Objective To investigate the division, prol iferation and differentiation abil ities of nestin+/GFAP+cell after spinal cord injury and to identify whether it has the characteristic of neural stem cells (NSCs). Methods Twelvemale SD rats, aged 8 weeks and weighing 200-250 g, were randomized into 2 groups (n=6 per group): model group inwhich the spinal cord injury model was establ ished by aneurysm cl ip compression method, and control group in which no processing was conducted. At 5 days after model ing, T8 spinal cord segment of rats in each group were obtained and the gray and the white substance of spinal cord outside the ependymal region around central tube were isolated to prepare single cellsuspension. Serum-free NSCs culture medium was adopted to culture and serum NSCs culture medium was appl ied to induce differentiation. Immunohistochemistry detection and flow cytometry were appl ied to observe and analyze the type of cells and their capabil ity of division, prol iferation and differentiation. Results At 3-7 days after injury, the model group witnessed a plenty of nestin+/GFAP+ cells in the single cell suspension, while the control group witnessed few. Cell count of the model and the control group was 5.15 ± 0.71 and 1.12 ± 0.38, respectively, indicating there was a significant difference between two groups (P lt; 0.01). Concerning cell cycle, the proportion of S-phase cell and prol iferation index of the model group (15.49% ± 3.04%, 15.88% ± 2.56%) were obviously higher than those of the control group (5.84% ± 0.28%, 6.47% ± 0.61%), indicating there were significant differences between two groups (P lt; 0.01). In the model group, primary cells gradually formed threedimensional cell clone spheres, which were small in size, smooth in margin, protruding in center and positive for nestin immunofluorescence staining, and large amounts of cell clone spheres were harvested after multi ple passages. While in the control group, no obvious cell clone spheres was observed in the primary and passage culture of single cell suspension. At 5 days after induced differentiation of cloned spheres in the model group, immunofluorescence staining showed there were a number of galactocerebroside (GaLC) -nestin+ cells; at 5-7 days, there were abundance of β-tubul in III-nestin+ and GFAP-nestin+ cells; and at 5-14 days, GaLC+ ol igodendrocyte, β-tubul in II+ neuron and GalC+ cell body and protruding were observed. Conclusion Nestin+/GFAP+ cells obtained by isolating the gray and the white substance of spinal cord outside the ependymal region around central tube after compressive spinal cord injury in adult rat has the abil ity of self-renewal and the potential of multi-polarization and may be a renewable source of NSCs in the central nervous system.
Objective To summarize 10-year orthopedic experiences in the management of hemophilic pseudotumor and to discuss an individualized treatment strategy centered on surgery. Methods The clinical data of 8 patients with hemophilic pseudotumor between June 2015 and May 2025 were retrospectively analyzed. All patients were male, aged 15-37 years, with a mean age of 27.3 years. Seven patients had hemophilia type A and 1 had hemophilia type B; all had severe hemophilia and no detectable inhibitor. The disease duration ranged from 2 months to 12 years, with a mean of 2.5 years. There were 7 soft-tissue pseudotumors and 1 osseous pseudotumor. Lesions involved the lower leg in 3 cases, the thigh in 1, the hip in 1, both the hip and thigh in 2, and both the knee and thigh in 1. Pseudotumor size ranged from 5 cm×5 cm×4 cm to 32 cm×19 cm×15 cm. Treatment modalities included conservative treatment in 1 case, pseudotumor excision in 5 cases, and transfemoral amputation in 2 cases. Perioperative factor replacement was administered according to hemophilia type and factor activity, followed by wound care, anti-infective treatment when needed, drainage management, and rehabilitation. ResultsFor 7 patients undergoing surgery, the operation time was 60-120 minutes (mean, 88.6 minutes), and the intraoperative blood loss was 150-700 mL (mean, 342.9 mL). Six patients received coagulation factor Ⅷ at doses of 25 400-40 000 U (mean, 32 600 U); 1 patient received coagulation factor Ⅸ at a dose of 35 000 U; the length of hospital stay was 15-30 days (mean, 22.1 days). All patients were followed up 2-10 years (mean, 3.9 years.). One patient developed a superficial wound infection, which resolved after sensitive antibiotics and dressing changes. One patient with a lower-leg soft-tissue pseudotumor healed after conservative treatment and had no recurrence. Among the 5 patients treated with pseudotumor excision, 1 patient with an osseous pseudotumor achieved satisfactory wound and bone healing after pseudotumor resection, allogeneic bone grafting, and internal fixation; 3 patients with soft-tissue pseudotumors had no recurrence during follow-up; and 1 patient with a giant hip soft-tissue pseudotumor developed recurrence with ulceration and massive bleeding at 2 years after surgery and eventually died after unsuccessful rescue at another hospital. The 2 patients who underwent amputation achieved wound healing and returned to daily life after prosthetic fitting and rehabilitation. ConclusionSurgery remains the main treatment for hemophilic pseudotumors that fail conservative treatment or present with progressive enlargement, ulceration and infection, or compression of vital structures. Adequate perioperative factor coverage, precise preoperative imaging, complete excision whenever feasible, and meticulous elimination of dead space are essential to reduce complications and recurrence.