AIM:To explore the production and cytotoxicity of the reactive oxygen species(ROS)induced by diallyl trisulfide(DATS)in HL-60 cells.METHODS:HL-60 cells were either treated with various doses of DATS alone,or DATS combination with apocynin,a specific NADPH oxidase inhibitor,or with antioxidant N-acetyl-L-cysteine(NAC)for 0,1,3,6,12 and 24 h,respectively.The intracellular ROS level was measured by flow cytometry.The activity of NADPH oxidase was evaluated by NBT reduction experiment.The content of both malondialdehyde(MDA)and the protein carbonyl were analyzed by spectrophotometer.RESULTS:The results from flow cytometry indicated that DATS significantly increased the intracellular ROS level in HL-60 cells(P

Objective To study the effect of caveolin-1 gene expression on the proliferation of human gastric adenocarcinoma cells,and to explore the possibility for its future usage in gene therapy.Methods The full-length caveolin-1 gene was stably transfected into the MGC803 cell line by lipofectin.The Pcl neo vector was transfected at the same time as mock control.The expression of caveolin-1 was detected by Western blot in both the caveolin-1 gene transfected MGC803 cells and the controls.The cell cycle was analyzed by flow cytometry.Results After transfected with caveolin-1,MGC803 cells significantly up-regulated the expression of caveolin 1 and extended their doubling time.The cell proliferation was inhibited and the cell cycle was arrested in the G_0/G_1 phase.Conclusion Caveolin-1 can inhibit the proliferation of MGC803 cells and induce cell cycle arrest in G_0/G_1 phase.

OBJECTIVETo explore the expression, characteristics and roles of macrophage colony-stimulating factor receptor (M-CSF-R) in human leukemia cell lines.

METHODSPeripheral blood mononuclear cells (PBMCs) collected from 3 healthy persons, cord blood mononuclear cells (CBMCs) collected from 5 healthy persons and 4 human myelomonocytic leukemia cell lines including J6-1, J6-2, K562 and HL-60 were studied by using ABC immunoperoxidaes assay, indirect immunofluorescene staining, flow cytometry, and Western blot.

RESULTSM-CSF-R was noticed to be localized in the cytoplasm, nucleus and at the membrane in 4 human leukemia cell lines; expression of M-CSF-R was not detected in normal human PBMCs without PHA stimulation. Human PBMCs stimulated by PHA expressed a low level of M-CSF-R. Frequencies of membrane bound M-CSF-R (M-CSF-mR) expression in J6-1, J6-2, K562 and HL-60 were 78.9%, 72.6%, 54.9% and 58.0% respectively. Frequencies of cytoplasm and nucleus associated M-CSF-R (M-CSF-cnR) were 52.3%, 44.3%, 28.0% and 65.3% respectively. One form of M-CSF-R with a molecular weight of 120 000 was detected both in the cytoplasm and nucleus of HL-60 cells. The half-life of M-CSF-cnR in leukemia cells mentioned above was longer than that of corresponding M-CSF-R in stimulated CBMCs, and the half-life of M-CSF-mR in leukemia cells was extended except that of M-CSF-mR in K562 cells. Both anti-M-CSF-R monoclonal antibody and recombinant human M-CSF soluble receptor could cause the growth arrest of HL-60 cell in G(0)/G(1) phase, and could inhibit the formation of colony of HL-60 cell in soft agarose.

CONCLUSIONSExpression of M-CSF-R in leukemia cells is heterogeneous. The accumulation of cellular M-CSF-R results in the low degradation rate of cellular M-CSF-R in leukemia cells, which could be a potential mitotic signal. Signal mediated by M-CSF-R is important and necessary for the growth of HL-60 cell.

Cell Line ; HL-60 Cells ; Humans ; Leukemia ; Leukocytes, Mononuclear ; metabolism ; Macrophage Colony-Stimulating Factor ; metabolism ; Receptor, Macrophage Colony-Stimulating Factor ; Tumor Cells, Cultured

Objective To evaluate the medial submuscular plating of the femur after limb lengthening and correction with frame in patients with limb length discrepancy and/or angular deformity.Methods A retrospective study was conducted involving 12 patients who had been referred to Department of Orthopaedic Trauma,Beijing Jishuitan Hospital from December 2009 to November 2016 for limb length discrepancy and/or angular deformity.They were 3 males and 9 females,with an average age of 23 years (from 14 to 32 years).Altogether 4 left and 8 right sides were involved.Of them,11 got their length discrepancy and/or angular deformity secondary to injury to distal femoral epiphyseal plate caused by trauma in their childhood and one had length discrepancy and angular deformity following unsuccessful conservative treatment of superior condylar fracture of the femur.The femoral shortening ranged from 3 to 11 cm (average,6.7 cm),and the varus or valgus deformity of the knee ranged from 8° to 18° (average,12°).First,a monolateral fixator was installed on the lateral femur.The limb was lengthened from 7 to 14 days after diaphysis osteotomy.Femoral supracondylar osteotomy was conducted again in patients with knee angular deformity after the lengthening reached the expected length and the deformity was corrected with the aid of external fixator.After medial submuscular plating via the femoral lateral approach using minimally invasive techniques,the external frames were removed.The time for plate removal,limb lengthening and correction of the varus or valgus deformity were documented.Results The mean follow-up was 31 months (from 23 to 43 months) for the 9 patients who had their plates still in situ,and 6.5 months (from 2 to 14 months) for the 3 patients who had subsequently their plates removed.None had blood transfusion.All underwent distraction osteogenesis of the femur to their preoperatively expected length,ranging from 3 to 9 cm (mean,6.2 cm).Both varus and valgus deformities were corrected.None developed a deep infection.The range of motion of the knee joint was similar to that before operation in all but one patient who had a range of motion 40° less than the pre-operative one.The time for external fixation averaged 91 days (from 46 to 113 days),with an external index of 22 d/cm.All patients were satisfied with their outcomes.Conclusion Medial femoral submuscular plating after limb lengthening and angular deformity correction with the external fixator on the lateral side is a useful technique for patients with femoral limb length discrepancy and/or angular deformity,significantly shortening the time for external fixation.

Objective To introduce the technique we invented to treat hypertrophic and oligotrophic nonunion of femoral shaft fracture after dynamization of intramedullary nail (IMN) and to report its preliminary results.Methods The data of 2 cases with hypertrophic nonunions of femoral shaft fracture and 1 case with oligotrophic nonunion following IMN dynamization who had been treated by the technique from March 2006 to July 2017 in Beijing Jishuitan hospital were retrospectively studied.There were 2 females and 1 male,aged 50,66 and 24 years old.2 parallel half pins were inserted from anterior to posterior at the trochanteric and condylar zones of the femur for antegrade intramedullary nailing patients,and from lateral to medial sides for retrograde intramedullary nailing patient respectively,then the pins were connected with monolateral limb reconstruction fixator.The fracture nonunion site was compressed by the fixator up to 10-15 mm.The holes at the end of intramedullary nail where their screws were removed for dynamization were locked again with 2 locking screws,and then the frame was removed.The patients are allowed to start their rehabilitation and fully bear their body weight 1 day after the operation.Results The three patients were followed up for 25,22,and 7 months after the surgery,respectively.The X-ray films showed the fracture healed at 7 months in two case,and at 12 months in one.One patient got her nail removed 25 months after the operation,feeling good 37 months after the removal.Conclusion This technique is an option to manage the hypertrophic and oligotrophic fracture nonunion of the femur after dynamization of IMN.It is mini-invasive to the nonunion site,easy to practice and allows early rehabilitation.

Objective: To discuss the result of treating tibia deformity with fixator assisted nailing technique (FAN). Methods: A total of 5 patients with 7 limbs of tibial deformity-were treated with FAN technique. Etiology: 2 patients with bilateral tibial deformity suffered from Ricket's disease, 2 patients were malunion after tibial fracture, 1 patient was congenital pseudoarthrosis of tibia. This is a retrospective study. The unilateral external fixator was mounted on the medial side of tibia, and thenthe minimal invasive osteotomy was performed. After the deformity was corrected, the intramedullary nail was inserted to fix the tibia. None of the patients need bone autograft. The pre-operation and post-operation medial proximal tibia angle (MPTA), mechanical axis deviation (MAD) and range of motion (ROM) were measured and analyzed. Results: All the 5 patients were followed-up for 12-60 months (average 32 months). The osteotomy site united in 3-5 months (average 4.5 months) post-operatively. According to Paley’s imaging scores, 4 patients were excellent and 1 patient was good. According to Paley's functional result scores, 5 patients were excellent; according to Paley’s bone results evaluation, 4 patients were excellent and 1 patient was good. We achieved desired post-operative MAD (from medial 15 mm-lateral 10 mm) in 6 limbs. The MPTA in 7 limbs was corrected to normal (84°-90°). The ROM was not significantly changed before and after operation. The average ROM of knee before operation was 125°(120°-135°), and average postoperative ROM was 120°(115°-130°), No deep infection or neurovascular injury occurred. All patients were satisfied with the method and results of the operation. Conclusion FAN technique combines the advantage of external fixation and intramedullary nail, and it is a good method to treat tibial deformity, the patient should be carefully evaluated and selected for applying this technique.