Objective To investigate whether modification with IL-21 and CCL19 enhances killing and tumor-infiltrating efficiency of NKP30 CAR-T cells in lung cancer.Methods The modified IL-21-CCL19 NKP30 CAR-T cells expressing IL-21 and CCL19 fusion gene was constructed based on NKP30 CAR-T cells and stimulated with CD3CD28 antibodies and IL-2.The immunophenotype and migration of the cells in the presence of IL-21 were investigated using flow cytometry and migration experiments.Lactate dehydrogenase(LDH)release and sphere formation assays were used to assess the killing and infiltration capabilities of CAR-T cells,and the secretion levels of IFN-γ,IL-21 and CCL19 were determined with enzyme-linked immunospot assay(ELISPOT)and ELISA.A zebrafish model bearing HCG-27 cell xenograft was established by microinjection of the tumor cells into the yolk sac followed 24 h later by injection of the immune cells at the same site,and the fluorescence signals were captured using a fluorescent microscopy.Results The NKP30 ligand B7H6,which was almost undetectable in normal tissues and blood cells,was highly expressed(over 90%)in lung cancer cells.Compared with NKP30 CAR-T cells and conventional T cells,IL-21-CCL19 NKP30 CAR-T cells exhibited stronger proliferative and migration capabilities with the formation of central memory T cells.The reduced expressions of CTLA4 and PD1 in the constructed cells resulted in enhanced killing efficiency against lung cancer cells accompanied by significantly increased production of IFN-γ,IL-21 and CCL19.In the zebrafish models,CAR-T cells exhibited stronger cytotoxicity and proliferative abilities than typical T cells,but these differences were not statistically significant between the two CAR-T cells.Conclusion Modification of NKP30 CAR-T cells with IL-21 and CCL19 facilitates their access into solid tumors for more effective tumor cell killing while producing a large number of memory T cells.

Objective To investigate whether modification with IL-21 and CCL19 enhances killing and tumor-infiltrating efficiency of NKP30 CAR-T cells in lung cancer.Methods The modified IL-21-CCL19 NKP30 CAR-T cells expressing IL-21 and CCL19 fusion gene was constructed based on NKP30 CAR-T cells and stimulated with CD3CD28 antibodies and IL-2.The immunophenotype and migration of the cells in the presence of IL-21 were investigated using flow cytometry and migration experiments.Lactate dehydrogenase(LDH)release and sphere formation assays were used to assess the killing and infiltration capabilities of CAR-T cells,and the secretion levels of IFN-γ,IL-21 and CCL19 were determined with enzyme-linked immunospot assay(ELISPOT)and ELISA.A zebrafish model bearing HCG-27 cell xenograft was established by microinjection of the tumor cells into the yolk sac followed 24 h later by injection of the immune cells at the same site,and the fluorescence signals were captured using a fluorescent microscopy.Results The NKP30 ligand B7H6,which was almost undetectable in normal tissues and blood cells,was highly expressed(over 90%)in lung cancer cells.Compared with NKP30 CAR-T cells and conventional T cells,IL-21-CCL19 NKP30 CAR-T cells exhibited stronger proliferative and migration capabilities with the formation of central memory T cells.The reduced expressions of CTLA4 and PD1 in the constructed cells resulted in enhanced killing efficiency against lung cancer cells accompanied by significantly increased production of IFN-γ,IL-21 and CCL19.In the zebrafish models,CAR-T cells exhibited stronger cytotoxicity and proliferative abilities than typical T cells,but these differences were not statistically significant between the two CAR-T cells.Conclusion Modification of NKP30 CAR-T cells with IL-21 and CCL19 facilitates their access into solid tumors for more effective tumor cell killing while producing a large number of memory T cells.

Objective:To improve clinicians' understanding of Multisystem Inflammatory Syndrome in Children (MIS-C) and to reduce missed and misdiagnosis.Methods:The clinical data of a patient with Multisystem inflammatory syndrome in children admitted to Maternal and Child Health Hospital of Hubei Province, Tongji Medical College, Huazhong University of Science and Technology in January 2023 were analyzed retrospectively, together with literature review.Results:A 10-year-old male patient was admitted to hospital with a fever for 5 days. This patient had a history of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection 1 month prior to admission; after admission, he continued to have recurring high fevers, accompanied by abdominal pain and Kawasaki disease like manifestations, shock, multiple organ function impairment, and a significant increase in inflammatory indicators. Finally, the patient was diagnosed with MIS-C, treated with intravenous immunoglobulin, glucocorticoids and aspirin, and discharged with clinical improvement.Conclusion:MIS-C was a novel syndrome with Kawasaki-like manifestations.Currently, the overall prognosis for MIS-C is good, but its pathogenesis is unclear and the therapeutic consensus remains limited. More studies are needed to clarify the best diagnosis and treatment methods and true prognosis.

Objective:To explore the fixation with S 2 alar iliac screws (S 2AI) for unstable injury to the pelvic posterior ring. Methods:The clinical data of 18 patients were analyzed retrospectively who had been treated for unstable injury to the posterior pelvic ring by S 2AI screw fixation at Division of Orthopaedics and Traumatology, Department of Orthopaedics, Nanfang Hospital, Southern Medical University and at Department of Orthopaedics and Traumatology, The First Affiliated Hospital to University of South China from September 2017 to December 2020. They were 8 males and 10 females, with an average age of 40 years (from 20 to 64 years). According to Tile classification, there were 3 cases of type B2, 10 cases of type C1 and 5 cases of type C2. Sacral nerve injury was complicated in 3 patients (2 cases of Gibbons grade Ⅱ and one of Gibbons grade Ⅲ). Short-segment S 1-S 2AI fixation was used in 6 cases, contralateral S 1-S 2AI fixation in one case, lumbar-pelvic fixation in 5 cases, and sacroiliac triangle fixation in 6 cases. The accuracy of S 2AI placement was evaluated by CT after operation according to the Shillingford method; the reduction quality of pelvic fractures and complications were documented. Results:All the 18 patients were followed up for 19.8 months (from 12 to 36 months). Bony union was achieved in all fractures. A total of 21 S 2AI were implanted in the 18 patients without intraoperative neurovascular injury. Postoperative CT found penetration into the iliac cortex by S 2AI in 2 cases. According to the Matta criteria, the fracture reduction was excellent in 10 cases, good in 7 and fair in one. Of the 3 patients with preoperative sacral nerve injury, 2 with grade Ⅱ injury recovered to grade Ⅰ and one with grade Ⅲ injury recovered to grade Ⅱ after operation. Superficial infection occurred at the incision site in 2 patients after operation, and complications such as protrusion, rupture or loosening of implants were observed in none of the patients. Conclusion:S 2AI fixation can be flexibly applied to various types of posterior pelvic ring injury and can provide the pelvic ring and the lumbosacral junction with rigid fixation, leading to a low complication rate.

Background::The calcineurin inhibitor (CNI)-based immune maintenance regimen that is commonly used after renal transplantation has greatly improved early graft survival after transplantation; however, the long-term prognosis of grafts has not been significantly improved. The nephrotoxicity of CNI drugs is one of the main risk factors for the poor long-term prognosis of grafts. Sirolimus (SRL) has been employed as an immunosuppressant in clinical practice for over 20 years and has been found to have no nephrotoxic effects on grafts. Presently, the regimen and timing of SRL application after renal transplantation vary, and clinical data are scarce. Multicenter prospective randomized controlled studies are particularly rare. This study aims to investigate the effects of early conversion to a low-dose CNI combined with SRL on the long-term prognosis of renal transplantation.Methods::Patients who receive four weeks of a standard regimen with CNI + mycophenolic acid (MPA) + glucocorticoid after renal transplantation in multiple transplant centers across China will be included in this study. At week 5, after the operation, patients in the experimental group will receive an additional administration of SRL, a reduction in the CNI drug doses, withdrawal of MPA medication, and maintenance of glucocorticoids. In addition, patients in the control group will receive the maintained standard of care. The patients’ vital signs, routine blood tests, routine urine tests, blood biochemistry, serum creatinine, BK virus (BKV)/cytomegalovirus (CMV), and trough concentrations of CNI drugs and SRL at the baseline and weeks 12, 24, 36, 48, 72, and 104 after conversion will be recorded. Patient survival, graft survival, and estimated glomerular filtration rate will be calculated, and concomitant medications and adverse events will also be recorded.Conclusion::The study data will be utilized to evaluate the efficacy and safety of early conversion to low-dose CNIs combined with SRL in renal transplant patients.Trial registration::Chinese Clinical Trial Registry, ChiCTR1800017277.

Objective:To evaluate the effectiveness of integrated pediatric training course of extracorporeal membrane oxygenation(ECMO)based on simulation teaching.Methods:The instructors of ECMO team of Children′s Hospital of Fudan University developed the teaching curriculum.Team members as a unit were recruited to study simulation-based courses, whose theoretical knowledge, skills and teamwork were assessed, and feedback from the trainees were gathered.Results:Since May 2018, 22 teams of pediatric specialized hospitals or general hospitals have taken part in our ECMO simulation, with totally 132 participants, including 45 ICU doctors(34.1%), 60 ICU nurses(45.5%), 23 thoracic surgeons(17.4%) and four anesthesiologists(3.0%). Twelve of them(9.1%)had a little ECMO experience.After training, the trainees had a high evaluation on each part of the course, with average score more than four, of which the theoretical score was lowest.Their self-evaluation on the theory, skills, teamwork and confidence in the implementation of ECMO had been significantly improved.Most(80.3%)of the trainees were confident to carry out ECMO in their local hospitals.The trainees′ baseline score of theoretical knowledge was 55.2±7.6, and increased to 67.1±7.3 after training( P<0.001). The average teamwork score of the 22 teams was 70.2±8.2, and the qualified rate was up to 86.4%.The most prominent skill problems in operation assessment were no albumin and blood priming(90.9%), non-skill problems were extremely anxious during the overall assessment(84.8%)of the participants, poor team work(74.2%), poor leadership(68.2%). For the feedback of the whole course, 97.0% and 94.7% of the trainees thought that integrated training and simulation teaching were the highlights of the course.At present, 13(59.1%)units have successfully developed ECMO technology after the course, and the number of children treated has been up to 83, with a survival rate of discharge of 54.2%.However, it was found that the skill level of some participants decreased about 2 weeks after the course. Conclusion:As a new attempt of ECMO teaching, this curriculum has achieved certain efficiency in both teaching and clinical practice.At the same time, our study also plays a role in promoting the development of ECMO in pediatrics.

Objective:To meet the demand of medical system for talents, the training of medical students' competency has become a new direction of medical education. This study aims to evaluate the effectiveness of training quality in medical graduates through the competency scale.Methods:Taking "attitude", "skill" and "knowledge" as the evaluation dimensions, the competency development was divided into four levels of "state", "explain", "apply" and "transfer", and we proposed the competence concept of "A.S.K.-SEAT" and formulated an evaluation scale. Questionnaires and behavior event interviews (BEI) were conducted in medical graduates of Shantou University in 2018. Reliability and validity of the questionnaire were evaluated and current situation of different competency items were analyzed.Results:A total of 155 questionnaires were collected with good reliability and validity, and 15 graduates participated in BEI. A total of 21 A.S.K. competency items (including five basic competency items and two discriminating competency items) and SEAT textual descriptions were finally established.Conclusion:A.S.K.-SEAT scale can provide valid references for the competency evaluation of medical graduates.

Objective:To explore the clinical effect of the fractional carbon dioxide laser in the treatment of contracture scars.Methods:A retrospective before-after self-control study was conducted. From December 2016 to April 2021, 16 patients (7 males and 9 females, aged 3-49 years) with contracture scars causing impaired function of the adjacent joint were admitted to Shanghai Ninth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. Eighteen joint scars of 16 patients were treated with fractional carbon dioxide laser every 2-3 months until the joint retained its normal range of motion or the effect plateaued. The treatment times of each patient were recorded. Before the first treatment and 6 months after the last treatment, the ranges of motion of the affected joint were measured in each patient, and the difference was calculated, meanwhile, the Vancouver Scar Scale (VSS) was used to evaluate the scar of each patient. In the treatment of 1 joint scar in each of 6 patients (totally 6 times of treatments), the ranges of motion of the affected joint before the current treatment, immediately after the treatment, and at the first follow-up after the treatment were documented, and the differences between the ranges before and immediately after the treatment as well as between the ranges before and at the first follow-up after the treatment were calculated. Adverse effects after the treatment in the treatment area were documented. At the last follow-up, a self-made questionnaire was used to collect the implementation status of the physical therapy and other scar management modalities during the treatment interval and follow-up period. Data were statistically analyzed with Wilcoxon rank sum test.Results:Eighteen joint scars in 16 patients received 2 (1, 3) times of fractional carbon dioxide laser treatment. The range of motion of the affected joint of 16 patients 6 months after the last treatment was 56.5 (39.0, 128.8)°, notably higher than 38.4 (22.9, 116.3)° before the first treatment ( Z=-3.724, P<0.01), showing a remarkable improvement by 17.4 (8.0, 24.1)°. The vascular distribution, softness, and thickness scores, and total score in VSS scoring of scars of 16 patients 6 months after the last treatment were significantly lower than those before the first treatment ( Z=-2.989, -3.762, -2.814, -3.739, P<0.01), with the most obvious improvement in softness. In 6 times of treatments, the range of motion of the affected joint immediately after treatment and at the first follow-up of (2.5±0.6) months after treatment were 156.2 (148.0, 164.2)° and 160.2 (156.7, 166.4)°, both notably higher than 151.4 (145.7, 155.3)° before treatment ( Z=-2.201, -2.201, P<0.05), showing a remarkable improvement by 9.1 (4.4, 13.0)° and 13.1 (8.0, 15.7)°, respectively. No adverse effects such as blisters, infection, or hypertrophic scar formation were observed in the treatment area of 16 patients after treatment. Most patients adopted physical therapy, compression, silicone gel or sheets, and braces during the treatment interval and follow-up period. Conclusions:The fractional carbon dioxide laser can soften the scar and increase the range of motion of the affected joint, which is suitable for the clinical treatment of mild contracture scars.

Objective By establishing the intervention mode of 12-week gait retraining (GR) (with the specific aim of changing the habitual running gait), to determine the changes of vertical ground reaction force (vGRF), kinematics and dynamics of hip, knee, ankle joints, as well as stiffness of lower limbs in running before and after GR, and to explore the influence of running posture transformation on impact force and lower limb biomechanics. Methods Vicon motion capture system and Kistler 3D force measurement platform were used to collect the GRF and marker track of 30 runners (15 in experimental group and 15 in control group) before and after GR with the minimalist shoes at a speed of 12 km/h±5%. Results A total of 17 subjects (9 in experimental group and 8 in control group) completed the GR. After GR, the maximum loading rate of both groups decreased significantly, and the maximum loading rate of experimental group was lower than that of control group. The foot strike angle in experimental group decreased significantly after GR, and the plantarflexion angle and hip joint angular extension velocity increased in both groups. The force moment of ankle joint increased in experimental group, and the stiffness of lower limbs was significantly improved in both groups. Conclusions A 12-week GR exercise intervention model was successfully established, with 78% conversion rate (from rearfoot strike to forefoot strike). GR can effectively avoid the peak of impact force, reduce the maximum loading rate, increase the lower limb stiffness, and thus reduce or even avoid the risk of running injury caused by impact force and may provide a possibility for the improvement of running economy.

Animals ; Cdc20 Proteins/metabolism* ; Cell Line ; Embryo, Mammalian/embryology* ; Embryonic Development ; Female ; Infertility, Female/metabolism* ; Mice ; Mutation ; Oocytes/metabolism*