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

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 .

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 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 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

AIM:To observe the effects of normal mesenteric lymph (NML) on the lung, heart and liver inju-ries and the phosphorylation levels of p 38 mitogen-activated protein kinase ( MAPK) , extracellular signal-regulated kinase (ERK) 1/2 and c-Jun N-terminal kinase (JNK) in the mice with endotoxic shock (ES).METHODS: The NML was drained form health male BALB/c mice for the intervention of ES after the removal of cellular constituent .Lipopolysaccha-ride (LPS, 35 mg/kg) was intraperitoneally injected into the mice for the establishment of ES model .After 60 min of LPS injection, the administration of NML (1/15 of whole blood volume) was performed through the femoral artery in NML +ES group.Meanwhile, the mean arterial pressure (MAP) was monitored during the experiment .At 6 h after intraperitoneal in-jection of LPS or the corresponding time point , blood samples were harvested from the heart through apical centesis for de-termination of the biochemical indexes to reflect myocardial and hepatocyte injuries .Simultaneously , the lung , heart and liver tissue specimens from a fixed location were harvested for the observation of histomorphology and the measurement of phosphorylation levels of p38 MAPK, ERK1/2 and JNK.RESULTS:Compared with sham shock (SS) group, MAP in ES group and NML+ES group remarkably decreased at multiple time points after intraperitoneal injection of LPS .However, MAP in NML+ES group at 80 min, 90 min, 190 min, 210 min, 240 min, 250 min, 340 min, 350 min, and 360 min were significantly increased compared with ES group .There were normal structures in the lung , liver and myocardium of the mice in SS group, while the morphological damages of these tissues appeared in ES group .Meanwhile, the damages were attenuated in the mice of NML +ES group.The activities of AST , ALT and CK-MB in the plasma in ES group were remark-ably higher than those in SS group .The CK-MB activity in NML+ES group was also increased compared with SS group , and the activities of AST and LDH-1 were lower than those in ES group .At 6 h after LPS injection , the phosphorylation levels of p38 MAPK, ERK1/2 and JNK in the lung tissues were remarkably increased .Meanwhile , no statistical difference of these indexes between the myocardial and hepatic tissues was observed .NML intervention decreased the phosphorylation levels of p38 MAPK in the lung tissues , and p38 MAPK, ERK1/2 and JNK in the myocardial tissues .CONCLUSION:The NML administration alleviates multi-organ injuries and reduces the phosphorylation level of p 38 MAPK in the lung tis-sues in the mice subjected to ES .

Objective: To explore the effects of different proportions of Fructus Cnidii (Shechuangzi) and Psoralea corylifolia (Buguzhi) on highly metastatic human breast cancer cell line MDA-MB-231BO and bone marrow stromal cell line ST-2 in vitro. Methods: Thirty-six female SD rats were randomly divided into 6 groups to prepare the drug-medicated sera by administering with different proportions of Fructus Cnidii and Psoralea corylifolia, including 4:0 group, 3:1 group, 1:1 group, 1:3 group, 0:4 group and control group. MDA-MB-231BO cells and ST-2 cells were cultured in Dulbecco's modified Eagle's medium containing drug-medicated serum. Inhibition rates of MDA-MB-231BO cells and ST-2 cells were measured by methyl thiazolyl tetrazolium (MTT) method; migration ability of MDA-MB-231BO cells was tested by a cell migration experiment; alkaline phosphatase activity (ALP) of ST-2 cells was measured by using 4-nitrophenyl phosphate disodium salt, and mineralized nodule formation of ST-2 cells was measured by alizarin red staining. Results: Sera contaning different proportions of Fructus Cnidii and Psoralea corylifolia inhibited the migration activity of MDA-MB-231BO cells as compared with the blank serum, and serum contaning Fructus Cnidii and Psoralea Corylifolia at proportion of 1:1 had the best function (P<0.01). Fructus Cnidii and Psoralea corylifolia at ratio of 1:1 also enhanced the ALP activity of ST2 cells (P<0.05) and increased the number of mineralized nodules of ST2 cells (P<0.01). Conclusion: Kidney-warming recipe of Fructus Cnidii and Psoralea corylifolia can inhibit proliferation and migration of MDA-MB-231BO cells and increase the activity of ST-2 cells.

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.

Objective To investigate the effects of desflurane on the delayed rectifier potassium current (Ik ) in acutely dissociated rat parietal cortical neurons. Methods Wistar rats between 10- and 14-day old of both sexes were used. The parietal cortical neurons were acutely dissociated enzymatically. The extracellular fluid saturated with 0.3,0.6 and 0.9 mmol/L desflurane was added to the culture dish, then the effects of different concentrations of desflurane on Ik were investigated by using the whole-cell patch-clamp technique in acutely dissociated rat parietal cortical neurons. Results IK was inhibited by desflurane in a concentration-dependent manner ( P ＜0.01). The V1/2 of the activation and inactivation curves and the slop factor had no change after giving 0.6 mmol/L desflurane (P ＞ 0.05). Conclusion Desflurane inhibits delayed rectifier potassium channels of parietal cortical neurons of rats in a concentration-dependent manner, and has no effect on the activation and inactivation of delayed rectifier potassium channels, indicating that the change in the excitability of the channel is not involved in the mechanism of inhibitory effect of desflurane, and the other reasons may be involved in the mechanism.

Objective To investigate the toxic reaction,toxic organs or target tissues of protamine recombinant human insulin (Insulin NPH),and provide basis for clinical trials by single dose toxicity test in mice,repeated toxicity and immunogenicity of Beagle's dogs,and systemic active allergy in guinea pig.Methods ① Using maximum dose method,mice in single dose toxicity test were sc injected with normal saline (NS),vehicle,and Insulin NPH (2092-2488 IU/kg),the toxic reactions after injection were monitored.② In repeated toxicity study,Beagle's dogs were sc administrated with vehicle,the original (Humulin NPH,1.5 IU/kg)and different doses of Insulin NPH (0.5,1.0 and 1.5 IU/kg) for 30 d continuously,followed by a 14-d recovery.During the administration and recovery period,general observation,local irritation,body weight,anus temperature,blood glucose,and electrocardiogram (ECG) were checked,moreover,hematology,serum biochemistry and urine were detected.Also,organic weights and histopathological examination were conducted.Binding antibodies in dog serum were measured by indirect ELISA method in immunogenicity test.③ In systemic active allergy study,cavies were sc injected with low-and high-dose (4 and 12 IU/kg) Insulin NPH,normal saline and vehicle.Besides,ova as positive control was also included.After five times of sensitization test with above doses,the excitation reactions of iv injection with tripled sensitizing doses were observed.Results No obvious toxicity was observed in mice after injected with 165 times of usual clinical dose of Insulin NPH.Repeated toxicity study of Beagle's dogs revealed that 1.0 IU/kg was the no-toxic-effect dose (NOAEL) for Insulin NPH,which was equivalent to 2 times of clinical dose.No bindingantibodies were found in immunogenicity test.There was no obvious allergic symptom in the active systemic allergy study of guinea pig.Conclusion Under the experimental conditions,no serious toxicity of Insulin NPH is found.