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Objective:To compare the genotypes and phenotypes between the monozygotic twins via whole genome sequencing to further clarify the autosomal dominant inherited neurofibromatosis type 1 (NF1) variants related to congenital pseudarthrosis (CP).Methods:According to the diagnostic criteria of congenital tibial pseudarthrosis and the clinical diagnostic criteria of NF1, two pairs of monozygotic twins with NF1 were included. Both were female and only one of each pair had congenital pseudarthrosis. The other did not have congenital pseudarthrosis. Whole genome sequencing was performed using the peripheral blood of the two pairs of monozygotic twins. Customized bioinformatics analysis was then performed to identify single nucleotide variants (SNVs), short insertion deletion variants (InDel), copy number variants (CNVs), and structural variants (SVs). Classified the variants according to the American College of Medical Genetics and Genomics (ACMG) and ClinGen criteria. The germline variants within the monozygotic twins were compared to identify the CP patients' unique variants. The shared pathogenic or likely pathogenic germline variants between the unique variants in the CP patients from the twins were also analyzed. Further, the identified disease-causing variants were validated by Sanger sequencing in the family of the twins and their parents. Finally, the genotypes and phenotypes regarding the pathogenic variants of the NF1 gene among the twins were characterized. Results:Both the two monozygotic twins were identified pathogenic variants in the NF1 gene. One with c.3047_3048del (p.Cys1016SerfsTer4), and the other with c.4267A>G (p.Lys1423Glu). By Sanger sequencing validation in family quads, the two CP patients and their siblings harbored de novo heterozygous variants of the NF1 gene. In addition to the NF1 gene, no other genes were identified pathogenic or likely pathogenic variants uniquely in the CP patients compared with their twin sisters, as well as SVs and CNVs. In addition, by analyzing the rare and damaging variants in the two CP patients from the two twins, they had no overlapping genes against the SNVs, InDels, SVs, or CNVs. Conclusion:Whole genome sequencing revealed that both the two monozygotic twins with NF1 were detected pathogenic variants of gene NF1. No other pathogenic variants specific to the CP patients among the twins were identified. The two CP patients shared no other common genes from the detected likely pathogenic variants.
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Objective:To identify and analyze different proteins expression in the periosteum of congenital pseudarthrosis of the tibia (CPT) using tandem mass tags (TMT) proteomics.Methods:The samples were divided into three groups, namely CPT with neurofibromatosis type 1 (NF1) group (NF1-CPT group), CPT without NF1 group (nonNF1-CPT group) and control group (patients with open tibial fracture). A fold change ≥1.5 or ≤0.66 and P-value <0.05 was regarded as the threshold to screen differentially expressed proteins (DEPs). Subsequently, bioinformatics resources such as online tools DAVID and STRING were used to conduct GO annotation, KEGG pathways enrichment and protein-protein interaction (PPI) network with DEPs. Results:A total of 347 proteins differentially expressed in NF1-CPT group, 212 of which were up-regulated and 135 down-regulated. We identified 467 DEPs in nonNF1-CPT group, including 281 up-regulated and 186 down-regulated. Among of them, NF1-CPT group and nonNF1-CPT group shared 231 DEPs, except for HLA-DRB1 which increased in NF1-CPT group but decreased in nonNF1-CPT group. The remaining 230 DEPs showed the same expression trend in the two positive groups, including 117 up-regulated and 113 down-regulated. In particular, a total of 116 proteins were altered only in NF1-CPT group, including 94 up-regulated and 22 down-regulated. However, there were 236 proteins altered only in nonNF1-CPT group, including 164 up-regulated and 72 down-regulated. The results indicated that the pathogenesis of NF1-CPT was similar as nonNF1-CPT largely with a few differences. Finally, compared with nonNF1-CPT, there were 47 proteins changed 1.5-fold and P-value <0.05 in NF1-CPT group. Conclusion:The proteins expression in the periosteum of CPT is different from that of normal tibia. The expression of periosteal protein is also different between NF1-CPT and nonNF1-CPT. The present study will deepen our understanding of the pathogenesis of CPT in the protein level.
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Objective:To identify the risk factors for premature proximal femur physeal closure (PPC) in children treated surgically for femoral neck fractures.Methods:Data of 106 children with an open triradiate cartilage who were treated surgically for a femoral neck fracture were retrospectively analyzed. Age, gender, laterality, mechanism of injury, the type of fracture, initial displacement, time to reduction, fixation method, whether the implant crossed the physeal plate, reduction method, reduction quality and development of femoral head avascular necrosis (AVN) were collected. PPC of the proximal femur was assessed through postoperative 6-12 months radiographs.Results:A total of 106 patients with an open triradiate cartilage were followed up, with an average duration of 20.4±13.3 months (range, 6-86 months). The overall rate of PPC following paediatric femoral neck fractures treated surgically was 36.8% (39/106). Among the 39 patients with PPC, 25 were males and 14 were females; the average age at the time of injury was 9.7±3.6 years (range, 3-15 years); 23 patients were involved in left hips and 16 were in right; the mechanisms of injury included motor vehicle accident in 5 fractures, falling injury in 21, sports-related injury in 12 and other causes in 1 fracture; two hips were Delbet type I, and 26 hips and 11 hips were Delbet type II and III, separately; type II and III of initial displacement were involved in 26 and 13 patients, separately; the mean duration from injury to surgery was 3.3±2.8 d (range, 1-14 d); 2 hips were treated with Kirschner wires, 35 hips with screw fixation, and the remaining 2 hips underwent screw and plate fixation; 2 hips had the hardware crossing the proximal femoral growth plate, and the remaining 37 hips didn’t; 15 hips were treated by closed reduction and internal fixation, and the other 24 hips underwent open reduction and internal fixation; anatomical reduction was achieved in 14 patients, acceptable reduction in 24 hips and unacceptable reduction in 1 hip; 24 hips developed AVN at the latest follow-up and the remaining 15 hips didn’t. Statistical analysis indicated that age ( t=3.875, P< 0.001), the severity of initial displacement ( Z=-2.118, P=0.034) and the rate of AVN ( χ2=42.280, P< 0.001) in patients with PPC were significantly higher than those in patients without; Logistic regression analysis confirmed age ( OR=1.288, P=0.011) and AVN ( OR=40.336, P< 0.001) as risk factors for PPC. ROC curve analysis indicated 10 years was the cut off age to significantly increase the rate of PPC. The rate of PPC in patients aged over 10 years (63.6%, 21/33) was significantly higher than that (24.7%, 18/73) in those aged less than 10 years ( χ2=14.848, P< 0.001). Conclusion:Age over 10 years and AVN are risk factors for PPC in children with femoral neck fractures treated surgically.
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Objective To detect the expression of miR-373 in osteosarcoma cells and explore its effects on cell proliferation,invasion,and migration.Methods Human osteosarcoma cell line SJSA-1 and human osteoblast hFOB 1.19 cultured in vitro.Human osteosarcoma cell line SJSA-1 were randomly divided into four groups:blank control group (CK group),negative control group (NC group),miR-373 over-expressed plasmid group (miR-373 mimic group) and miR-373 inhibited plasmid group (miR-373 inhibitor group).Cells were transfected with FuGENE(R) HD transfection reagent.After 48 hours of transfection,the relative expression of miR-373 was detected by quantitative real-time polymerase chain reaction (qRT-PCR),cell proliferation by cell counting kit-8 (CCK-8) and cell invasion and migration by Transwell.Results The relative expression of miR-373 in SJSA-1 cells was significantly higher than that in hFOB-1.19 cells (P ＜ 0.05);compared with CK group and NC group,the relative expression of miR-373 in the miR-373 mimic group increased significantly (P ＜ 0.05),and the relative expression of miR-373 in the miR-373 inhibitor group was significantly lower than that in the control group (P ＜ 0.05);optical density (OD) value of miR-373 mimic group at 24 h,36 h,and 48 h were significantly higher than that of CK group and NC group (P ＜ 0.05),and OD value of miR-373 inhibitor group at 24 h,36 h,and 48 h were significantly lower than that of CK group and NC group (P ＜ 0.05);the number of cell migration in the miR-373 mimic group was significantly higher than that in the CK group and NC group (P ＜ 0.05),and the number of cell migration in the miR-373 inhibitor group was significantly lower than that in CK group and NC group (P ＜ 0.05);the number of cell migration in the miR-373 mimic group was significantly higher than that in the CK group and NC group (P ＜ 0.05),and the number of cell migration in the miR-373 inhibitor group was significantly lower than that in CK group and NC group (P ＜ 0.05).Conclusions The expression of miR-373 is up-regulated in osteosarcoma.Overexpression of miR-373 can promote the proliferation,invasion,and migration of osteosarcoma cells,while the low expression of miR-373 can inhibit the proliferation,invasion,and migration of osteosarcoma cells,which may be an important target for the diagnosis and treatment of osteosarcoma.
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Objective@#To detect the expression of miR-373 in osteosarcoma cells and explore its effects on cell proliferation, invasion, and migration.@*Methods@#Human osteosarcoma cell line SJSA-1 and human osteoblast hFOB 1.19 cultured in vitro. Human osteosarcoma cell line SJSA-1 were randomly divided into four groups: blank control group (CK group), negative control group (NC group), miR-373 over-expressed plasmid group (miR-373 mimic group) and miR-373 inhibited plasmid group (miR-373 inhibitor group). Cells were transfected with FuGENE® HD transfection reagent. After 48 hours of transfection, the relative expression of miR-373 was detected by quantitative real-time polymerase chain reaction (qRT-PCR), cell proliferation by cell counting kit-8 (CCK-8) and cell invasion and migration by Transwell.@*Results@#The relative expression of miR-373 in SJSA-1 cells was significantly higher than that in hFOB-1.19 cells (P<0.05); compared with CK group and NC group, the relative expression of miR-373 in the miR-373 mimic group increased significantly (P<0.05), and the relative expression of miR-373 in the miR-373 inhibitor group was significantly lower than that in the control group (P<0.05); optical density (OD) value of miR-373 mimic group at 24 h, 36 h, and 48 h were significantly higher than that of CK group and NC group (P<0.05), and OD value of miR-373 inhibitor group at 24 h, 36 h, and 48 h were significantly lower than that of CK group and NC group (P<0.05); the number of cell migration in the miR-373 mimic group was significantly higher than that in the CK group and NC group (P<0.05), and the number of cell migration in the miR-373 inhibitor group was significantly lower than that in CK group and NC group (P<0.05); the number of cell migration in the miR-373 mimic group was significantly higher than that in the CK group and NC group (P<0.05), and the number of cell migration in the miR-373 inhibitor group was significantly lower than that in CK group and NC group (P<0.05).@*Conclusions@#The expression of miR-373 is up-regulated in osteosarcoma. Overexpression of miR-373 can promote the proliferation, invasion, and migration of osteosarcoma cells, while the low expression of miR-373 can inhibit the proliferation, invasion, and migration of osteosarcoma cells, which may be an important target for the diagnosis and treatment of osteosarcoma.
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Objective To explore the risk factors related to refracture after union of congenital pseudarthrosis of the tibia with a combined surgical,and to provide an important reference for clinical treatment.Methods From April 2007 to August 2011,40 cases of congenital pseudarthrosis of the tibia (CPT) confirmed by the use of intramedullary nail combined Ilizarov external fixator treatment of bony union were obtained for retrospective analysis.There were 27 males and 13 females,and the average age was 3.6±2.7 years (ranged from 0.8 to 11.3 years),and the follow-up time are all over 36 months.Survival analysis were applied respectively on gender,whether the merger of neurofibromatosis type 1 (NF-1) and ankle joint,operative age,fibular status and the cross-sectional area ratio of healing site.Multifactorial Cox proportional hazards regression analysis was performed on the single factor with statistical significance.Results No significant inter-group differences existed in gender or whether combined NF-1.In operative age ≥3 years group whose refracture-free survival was significantly better than those ＜ 3 years.Intact fibula or stabilised by tibiofibular synotosis or Kirschner wire group showed significantly better refracture-free survival than those with a pseudarthrotic fibula due to neglect or failed synostosis.There was significant difference between ankle joint fixed group and ankle joint unfixed group in mean refracture-free period.With a cross-sectional area median ratio of healing site of 0.15,refracture-free survival was better than those with a lesser value,and the difference was statistical significant.Cox proportional hazards regression analysis showed a negative correlation between the cross-sectional area and the refracture occurred ~=-1.989,P ＜ 0.05),fibular pseudarthrosis and refracture occurred were positively correlated ~=1.506,P ＜ 0.05),operative age and ankle joint status had no relationship with refracture occurred (P ＞ 0.05).Conclusion The major risk factors for refracture after union of congenital tibia pseudarthrosis with the use of intramedullary nail combined Ilizarov external fixator are the smaller cross-sectional area of pseudarthrosis and with fibula pseudarthrosis.
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Congenital pseudarthrosis of the tibia(CPT) is one of the most challenging diseases confronting pediatric orthopedic surgeon.The goal of the treatment is to achieve primary union and minimize the incidence of refracture and residual deformities.An optimal surgical treatment for the management of CPT is currently unavailable.Combined surgery can obtain good results.However, duo to heterogeneity of the disease, variable patients require individualized treatment to get a better prognosis.
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ObjectiveTo investigate changes of the level of serum thyroxine after different style operation on thyroid gland,for guiding replacement therapy after surgery. MethodsTo select patients randomly,include the excision of adenoma of thyroid、the lobectomy of unilateral thyroid、the subtotal ectomy of bilateral thyroid and total resection of bilateral thyroid quaque forty,among the total,the patients who suffererd the total resection of bilateral thyroid take thyroxine 40mg bid orally everyday. To detect the serum thyroxine in all of the patients, when preoperation、ten days after operation、one month after operation and three months after operation. ResultsThe serum thyroxine in patients who suffererd the excision of adenoma of thyroid were had no marked changes compare to preoperation; Serum FT3 、FT4 in patients suffer the lobectomy of unilateral thyroid are lower than preoperation when ten days after operation (P＜0.01) ;Serum FT3、FT4 in patients sufferred subtotal ectomy of bilateral thyroid were lower than preoperation,when ten days after operation and one month after operation( P ＜ 0.01 ), then returned to the level of preoperation when three months after operation;the serum thyroxine of patients who sufferred the total resection of bilateral thyroid had no marked changes compare to preoperation,when they taker thyroxine 40mg bid orally everyday. ConclusionThe serum thyroxine level of patients who suffer the lobectomy of unilateral thyroid、the subtotal ectomy of bilateral thyroid and total resection of bilateral thyroid is lower than preoperation at different degree, they should be given replacement therapy properly.
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[Objective]To study the results of Ilizarov technique in treatment of the severe rigid talipes equinovarus deformity.[Method]The Ilizarov technique was used in 9 patients(11 feet)with the severe rigid talipes equinovarus deformity,in which one patient with syringomyelia,and the orthers with congenital club foot.[Result]Prospective goals of correction were made in all the cases.The results were assessed by Garceau criterion.For 9 patients(11 feet),6 feets achieved excellent result,4 feets good and 1 foot poor results respectively.[Conclusion]Ilizarov technique in treatment of the severe rigid talipes equinovarus deformity achieves a good clinical result.
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[Objective]To observe radiographic morphology of femoral head at skeletal maturity of the Salter innominate osteotomy for treatment of patients with Legg-Perthes disease and evaluate sphericity of the femoral head and congruity of the hip.Based on system review of literatures,thes attempted to determine the indications and methods of treatment.[Method]Sixteen patients of Legg-Perthes disease were treated by Salter innominate osteotomy before 1994.The mean age of patients was six years and seven months at operation.Of them,only were 12 patients followed up 10 years and ten months avergely and mean ages of patients were 17 years and 2 months at last follow-up.Growth of plate of femoral head and Y cartilage of acetabulum was closed,indicating skeletal maturity.[Result]According to Stulberg classification of outcome,there were four cases in Stulberg class Ⅰ and six patient in class Ⅱ and one patient in class Ⅲ and one patients in class Ⅳ and no case in class Ⅴ.Lower limb of shortening was averagely 1.5 cm(ranged 0.8~2.0 cm)and the range of motion of hips was almost normal and no abnormal gait was observed.[Conclusion]The Salter innominate osteotomy could keep sphericity of the femoral head and congruity with acetabulum of the hip for children with Legg-Perthes disease and also could be in aid of preventing premature arthritis of hip.
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[Objective]To evaluate the results of intraoperative autologous blood transfusion in pediatric orthopaedic operation.[Method]Intraoperative blood salvage was reinfusioned in 17 patients by Cell Saver 5.The volume of reinfusion blood and complications were recorded.[Result]Among 17 patients,an average of(171?53)ml autologous blood was transfused intraopratively.The use of Cell Saver 5 was directly responsible for a 80% reduction in the total amunts of homologous blood.Four patients has experienced transient hemoglobinuria.No other complications appeared.[Conclusion]Reinfusion of blood salvage during pediatric orthopaedic operation is safe and effective.It is able to reduce the amount of homologous blood transfusion.