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 compare the preparation efficiency of mouse pox and mouse hepatitis antibodies between two substrains of BALB/c and Big-BALB/c (B-BALB/c) mice, and to provide a theoretical basis and reference for the selection of laboratory animals in the preparation of monoclonal antibodies inducedin vivo through hybridoma.Methods Individuals weighing more than 5% of the weight of normal animals at 4 weeks of age (the criterion for late selection is more than 10%) were selected from a population of conventionally bred BALB/c mice and bred individually, and a subline of B-BALB/c mice was prepared after 10 generations of selection. A total of 40 BALB/c mice and 40 B-BALB/c mice aged 10 to 11 weeks, half male and half female, were selected and inoculated with the mousepox monoclonal antibody hybridoma cell line G23 or the murine hepatitis monoclonal antibody hybridoma cell line Y15 pre-treated with liquid paraffin, respectively. Mice ascites containing monoclonal antibodies were obtained by in vivo induction. The antibody titer was tested by indirect ELISA. The mice were grouped based on the sub-strains, gender and inoculation type of hybridoma to analyze the ascites production, antibody titer and antibody production, and to evaluate the antibody preparation efficiency of the two BALB/c mouse sub-strains.ResultsAfter 10 generations of breeding, the body weight of 10-week-old male and female B-BALB/c mice increased by 22.3% and 12.8%, respectively, compared with BALB/c mice of the same age. Compared with BALB/c mice, B-BALB/c mice had better tolerance and adaptation to secondary ascites collection. Compared with BALB/c mice, the ascites production and antibody titer during the preparation of antibodies in B-BALB/c mice were significantly increased, especially in the hybridoma cell line G23 vaccination group (both P＜0.000 1) . After inoculation with the hybridoma cell lines G23 or Y15, the average antibody production of B-BALB/c mice (14.99×10⁴ U and 33.22×10⁴ U) was higher than that of BALB/c mice (5.33×10⁴ U and 19.31×10⁴ U) (both P＜0.01). After inoculation with hybridoma cell line G23, the average antibody production per unit body weight of B-BALB/c mice (0.55×10⁴ U/g) was higher than that of BALB/c mice (0.23×10⁴ U/g) (P＜0.000 1). And the antibody production per unit body weight of female B-BALB/c or BALB/c mice was higher than that of male B-BALB/c or BALB/c mice (bothP＜0.01).Conclusion B-BALB/c mice can be used as an alternative to BALB/c mice in the in vivo induction of monoclonal antibody preparation, which can achieve the purpose of reducing the number of experimental animals used, lowering the labor cost, and improving the efficiency of antibody preparation.

Infectious bone defect is bone defect with infection or as a result of treatment of bone infection. It requires surgical intervention, and the treatment processes are complex and long, which include bone infection control,bone defect repair and even complex soft tissue reconstructions in some cases. Failure to achieve the goals in any step may lead to the failure of the overall treatment. Therefore, infectious bone defect has been a worldwide challenge in the field of orthopedics. Conventionally, sequestrectomy, bone grafting, bone transport, and systemic/local antibiotic treatment are standard therapies. Radical debridement remains one of the cornerstones for the management of bone infection. However, the scale of debridement and the timing and method of bone defect reconstruction remain controversial. With the clinical application of induced membrane technique, effective infection control and rapid bone reconstruction have been achieved in the management of infectious bone defect. The induced membrane technique has attracted more interests and attention, but the lack of understanding the basic principles of infection control and technical details may hamper the clinical outcomes of induced membrane technique and complications can possibly occur. Therefore, the Chinese Orthopedic Association organized domestic orthopedic experts to formulate An evidence-based clinical guideline for the treatment of infectious bone defect with induced membrane technique ( version 2023) according to the evidence-based method and put forward recommendations on infectious bone defect from the aspects of precise diagnosis, preoperative evaluation, operation procedure, postoperative management and rehabilitation, so as to provide useful references for the treatment of infectious bone defect with induced membrane technique.

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.

Objective:To explore the impact of a modified blood management strategy on blood loss and transfusion rates during outpatient total hip arthroplasty(THA).Methods:The retrospective research was performed in a total of 125 patients (125 hips) who underwent outpatient primary THA from January 2019 to December 2021 at a medical center. According to whether a modified blood management protocol was used or not, all patients were divided into two groups. Group A was used the original perioperative blood management strategies (1 g tranexamic acid, intravenously, 10 minutes before skin incision), and group B was used the modified perioperative blood management strategy (on the basis of the original protocol, 2 g tranexamic acid was sprayed locally in the joint cavity before the incision was closed, 1 g tranexamic acid was injected intravenously 3 hours after surgery, and 1 g tranexamic acid was injected intravenously again on the first day after surgery). There were 52 cases in group A, including 32 males and 20 females, aged 58.5±9.8 years (range, 39-69 years), 13 cases were developmental hip dysplasia (Crowe I°-II°), 24 cases were avascular necrosis of the femoral head, 10 cases were hip osteoarthritis, 3 cases were ankylosing spondylitis involving hip joint, and 2 cases were femoral neck fracture. Among the 73 patients in group B, there were 43 males and 30 females, aged 55.8±10.4 years (range, 42-67 years), including 17 cases of developmental hip dysplasia (Crowe I°-II°), 32 cases of avascular necrosis of the femoral head, 16 cases of hip osteoarthritis, 7 cases of ankylosing spondylitis involving hip joint, and 1 case of femoral neck fracture. Intraoperative blood loss, transfusion, deep vein thrombosis (DVT) events (vascular ultrasound, 2w Postop.), the hemoglobin (Hb) drop, the hematocrit (Hct) drop and other complications were recorded.Results:After using the modified strategy, the intraoperative blood loss was significantly reduced (305.6±38.6 ml vs. 416.2±88.3 ml, t=9.51, P<0.001), and the drop of hemoglobin was significantly decreased (18.1±4.0 g/L vs. 22.3±5.8 g/L, t=4.97, P<0.001). The drop of Hct also decreased significantly (7.3%±0.7% vs. 9.6%±1.3%, t=10.21, P<0.001), and total blood loss decreased significantly (720.6±57.4 ml vs. 919.6±86.3 ml, t=15.49, P<0.001). The hidden blood loss was also significantly lower than that in group A (414.9±71.1 ml vs. 503.5±96.4 ml, t=5.91, P<0.001). One patient (in group A) developed intra-articular hemorrhage 2 h after surgery and was transferred back to the inpatient ward for treatment after transfusion. Three patients (2.4%, 1 in group A and 2 in group B) developed symptomatic anemia and were discharged successfully after conservative treatment. Calf muscular venous thrombosis occurred in 3 patients (2.4%), but no symptomatic deep vein thrombosis occurred in all patients. Conclusion:According to this retrospective research, the use of modified blood management strategy during outpatient THA can further reduce intraoperative blood loss, hidden blood loss and postoperative hematocrit drop, and does not increase the occurrence of perioperative thrombosis-related complications.

Objective    To evaluate the clinical effects of segmentectomy versus lobectomy under single utility port video-assisted thoracic surgery on inflammatory factors and immune cells in peripheral blood of non-small cell lung cancer patients, and to analyze the effect of changes of postoperative inflammatory factors and immune cells on the prognosis of the patients. Methods    The clinical data of 256 patients who underwent segmentectomy or lobectomy under single utility port video-assisted thoracic surgery for non-small cell lung cancer in the First Affiliated Hospital of Hebei North University from January 2016 to October 2020 were retrospectively collected. According to the operation method, they were divided into a segmentectomy group (126 patients with 79 males and 47 females at an age of 63.4±6.2 years) and a lobectomy group (130 patients with 91 males and 39 females at an age of 62.9±5.6 years). The change of inflammatory factors (C reactive protein, interleukin-6, interleukin-8, tumor necrosis factor-α) and immune cells (CD4+T cells, CD8+T cells and natural killer cells) were recorded and analyzed before operation (T0) and 1 day (T1), 3 days (T2), 7 days (T3), 1 month (T4) after the operation between the two groups. According to postoperative recurrence situations, they were divided into a recurrence group and a non-recurrence group, multivariate logistic regression analysis was used to analyze the relationship between the change of postoperative inflammatory factors, immune cells, and the prognosis of patients with non-small cell lung cancer. Results    (1) There was no statistical difference in sex ratio, underlying diseases, body mass index, levels of preoperative inflammatory factors or immune cells between the two groups (all P>0.05). (2) The changes of postoperative inflammatory factors in the segmentectomy group were significantly less than those in the lobectomy group at T1-T3 (all P<0.05), and the changes of postoperative immune cells in the segmentectomy group were significantly less than those in the lobectomy group at T1-T4 (all P<0.05). (3) The changes of postoperative inflammatory factors and immune cells on postoperative day 3 in the recurrence group were significantly more than those in the non-recurrence group (all P<0.05). (4) Multivariate logistic regression analysis showed that the changes of postoperative inflammatory factors and immune cells on postoperative day 3 may be the risk factors for postoperative recurrence and metastasis in patients with non-small cell lung cancer (all P<0.05). Conclusion    Single utility port video-assisted thoracic surgery segmentectomy for the treatment of non-small cell lung cancer can reduce the inflammatory response and protect body's immune function, and the change of postoperative inflammatory factors and immune cells in postoperative day 3 may be the risk factors for postoperative recurrence and metastasis in patients with non-small cell lung cancer.

Objective:Comparing with the external fixator technique, investigate the clinical effect of fixator-assisted plating technique for treatment distal femoral valgus deformity in adolescent.Methods:A retrospective analysis of the relevant data of 22 patients (25 limbs) with acute correction of distal femoral valgus deformities from July 2015 to June 2019, according to the difference of the final fixation, they were divided two groups. The fixator-assisted plating group, including 8 patients (9 limbs), 5 boys and 3 girls, 5 left and 2 right, 1 bilateral, the physis were closed in 4 cases and opened in 4 cases, age 14.04±1.99 years (11.7-18 years). The external fixator group, including 14 patients (16 limbs), 6 boys and 8 girls, 8 left and 4 right, 2 bilateral, the physis were closed in 9 cases and opened in 5 cases, age 13.33±1.88 years (10.1-16.5 years). Measuring the mechanical axis deviation (MAD) and the mechanical lateral distal femur angle (mLDFA) in full length standing AP view X-ray of the lower limb pre and post operation. Recording the changed of limb length discrepancy (LLD) before and after surgery and the knee range of motion at 6 weeks, 3 months, 6 months and the last follow up post operation. The Kolcaba comfort status scale was used to evaluate the comfort of two groups at 2 weeks, 6 weeks, 3 months, 6 months and the last follow up. Meanwhile recorded the healing time of osteotomy sites and the complications.Results:The difference in general information between the two groups was not statistically significant. The fixator-assisted plating group followed up mean 26 months (range, 12-40 months), the healing time were 4.00±0.66 months, the mLDFA mean 73.33°±4.12° before surgery and 87.89°±1.69° after surgery and there was significant difference ( t=10.582, P<0.05). The external fixator group followed up mean 36 months (range, 22-42 months), the healing time were 4.00±0.66 months, the mLDFA mean 73.31°±3.95° before surgery and 87.31°±1.54° after surgery and there was significant difference ( t=14.118, P<0.05). The MAD were in the normal range in all patients after surgery, and there were no significant difference about healing time of the osteotomy sites and postoperative mLDFA between the two groups ( t=1.514, P=0.150; t=0.845, P=0.411). Comparing with the knee function, the fixator-assisted plating group was better at 6 weeks, 3 months, 6 months after surgery but there was no difference at the last follow up. Also, the fixator-assisted plating group felt more comfortable at 2 weeks, 6 weeks, 3 months, 6 months and there was no difference at the last follow up. All the patients with external fixator have mild pin sites infection and there were no obvious complications in patients with plate. Conclusion:The fixator-assisted plating technique can accurately correct the valgus deformity with satisfactory healing of the osteotomy in distal femoral compare with the external fixator technique, but the patients feel more comfortable and the knee joint function recovers faster and no risk of pin site infection, the clinical results was satisfactory with fewer complication.

Objective:To analyze the risk factors of avascular necrosis (AVN) after closed reduction and spcia casting in treating developmental dysplaisa of the hip (DDH).Methods:The patients with DDH who received closed reduction in our department from January 2016 to December 2017 were retrospectively reviewed. The inclusion criteria included aged at reduction ≥6 months, achieving successful reduction, having complete data with more than 2 years follow-up. A total of 48 cases with 54 hips were included in the present study. Among them, there were 2 males and 46 females, 41 unilateral hips and 13 bilateral hips. The mean age at closed reduction was 16.4±3.8 months (range 6-24 months). The mean follow-up duration was 2.9±1.8 years (range 2.3-4.1 years). Closed reduction was conducted under general anesthesia followed with a spcia cast immobilization. The abduction angle of the cast was recorded. The stability of reduction was evaluated by Ramsey safety zone. The maximum abduction and re-dislocation abduction were recorded. The quality of reduction was evaluated by the medial gap and femoral head coverage on intraoperative arthrography and post-reduction MRI. AVN was diagnosed according to Salter criteria. The risk factors of AVN were analyzed by univariate and binary logistic regression analysis.Results:AVN occurred in 12 hips (22.2%) of 54 hips. International Hip Dysplasia Institute (IHDI) grade and the difference between maximum abduction and cast abduction (Max-Cast abduction) were related to the occurrence of AVN in univariate analysis. The incidence of AVN in hips of IHDI grade 4 (42.9%, 9/21) was significantly higher than that in hips of IHDI grade 3 (9.7%, 3/31) (χ 2=6.007, P=0.018). However, the hips of IHDI grade 3 and 2 (0%, 0/2) presented a similar incidence of AVN (χ 2=0.000, P=1.000). The Max-Cast abduction was -0.7°±5.9° in the AVN group and 6.1°±7.6° in the AVN group ( t=2.125, P=0.038). Finally, IHDI grade ( OR=8.256, P=0.015) and Max-Cast abduction ( OR=0.832, P=0.047) were both independent factors of AVN in multivariate analysis. Conclusion:Most of the hips with AVN are IHDI grade 4 after closed reduction for DDH. The abduction angle in a spica cast could not be significantly related to the occurrence of AVN. However, the risk of AVN might be increased when the cast abduction is close to or beyond the maximum abduction. Safe abduction in the cast should be 5 to 10 degrees less than maximum abduction at least.

Objective:To explore the method and curative effect evaluation of repairing and rebuilding the long segmental bone defect after resection of long bone malignant tumors in adolescents.Methods:Reviewed our experience in 10 patients. Among them, 4 were male and 6 were female. The age was 10.2 years old (range, 6-16 years old). Including 7 cases of osteosarcoma, 3 cases of Ewing's sarcoma; 2 cases of humerus tumors, 7 cases of femoral tumors, and 1 case of tibial tumors-who presented between April 2015 and October 2019, in which the proportion of bone defect account for 71.4%-89.3% of the total length of diseased bone after tumor resection, with an average value of 80.9%. Considering the epiphyseal growth of young people, on the premise of ensuring enlarged tumor resection, we use traditional fibula autograft and prosthesis combined with steel plate, 3D printing design to retain the super-prosthesis of the epiphysis, and 3D printing bone trabecular was processed for the sake of making it grow successfully into the binding part of prosthesis and bone as we expect, simultaneously, the design of the short-force arm retaining the bilateral epiphysis can assit the steel plate and interlocking screws to enhance the stability.Results:The operation was completed successfully, and there were no complications such as nerve and blood vessel injury during the operation. After follow-up of 4-8 months, with an average follow-up of 22.9 months. One patient's review of X-ray film after 2 years of the operation showed that the transplanted fibula had a sign of bone resorption, and the internal fixation was in it's original position without obvious abnormalities. Two cases' auxiliary steel plates had fractured within one and a half to two years after surgery. With replacing the fractured steel plates, the internal fixation was good until now. According to the evaluation standards of the International Society of Limb Salvation: 2 cases of humerus osteosarcoma were respectively scored 23 points, 26 points, an average of 24.5 points; 4 cases of femoral osteosarcoma and 3 cases of femoral Ewing's sarcoma respectively scored 26 points and 25 points, 19 points, 30 points, 27 points, 24 points, 28 points (average, 25.6 points); 1 case of tibial osteosarcoma scored 26 points; all obtained relative satisfying treatment results.Conclusion:The combination of 3D printing and traditional machining techniques to produce an individualized long segmental prosthesis that preserves the epiphysis which applicated in the limb salvage treatment of bone malignant tumors can achieve excellent therapeutic effect.

Objective: To summarize the clinical characteristics, treatment response and prognostic factors of rhabdomyosarcoma (RMS) in children. Methods: The clinical characteristics such as age at diagnosis, primary tumor site, tumor size, pathological type, clinical stage, and risk grouping of 213 RMS patients (140 males and 73 females) treated in Hematology Oncology Center of Beijing Children′s Hospital, Capital Medical University, from May 2006 to June 2018 were analyzed retrospectively. The clinical characteristics, overall survival (OS), event free survival (EFS) and prognostic factors of children treated with the Beijing Children′s Hospital-Rhabdomyosarcoma (BCH-RMS) regimen were analyzed. Survival data were analyzed by Kaplan-Meier survival analysis, and single factor analysis was performed by Log-Rank test. Results: The diagnostic age of 213 cases was 48.0 months (ranged 3.0-187.5 months), of which 136 cases (63.8%) were younger than 10 years old. The head and neck region was the most common primary site of tumor (30%, 64 cases), followed by the genitourinary tract (26.8%, 57 cases). Among pathological subtypes, embryonal RMS accounted for 71.4% (152 cases), while alveolar RMS and anaplastic RMS accounted for only 26.8% (57 cases) and 1.9% (4 cases), respectively. According to the Intergroup Rhabdomyosarcoma Study Group (IRS), IRS-Ⅲ and Ⅳ accounted for 85.0% (181 cases) of all RMS patients. In all patients, 9.4% (20 cases) patients were divided in to low-risk group, 52.1% (111 cases) patients in to intermediate -risk group, 25.8% (55 cases) patients in to high-risk group, and 12.7% (27 cases) patients in to the central nervous system invasion group, respectively. All patients with RMS received chemotherapy. The cycles of chemotherapy were 13.5 (ranged 5.0-18.0) for patients without event occurrence, while 14.2 (ranged 3.0-30.0) for patients with event occurrence. Among the 213 patients, 200 patients had surgical operation, of whom 103 patients underwent surgery before chemotherapy and 97 patients at the end of chemotherapy, 21 patients had secondary surgical resection. Radiotherapy was performed in 114 patients. The follow-up time was 23.0 months (ranged 0.5-151.0 months) . There were 98 patients with relapsed or progressed disease and 67 patients with death. The median time to progression was 10 months, of which 67 (68.4%) relapse occurred within 1 year and no recurrence occurred after follow-up for more than 5 years. The 3-year EFS and 5-year EFS were (52±4) % and (48±4) %, while the 3-year OS and 5-year OS were (65±4) % and (64±4) % by survival analysis. The 5-year OS of the low-risk, intermediate-risk, the high-risk were 100%, (74±5) %, (48±8) %, and the 2-year OS of the central nervous system invasion group was (36±11) % (χ²=33.52, P<0.01). The 5-year EFS of the low-risk, intermediate-risk, the high-risk were (93±6) %, (51±5) %, (36±7) % and the 2-year EFS of the central nervous system invasion group was (31±10) % (χ²=24.73, P<0.01) . Survival factor analysis suggested that the OS of children was correlated with age(χ²=4.16, P=0.038), tumor TNM stage (χ²=22.02, P=0.001), IRS group (χ²=4.49, P<0.01) and the risk group (χ²=33.52, P<0.01). Conclusions This study showed that the median age of newly diagnosed RMS patients was 4 years. The head and neck and the genitourinary tract were the most common primary origin of RMS. The OS was low in single-center RMS children. The median time to recurrence was 10 months, and recurrence was rare 3 years later.