Objective To investigate the effect of rosiglitazone on the expression of platelet CD40 ligand (CD40L) in insulin-resistant rats, and to further determine the relationship between CD40L and insulin resistance. Methods 60 healthy male SD rats [(200±20)g] were randomly divided into 4 groups: control group (C), high fat group (HF), low dose rosiglitazone group (LR) and high dose rosiglitazone group (HR). Rats in group C were fed normal chow diet, and the others were given high fat chow diet. After 12 weeks, high dose of rosiglitazone (10mg/kg) was given to rats in group HR and low dose of rosiglitazone (5 mg/kg) was given to rats in group LR for 4 weeks. Rats in group HF and group C were given 0.9% sodium chloride solution. The level of sCD40L was measured by ELISA and the expression of platelet membrane CD40L was detected by immunoprecipitation and Western blot. The insulin resistance (IR) index was calculated by homeostasis model assessment (HOMA). Results HOMA-IR, sCD40L level and platelet membrane CD40L expression were higer in group HF than in group C (9.8±3.2 vs. 5.9±1.7, 367.3 ±35.3 vs. 232.3±120.6, 2.1±0.4 vs. 1.4±0.2, respectively, all P<0.05). Compared with the group HF, HOMA-IR, sCD40L level and platelet membrane CD40L expression were obviously decreased in group HR(5.4±1.1, 276.9±54.0, 1.4±0.3, respectively, all P<0.05), and there were no significant differences in HOMA-IR, sCD40L level and platelet membrane CD40L expression between group HF and group LR (P>0.05). Conclusions In insulin-resistant rats, the level of sCD40L and the expression of platelet membrane CD40L were higher. After treatment with high dose of rosiglitazone, sCD40L level and platelet membrane CD40L expression were decreased with the improvement of insulin resistance.

BACKGROUND:Acidic fibroblast growth factor (aFGF) possesses good effects on vascularization and osteogenesis.But whether aFGF can promote the vascularization in animals with early-stage avascular necrosis of the femoral head (ANFH) remains unclear.OBJECTIVE:To investigate the vascularization of aFGF composited by partially depreteinized bone (PDPB) in repair of early-stage ANFH.DESIGN,TIME AND SETTING:A randomized,controlled,animal experiment was performed in the College of Life Science,Nanhua University between January 2008 and January 2009.MATERIALS:Ribs from healthy,adult,New Zealand rabbits were prepared into PDPB by a series of physico-chernical methods including degreasing,deproteinization,partial decalcification and freeze drying,aFGF diluted with sterile distilled water was composited by PDPB particles to prepare artificial composite bone.METHODS:A bone window was made at the juncture of femoral head and femoral neck bilaterally in 24 healthy,adult,New Zealand rabbits.Rabbit models of bilateral ANFH were established by removing approximately 50% of cancellous bone and perfusion with 95% ethanol.Successful bilateral ANFH models were randomly divided into a blank group,a simple PDPB group,and an artificial composite bone group.PDPB and artificial composite bone were implanted into the PDPB and artificial composite bone group accordingly.The blank group did not receive any implantation.MAIN OUTCOME MEASURES:At 2,4,and 8 weeks after surgery,ink-injected specimens were prepared for microvessel count and microvessel area analysis.RESULTS:Microvessel number and microvessel area were least in the blank group,followed by simple PDPB group,and lastly the artificial composite bone group.There was significant difference in microvessel number and micrevessel area between artificial composite bone group and blank,simple PDPB groups (P＜0.05).CONCLUSION:Tissue-engineered artificial bone composited by aFGF and PDPB promotes vasculadzation in repair of earlv-staqe ANFH in rabbits.

BACKGROUND: Previous studies have demonstrated that acidic fibroblast growth factor (aFGF) combined with partially deproteinized bone (PDPB) (aFGF/PDPB) well promotes avascularization in animals with early-stage avascular necrosis of the femoral head (ANFH), but the histological results remain unknown.OBJECTIVE: To observe the histological repairing effects of aFGF/PDPB on early-stage ANFH in rabbits. METHODS: New Zealand rabbits were established models of bilateral ANFH and were randomly divided into a blank group, a simple PDPB group, and an aFGF/PDPB group. PDPB and aFGF/PDPB bone were implanted into the PDPB and aFGF/PDPB group accordingly. The blank group did not receive any implantation. At 2, 4, and 8 weeks after surgery, all animals were sacrificed for histological examination to observe the osteogenesis by hematoxylin-eosin staining.RESULTS AND CONCLUSION: Defects were filled with granulation tissues and fibrous connective tissues, only a little osteoid tissue formed at the borderline in the blank group at the end of the 8th week. In the PDPB group, a little new bone and cavitas medullaris formed. At 8 weeks, lots of graft was absorbed and cavitas medullaris formed with more osteoplasts and myeloid cells in it. The osteogenesis in the aFGF/PDPB group was better than that of PDPB group in each time point. At 4 weeks, the transplanted cavity was filled with osteoid tissues, a lot of osteogenic precursor cells and osteoblasts could be seen. Plenty of micrangium was observed, and osteoid tissues began to rebuild. At 8 weeks, the graft was replaced by bone tissues, and cavitas medullaris were formed with lots of bone marrow cells in it. At the borderline of the bone trabecula, there were lots of osteoplast and little osteoclasts, which may play a role in bone remodeling. There were mature bone cells in bone lacuna. Results indicate that aFGF/PDPB has better repair effect on rabbit model of ANFH than that of simple PDPB.

BACKGROUND: Tetramethylpyrazine has the protective effect against the central nervous system injury. The structural changes in Nissl bodies were regarded as a marker of neuron injury.OBJECTIVE: To investigate the effect of tetramethylpyrazine on the structure and the quantity of Nissl bodies of cerebral neurons in rat with epilepsy.DESIGN: A comparative study.SETTING: It was conducted at the Physiological Department of Medical School of Xianning College.MATERIALS: From September 2004 to March 2005, it was completed at the Anatomy Department of Tongji Medical College, Huazhong University of Science and Technology. Forty healthy SD rats aging 3-4 months, weighing (250±50) g and regardless of their gender, were selected.METHODS: Rats underwent anesthesia and craniotomy. Then their cerebral cortex were exposed for placing BL-410 Experimental System of Biological Function (TME, China) to record the bilateral EEG of the brain and the seizure in rats with penicillin-induced epilepsy group and the 10 mg/kg tetramethylpyrazine group, 20 mg/kg tetramethylpyrazine group and 40 mg/kg tetramethylpyrazine group. In control groups, the brains of rats were taken out at 1 hour after craniotomy. In penicillin-induced epilepsy group, their brains were taken out at 1 hour after penicillin-induced epilepsy. In 10 mg/kg tetramethylpyrazine group, 20 mg/kg tetramethylpyrazine group and 40 mg/kg tetramethylpyrazine group, after stable penicillin-induced epilepsy, tetramethylpyrazine was injected intraperitoneally at a dose of 10 mg/kg, 20 mg/kg and 40 mg/kg, respectively.When the greatest protective effect of tetramethylpyrazine appeared, the rats' brains were taken out. Brain sections were sliced. Nissl bodies were stained by thionine staining. Under light microscope, structures of Nissl bodies were observed and the images of Nissl bodies were quantitatively analyzed by HPIAS-1000 high acuity color pathologic diagram-writing analyzing system. In each group, the average absorbency of 15 fields was regarded as the average absorbency of Nissl bodies in that group.MAIN OUTCOME MEASURES: In all the groups, the structure and the quantity of Nissl bodies of cortical neurons in rats were studied.of the structure of Nissl bodies in external granular layer cells and external pyramidal layer cells. In control group, multi-layer blue-stained and clumplike or granule-like Nissl bodies could be observed. In penicillin-induced epilepsy group, Nissl bodies were completely resolved and disappeared. In 10 mg/kg tetramethylpyrazine group, Nissl bodies were partly or completely resolved. In 20 mg/kg tetramethylpyrazine group, the quantity of Nissl bodies was significantly increased as compared with those in penicillin-induced epilepsy group and the 10 mg/kg tetramethylpyrazine group. In 40 mg/kg tetramethylpyrazine group, the quantity and the structure of of the average absorbency of stained Nissl bodies in external granular layer cells and external pyramidal layer cells in rat cortex among all the groups.In penicillin-induced epilepsy group, the average absorbency were dramatically lower than that in control group (0.033±0.002, 0.756±0.035, t=4.93,P ＜ 0.01). In 20 mg/kg tetramethylpyrazine group and 40 mg/kg tetramethylpyrazine group, the average absorbency were significantly higher than that in penicillin-induced epilepsy group (0.435±0.011, 0.658±0.029, t=2.98,5.32, P ＜ 0.01). In 40 mg/kg tetramethylpyrazine group, the average absorbency were similar to that in control group (t=1.75, P ＞ 0.05).CONCLUSION: Tetramethylpyrazine can significantly elevated the concentration of Nissl bodies of neurons in rats with epilepsy. Changes in the structure and quantity of Nissl bodies of cerebral neurons may be closely associated with seizure, and tetramethylpyrazine can restore the structure and the quantity of Nissl bodies of neurons, regulate their functions and hereby, inhibit seizures.

Objective To assess the efficiency ,safety and feasibility of percutaneous renal sympa-thetic denervation (RSD) for elderly refractory hypertension patients .Methods Office and ambu-latory blood pressures ,serum levels of creatinine ,angiotensin Ⅱ and aldosterone ,estimated glo-merular filtration rate (eGFR) and rennin activity were measured in 20 elderly refractory hyper-tension patients before and 1 ,3 ,6 months after percutaneous RSD .Complications of percutaneous RSD were observed .Results The office and ambulatory blood pressures were 16 .9/11 .9 mm Hg (1 mm Hg=0 .133 kPa) ,24 .8/17 .1 mm Hg ,29 .1/20 .5 mmHg and 24 .2/17 .2 mm Hg lower 1 , 3 ,6 months after percutaneous RSD than before percutaneous RSD ( P< 0 .01 ) .No significant difference was found in serum creatinine level and eGFR before and after percutaneous RSD (P>0 .05) .The creatinine ,angiotensin Ⅱand aldosterone levels were significantly lower after percuta-neous RSD than before percutaneous RSD (P<0 .05) .Femoral artery hematoma was detected in 1 patient .Conclusion Percutaneous RSD is a safe ,effective and feasible procedure for elderly re-fractory hypertension patients .

BACKGROUND:Genetic modification by Shootin1 aims to effectively improve neural differentiation of bone marrow mesechymal stem cel s (BMSCs) in the injured spinal cord, thereby promoting functional recovery from spinal cord injury after cel transplantation. OBJECTIVE:To explore the nerve regeneration ability of transplanted BMSCs overexpressing Shootin1 in rats with spinal cord injury. METHODS:BMSCs were transfected using adenovirus-Shootin1 for 48 hours. Then, immunofluorescence staining was used to detect Nestin and NeuN expression levels in the transfected cel s under in vitro neuronal induction and differentiation. Animal models of spinal cord injury were made in rats using modified Al en’s method. Thirty minutes later, Shootin1-transfected BMSCs and non-transfected BMSCs were respectively injected into the subarachnoid space of the rats in the transfection and non-transfection groups, respectively. Rats in the model group were given no treatment. Five weeks after modeling, spinal cord samples were taken from each rat to make frozen sections for detection of nerve related markers RESULTS AND CONCLUSION:After 48-hour adenoviral transfection, Shootin1 expression was successful y detected in BMSCs. After 7-day in vitro induction, the cel morphology in the three groups varied, and there was no significant difference in the expression of Nestin and NeuN between the transfection and non-transfection groups. Basso, Beattie and Bresnahan scores were higher in the two cel transplantation groups than the model group. Increased expression levels of Nestin, NeuN, GFAP, MAP-2, ChAT and SYN were observed in both two cel transplantation groups, indicating a strengthened ability of nerve regeneration. Our experimental findings further confirm that BMSCs transplantation for spinal cord injury has achieved good outcomes, and Shootin1 protein plays a certain role in nerve regeneration and functional recovery after spinal cord injury. However, Shootin1 overexpression shows no obvious additional effects in combination with BMSCs transplantation, and further studies on the optimization of BMSCs transplantation for spinal cord injury are necessary.

Objective To study influence of nursing intervention on pain and satisfaction degree of pa-tients after lower limb fracture. Methods 200 patients of lower limb fracture were divided into the experi-mental group and the control group randomly with IOO patients in each group. Routine nursing was given to the contrtol group and special nursing intervention besides routine nursing to the experimental group. Pain and sat-isfaction degree was compared between the two groups and the results underwent χ2 test. Results The pain degree in the experimental group was much less than that of the control group, the satisfaction degree of the ex-perimental group was much higher than that of the control group. Conclusions Nursing intervention can re-liege pain degree of patients with lower limb fracture,make them go through postoperation pain stage and in-crease satisfaction degree with nursing.

Objective To study the nursing of bedsore prevention in patients with unstable pelvic fracture. Methods Bedsore forewarning risk assessment went on in 39 patients with unstable pelvic fracture by Braden scoring method. Results No bedsore was observed in these 39 patients with unstable pelvic fracture.The nursing result turned out to be satisfactory. Conclusions The incidence rate of bed-sore can be degraded in patients with unstable pelvic fracture by forewarning risk assessment with Braden scoring method and early nursing intervention.

Background:Pyruvate kinase M2( PKM2 ),which is highly expressed in cancer cells,plays an important role in cancer cell growth and aerobic glycolysis and is a promising target for cancer therapy. Aims:To investigate the effect of silencing PKM2 by siRNA on proliferation,apoptosis,migration,invasion and glycolysis of gastric cancer cell line SGC-7901 cells. Methods:SGC-7901 cells were divided into three groups:non-transfectd SGC-7901 cells,SGC-7901 cells transfected with empty plasmid and SCG-7901 cellls transfected with PKM2 siRNA. Expression of PKM2 was detected by fluorescent quantitative PCR and Western blotting at mRNA and protein levels. Intracellular expression and distribution of PKM2 was determined by immunofluorescence. CCK-8 assay,flow cytometry,migration and invasion experiment were used to assess cell proliferation,apoptosis,migration and invasion,respectively. Western blotting was used to determine the expression of glucose transporter-1( Glut-1)and lactate dehydrogenase A( LDHA). Spectrophotometry was used to detect levels of extracellular glucose and lactic acid and intracellular lactate dehydrogenase( LDH). Results:Compared with non-transfected group and negative control( empty plasmid )group,PKM2 mRNA and protein expressions were significantly decreased in PKM2 siRNA plasmid transfected group(P<0. 05). Intracellular expression of PKM2 was markedly decreased in PKM2 siRNA cells. After transfected with PKM2 siRNA plasmid,cell proliferation,migration and invasion were significantly inhibited,and cell apoptosis was increased significantly( P<0. 05 ). Expressions of Glut-1 and LDHA were down-regulated in PKM2 silenced cells( P <0. 05 ). Glucose uptake,lactic acid production and LDH activities were significantly decreased in PKM2 silenced cells(P<0. 05). Conclusions:RNA interference targeting PKM2 gene inhibits PKM2 expression in human gastric adenocarcinoma cell line SGC-7901 cells,thereby inhibits cell proliferation and glycolysis.

Objective To investigate the effects of ulinastatin pretreatment on oxidative stress response and hepatocyte growth factor (HGF) after hepatectomy in rats.Methods One hundred and twelve pathogen-free male Sprague-Dawley rats,aged 3 months,weighing 230-280 g,were randomly divided into 2 groups (n=56 each): group hepatectomy (group H) and group ulinastatin pretreatment (group U).Left and median lobe resection was performed and then liver ischemia was induced by blood flow occlusion of right and caudate lobes for 30 min,followed by perfusion in both groups.Ulinastatin 50 000 U/kg was injected intravenously at 5 min before occlusion in group U.Eight rats in each group were chosen and the blood samples were taken from the inferior vena cava for measurement of serum ALT and AST activities and HGF concentration before ischemia and at 1,6,12,24and 48 h of reperfusion.Then the right lobe were removed for determination of apoptosis,SOD and myeloperoxidase (MPO) activities,MDA content,expression of proliferating cell nuclear antigen (PCNA) and liver regeneration.Apoptotic index was calculated.Another 8 rats in each group were chosen and the 7 day survival rate was recorded.Results Compared with group H,the levels of ALT,AST,MPO aud MDA and apoptotic index were significantly decreased,and the levels of HGF and SOD,PCNA expression and liver regeneration were significantly increased at different time points in group U (P ＜ 0.05).There was no significant difference in 7 day survival rate between group H and group U (P＞ 0.05).Conclusion Ulinastatin pretreatment can strengthen liver regeneration after hepatectomy in rats,the underlying mechanism may be related to inhibition of oxidative stress response and increase in HGF production.