Osteosarcoma is a common malignant bone tumor in the skeletal system of minors .The five-year survival rate of patients with osteosarcoma is significanty improved by neoadjuvant chemotherapy combined with surgery.However,its mortality and morbidity rates remain quite high .With the development of molecular bi-ology and genetics ,gene therapy provides a new hope for patients with osteosarcoma .Researchers at home and a-broad have actively explored effective therapeutic targets .In this paper ,new progress in the study of gene -targe-ted therapy for osteosarcoma is reviewed .

AIM: To explore the change of the amount of GLUT4 protein at the plasma membrane of the rat skeletal muscle after high-fat feeding. METHODS: The animals were divided into three groups (ten for each): group I: control; group II: high-fat feeding; group III: high-fat feeding + dietary treatment. The rat model of insulin resistance (IR) was made by feeding high-fat diet for eight weeks. And then insulin-resistant rats were fed with chow diet for 4 weeks. Fasting plasma glucose and fasting serum insulin levels were measured before and after dietary treatment, respectively. Insulin treatment was achieved by intraperitoneal injection of insulin (10 unit insulin per kg body weight) 15 minutes before killing the animals. The right hindlimb skeletal muscle was rapidly dissected. Then the expression of GLUT4 protein at the plasma membrane in all the animals was assessed with Western bloting. RESULTS: The GLUT4 content at the plasma membrane in high-fat-fed rat skeletal muscle was significantly lower (about 31%) than that in controls (P

Objective To investigate the protective effect of desflurane on the brain against ischemia-reperfusion (I/R) injury and the underlying mechanism. Methods Ninety-six male Wistar rats weighing 250-300 g were randomly divided into 4 groups (n = 24 each) : group A sham operation; group B I/R; group C desflurane + I/R and group D 5-HD + desflurane + I/R. I/R was induced by occlusion of bilateral common carotid arteries combined with controlled hypotension for 10 min. In C group 1 MAC desflurane (5.9% ) was inhaled for 60 min before I/R. In group D 5-HD 5 mg?kg-1 was given i.v. before desflurane inhalation. The animals recovered from anesthesia at 30 min of reperfusion. The neurological behavior was evaluated by the clambering test, the overhanging test, the inclined plane test and the beam balance test. Animals were killed at 6, 24 and 48 h ( n = 8 each) of reperfusion in each group and the brains were removed for microscopic examination of area CA1 of hippocampus for the number of normal pyramidal neurons surviving I/R. Results Neurological behavior was greatly compromised by I/R at 6, 24 and 48 h of reperfusion. The animals behaved significantly better at 6,24 and 48h in C group but only at 6 h in D group than in B group. The number of normal pyramidal neurons in CA1 of hippocampus was significantly decreased by I/R at 6, 24 and 48 h of reperfusion. The number was significantly larger at 6, 24 and 48 h in C group but only at 6h in D group than in B group. Conclusion Desflurane preconditioning has protective effect on the brain against I/R injury. Activation of KATP channel is involved in the mechanism.

Objective To study the telomerase activity in the tissue adjacent to bladder cancer and to investigate its clinical significance. Methods Telomerase activity was detected with telomeric repeat amplification PCR (TRAP) assay. The telomerase activity in the tissue adjacent to bladder cancer was evaluated. Results Telomerase activity was positive in the tissue samples adjacent to bladder cancer in 10 of the 24 cases(42%). Telomerase activity in the adjacent tissue has been related with the tumour grades and stages. The tumour recurrence was also related with the telomerase activity in the adjacent tissue. Conclusions The detection of telomerase activity in the tissue adjacent to bladder cancer could be a prognostic marker for bladder tumor recurrence.

Objective To investigate the effects of aerosolized prostaglandin E1 (PGE1) inhalation on oxygenation and intrapulmonary shunt in acute lung injury (ALI) .Methods Eighteen healthy male pigs weighing 14-18 kg were anesthetized with intraperitoneal pentobarbital 50 mg?kg-1, intubated and mechanically ventilated (VT=10-15 ml?kg-1, RR= 16 bpm, FiO2=100%) . PaCO2 was maintained at 34-45 mm Hg. Anesthesia was maintained with intravenous infusion of ketamine-procaine-succinyl-choline. Swan-Ganz catheter was placed via right femoral vein. Right femoral artery was cannulated for BP monitoring and blood sampling. ALI was induced by intratracheal instillation of HCl (0.1 mol?L-1) until PaO2 was

Objective To explore SIRT4 gene expression in tumor tissue and investigate the clinicol-pathological features in osteosarcoma .Methods In this study ,SIRT4 protein expression was detected in 106 os-teosarcoma tissues and 36 paired neighboring non -tumorous tissues by immunohistochemistry and determined the correlation between the SIRT 4 expression and the clinicopathological features .Results SIRT4 protein was dra-matically decreased in osteosarcoma cells compared with neighboring non -tumorous bone cells .The low expres-sion of SIRT4 was notably associated with a poor overall survival and disease -free survival in osteosarcoma pa-tients.By using univariate and multivariate analyses ,we confirmed that the increased SIRT 4 expression was an in-dependent factor in predicting better prognosis for patients .Conclu is on SIRT4 expression might be an inde-pendent biomarker for prognostic evaluation of osteosarcoma .

Objective To investigate the role of glycogen synthase kinase -3beta(GSK3β)on invasion and metastasis of human osteosarcoma cells .Methods Expression and phosphorylation of GSK 3βwere examined in osteosarcoma cell lines and tumor tissue from osteosarcoma patients by Western blot .The effects of small mole-cule GSK3βinhibitors on cell metastasis were detected by cell invasion assay and cell migration assay .Results Osteosarcoma cell lines showed increased GSK 3βexpression and abnormal activity regulation .In tumor tissue of patients with osteosarcoma metastasis and non -metastasis,GSK3βwere detected expression and abnormal activi-ty,especially in tumor tissue of the patients with osteosarcoma metastasis .Inhibition of GSK3βactivity resulted in inhibiting cells invasion and migration in osteosarcoma cell line .Conclusion In this research,we demonstrated that GSK3βencouraged osteosarcoma cells metastasis .The result will open up a potential target for clinical treat-ment of osteosarcoma .

Objective To investigate the protective effects of lentivirus mediated Bcl-2-associated athanogene 1L (BAG-1L) over-expression on human neuroblastoma cells (SH-SYSY) induced by hypoxia/re-oxygenation,and to study its effect on the phosphoinositide 3 kinase serine/threonine protein kinase (PI3K/AKT) pathway.Methods SH-SYSY cells were cultured in vitro,and the cells at logarithmic phase were collected,and they were divided into recombined lentiviral infection group [infected by lentivirus containing BAG-1L and green fluorescent protein (GFP) gene],vector control group (infected by lentivirus containing GFP without BAG-1L gene) and cell control group (non-infection).Western Blot was used to detect the expression of BAG-1L in target cells after infection for 48 hours.SH-SY5Y cells were subjected to hypoxia for 8 hours and re-oxygenation for 24 hours,then the cell counting kit-8(CCK-8) was used to detect the cell activity,and the apoptosis was detected by flow cytometry after allophycocyanin labeled annexin V/7-amino actinomycin D (Annexin V-APC/7-AAD) staining.Western Blot was used to detect the protein expressions of BAG-1L,heat shock protein 70 (HSP70),AKT and phosphorylated AKT (p-AKT).Results After infection for 48 hours,exogenous BAG-1L protein bands were observed in recombined lentiviral infection group,but not observed in cell control group and vector control group.After hypoxia/re-oxygenation treatment,the cell viability in recombined lentiviral infection group was significantly higher than that in cell control group and vector control group (A value:0.689 ± 0.036 vs.0.425 ± 0.013,0.400 ± 0.012),apoptosis was significantly decreased [apoptosis rate:(26.97 ± 1.82)％ vs.(36.60± 1.45)％,(35.77 ± 3.74)％],the protein levels of BAG-1L,HSP70 and p-AKT were significantly increased [BAG-1L protein (gray value):2.405 ± 0.167 vs.0.529 ± 0.141,0.601 ± 0.099;HSP70protein (gray value):0.997±0.123 vs.0.634±0.091,0.584±0.106;p-AKT protein (gray value):1.234±0.118 vs.0.661 ± 0.210,0.712 ± 0.199,all P ＜ 0.01],but the protein level of AKT was slightly increased (gray value:1.103 ± 0.269vs.0.646 ± 0.188,0.791 ± 0.326) without statistically significant differences (both P ＞ 0.05).There was no significant difference in all parameters between cell control group and vector control group (all P ＞ 0.05).Conclusion Lentivirus mediated BAG-1L gene over-expression can protect nerve cells against hypoxic injury and apoptosis,and the protective effect may be related to the activation increase of pathway on PI3K/AKT.

Objective To investigate the protective effects of Bcl-2 associated with athanogene-1 (BAG-1) gene on human neuroblastoma cells (SH-SY5Y) injury induced by hypoxia/reoxygenation,and its influence on heat shock protein 70 (HSP70) expression.Methods The SH-SYSY cells at logarithmic growth phase were collected.Lenti virus mediated RNA interference (RNAi) technology was used to suppress the BAG-1 expression.The cells screened out can be divided into four groups:the cell control group with no lentivirus infection,lentivirus control group (containing only fluorescein protein lentivirus infection),BAG-1 siRNA group (BAG-1 siRNA silencing group),including BAG-1 siRNA-α group and BAG-1 siRNA-β group with lentivirus containing fluorescein protein (GFP) but at different BAG-1 siRNA target sites of silencing.Western Blot was used to detect the protein expression of BAG-1 and HSP70 in target cells after infectious recombination lentivirus for 72 hours;the Cell Counting Kit-8 (CCK-8) was used to detect the activity of four different group cells after hypoxia;the flow cytometry was used to detect the cell apoptosis;the HSP70 mRNA transcription level were detected by real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-qPCR) respectively in each group.Results After lentiviral infection for 72 hours,the Western Blot results showed that in the two BAG-1 siRNA silencing groups,the interference effect of BAG-1 siRNA-β group was superior to that of BAG-1 siRNA-α group.The cell viability of each group showed an increase followed by a decrease with the prolongation of hypoxia time,and reaching the peak at 8 hours.After hypoxia for 8 hours being given,the cell viability in BAG-1 siRNA-β group was significantly lower than that of the cell control group,lentivirus control group and BAG-1 siRNA-α group (A value:0.59 ±0.09 vs.0.94±0.12,0.90± 0.11,0.91± 0.14,P ＜ 0.01);the cell apoptosis rate was obviously higher in BAG-1 siRNA-β than that in the above three groups [(34.63 ± 3.46)％ vs.(14.83 ± 3.75)％,(19.93 ± 6.49)％,(16.40± 1.18)％,all P ＜ 0.01].There were no statistically significant differences in the HSP70 protein level and mRNA transcription level between BAG-1 siRNA-3 grroup,and the cell control group,lentivirus control group and BAG-1 siRNA-α group respectively (all P ＞ 0.05).Conclusion BAG-1 gene can play a role in protection of hypoxia nerve cells,reduce the apoptosis,and its protective effect can be independent of HSP70 gene.

BACKGROUND: Currently, the morphologic characteristics of the torn discoid lateral meniscus remain unclear, andmorphological indexes used to evaluate the discoid lateral meniscus tears still need to be explored. OBJECTIVE: To discuss the specificity and sensitivity of discoid lateral meniscal tears in children using morphological indexes. METHODS: Seventy-three patients (seventy-six knees) with torn discoid lateral meniscus underwentarthroscopy and MRI data of knee examinations were retrospectively reviewed. All MR images were double-blinded, independently, retrospectively analyzed by attending radiologist (doctor A) and sports physician (doctor B). The following morphological signs of discoid lateral meniscus, such as a internal displaced free edge sign, a hypertrophic free edge sign and a hypertrophied-horn sign, were selected to evaluate the discoid lateral meniscus tears. The likelihood of the discoid lateral meniscus tears was analyzed on basis of arthroscopic findings constituted the gold standard. Sensitivity, specificity and accuracy of MRI diagnosis of the discoid lateral meniscus tears by two physicians were calculated, and the consistency of diagnosis results by two physicians was evaluated using the Kappa statistics.RESULTS AND CONCLUSION: (1) The sensitivity, specificity and accuracy in the diagnosis of discoid lateral meniscustears by internal displaced free edge sign for doctor A were 19%, 94% and 53%, respectively, and for doctor B were 21%, 97% and 55%. (2) The sensitivity, specificity and accuracy in the diagnosis of discoid lateral meniscus tears by hypertrophic free edge sign for doctor A were 52%, 100% and 74%, respectively, and for doctor B were 57%, 97% and 75%. (3) The sensitivity, specificity and accuracy in the diagnosis of discoid lateral meniscus tears by hypertrophied-horn sign for doctor A were 26%, 97% and 58%, respectively, and for doctor B were 24%, 100% and 58%. (4) The sensitivity, specificity and accuracy in the diagnosis of discoid lateral meniscus tears by binding morphological index for doctor A were 86%, 91% and 87%, respectively, and for doctor B were 88%, 94% and 91%. (5) The Kappa statistics for discoid lateral meniscus tears respectively demonstrated a good inter-observer agreement (K > 0.75, P < 0.001). (6) These findings suggest that single morphological index used to evaluate the discoid lateral meniscus tears shows a good specificity, but which also shows a low sensitivity, while a binding morphological index can significantly improve the diagnostic sensitivity. Morphological characteristics may be reliable indexes to evaluate the discoid lateral meniscal tears in children.