduction and internal fixation in the treatment of greater tubercle fracture of the humerus.Methods:A retrospective cohort study was conducted on 55 patients with greater tubercle fracture of the humerus admitted to Taizhou Hospital of Zhejiang Province from September 2019 to June 2022, including 24 males and 31 females, aged 26-80 years [(61.7±10.5)years]. Out of them, 35 patients treated with open reduction and internal fixation (open reduction group), and 20 patients were treated with external anchor tension band under arthroscopy (arthroscopy group). The operation time, and the Visual Analogue Scale (VAS) score, American Shoulder and Elbow Surgeons (ASES) score, Constant-Murley score and shoulder active range of motion (anterior flexion, abduction and posterior extension) before operation, at 1 month after operation and at the last follow-up were compared between the two groups. Bone healing was observed in both groups at the last follow-up. Postoperative complications were compared between the two groups.Results:All the patients were followed up for 12-29 months [(16.9±4.0)months]. There was no significant difference in operation time between the two groups ( P>0.05). There were no significant differences in the VAS score, ASES score, Constant-Murley score and shoulder active range of motion between the two groups before operation ( P>0.05). The VAS score of the arthroscopy group was 3(2, 3)points at 1 month after operation, which was significantly lower than that of the open reduction group [4(3, 4) points] ( P<0.01). No significant difference was found in the VAS score at the last follow-up between the two groups ( P>0.05).The ASES scores of the arthroscopy group were (70.6±4.2)points and (90.2±3.7)points at 1 month after operation and at the last follow-up respectively, which were significantly higher than those of the open reduction group [(64.7±6.4)points and (87.5±4.9)points respectively] ( P<0.05 or 0.01). There was no significant difference in the Constant-Murley score between the arthroscopy group [(71.8±4.3)points] and the open reduction group [(70.9±5.3)points] at 1 month after operation ( P>0.05), while the Constant-Murley score of the arthroscopy group was (94.1±3.1)points at the last follow-up, which was significantly higher than that of the open reduction group [(89.2±4.7)points] ( P<0.01). At 1 month after operation and at the last follow-up, ranges of motion of the anterior flexion, abduction and posterior extension were (52.7±12.3)° and (140.0±16.9)°, (57.4±8.6)° and (125.0±14.3)°, and 16(15, 19)° and 25(20, 30)° in the arthroscopy group respectively, which were significantly higher than those in the open reduction group [(42.2±5.2)° and (110.9±14.0)°, (52.8±6.0)° and (103.7±11.7)°, and 10(10, 20)° and 16(15, 25)° respectively] ( P<0.05 or 0.01). At the last follow-up, it was found that bony union was achieved in both groups. There were no obvious complications such as incision infection or joint stiffnessin both groups. In the open reduction group, 2 patients had internal fixation failure within 1-3 months after operation but was treated with revision operation; 6 patients developed shoulder stiffness at 3-6 months after operation but had outpatient rehabilitation. The incidence rate of postoperative complications in the arthroscopy group [0%(0/20)] was significantly lower than that in the open reduction group [23%(8/35)] ( P<0.05). Conclusion:Compared with open reduction and internal fixation with plates and screws, arthroscopic external anchor tension band fixation in the treatment of greater tuberosity fracture of the humerus has the advantages of earlier pain relief, better shoulder functional improvement, better recovery of shoulder mobility, and fewer complications.

Objective:To detect expression levels of complement regulatory proteins CD55 and CD59 in the peripheral blood of patients with Stevens-Johnson syndrome (SJS) /toxic epidermal necrolysis (TEN), and to preliminarily analyze their potential roles in the occurrence of SJS/TEN.Methods:Hospitalized patients with SJS/TEN (SJS/TEN group) were collected from the Department of Dermatology of the First Affiliated Hospital of Anhui Medical University and the Second Affiliated Hospital of Anhui Medical University from December 2017 to December 2022. Meanwhile, patients with maculopapular exanthema (MPE) and healthy physical examinees were also collected and served as the mild group and healthy control group, respectively. Flow cytometry was performed to determine the proportions of CD4 + T lymphocytes and CD8 + T lymphocytes in peripheral blood mononuclear cells (PBMCs). The expression levels of inflammatory cytokines tumor necrosis factor (TNF) -α, interleukin (IL) -6, IL-17, IL-10, interferon (IFN) -γ, IL-2, and IL-4 were detected using flow cytometric bead array technology. The mRNA expression levels of CD55 and CD59 in PBMCs were detected by real-time fluorescence-based quantitative PCR (qRT-PCR). Flow cytometry was also performed to determine the protein expression of CD55 and CD59 on the surface of CD8 + T lymphocytes. Statistical analyses were carried out using one-way analysis of variance and Tukey's test. Results:Totally, 13 patients with SJS/TEN, 27 patients with MPE, and 40 healthy controls were collected. Among the SJS/TEN patients, there were 8 males and 5 females, with their age being 18 to 84 (47.15 ± 19.99) years, and disease duration being 7.74 ± 2.63 days. No significant differences were observed in the gender distribution or age among the 3 groups (both P > 0.05). The proportions of CD4 + T lymphocytes did not differ among the 3 groups ( F = 3.84, P = 0.051). The proportions of CD8 + T lymphocytes in the peripheral blood were significantly higher in the SJS/TEN group (25.60% ± 4.57%) than in the healthy control group (16.20% ± 6.28%; q = 4.59, P = 0.018). The expression levels of inflammatory cytokines TNF-α, IL-6, IL-10, IL-17, and IFN-γ were significantly higher in the SJS/TEN group than in the healthy control group and mild group (all P < 0.001). In addition, the mRNA expression of CD55 ( F = 9.46, P < 0.001) and CD59 in PBMCs ( F = 15.14, P < 0.001) was significantly lower in the SJS/TEN group than in the mild group and healthy control group. The protein expression levels of CD55 ( F = 51.51, P < 0.001) and CD59 ( F = 31.59, P < 0.001) on the surface of CD8 + T lymphocytes were also significantly lower in the SJS/TEN group than in the other two groups and the healthy control group, respectively. Conclusion:Complement regulatory proteins CD55 and CD59 were downregulated in SJS/TEN patients, which may be associated with the activation of CD8 + T lymphocytes and excessive inflammatory responses.

Humans ; Critical Illness ; Multiple Organ Failure/etiology*

Objective:To investigate the safety and efficacy of a combined anterior and posterior approach in total hip arthroplasty (THA) for fused/ankylosed hip.Methods:37 patients who underwent THA for fused/ankylosed hip from January 2015 to December 2020 were retrospectively analyzed, including 28 males and 9 females, with an average age of 47.9±12.0 years (range, 26-72 years). Etiologies included 23 cases of ankylosing spondylitis, 9 cases of infectious arthritis of the hip in youth, and 5 cases of traumatic arthritis after acetabulum or femoral neck fracture. All patients underwent THA with combined anterior and posterior approach. These factors include operation time, blood loss, amount of transfused blood, blood transfusion rate, preoperative and postoperative Harris score, postoperative range of motion of the hip, and perioperative complications, etc. were evaluated. Postoperative radiography of the hip was performed to evaluate acetabular abduction angle, anterior inclination angle, the prosthesis fixation, osteolysis and heterotopic ossification around the hip.Results:A total of 37 patients were enrolled. The mean operative time was 147.6±16.8 min (range, 129-190 min); the mean estimated blood loss (EBL) was 850.0±10.8 ml (range, 600-1,200 ml); the blood transfusion rate was 59% (22/37), and the mean blood transfusion was 420±45.0 ml (range, 0-800 ml). All patients were followed up for 4.2±0.9 years (range, 1.2-7.2 years). The average abductor angle of the acetabular was 43.7°±5.4° (range, 31°-55°), and the average inclination angle was 20.9°±6.7° (range, 10°-35°); the preoperative Harris score was 47.1±9.9 (range, 40-55) and the mean Harris score at the last follow-up was 83.4±12.4 (range, 75-90). The preoperative range of motion of the hip in all directions was 0°. Postoperative hip range of motion was good, with a mean hip flexion of 95.5°±12.2° (range, 80°-110°), mean extension of 10.5°±3.4° (range, -10°-25°), and mean abduction of 38.0°±8.2° (range, 10°-50°). Postoperative complications were minor, including 2 case with poor wound healing, 2 cases with paresthesia or tingling sensation in the anterior or anterolateral thigh, which returned to normal within 3 months after surgery, and no deep infection or dislocation occurred. The acetabular cup was in poor position in 2 cases and the femoral stem was varus in 1 case, but the prosthesis was stable and no treatment was needed. All the acetabular cups and femur stems were confirmed with bone ingrowth on the last follow-up radiographs, and one patient had heterotopic ossification (Brooker grade 1). No osteolysis or wear of the acetabular liner was observed.Conclusion:Combined anterior and posterior approach (Gibson posterolateral approach + modified Hardinge approach) in THA for fusion/ankylosed hip can fully expose the operative field and sufficiently release the soft tissue, and the function of hip recovered well postopratively.