Objective To investigate the relationship among p38 mitogen-activated protein kinase (p38MAPK), NF-KB and monocyte chemoattractant protein-1 (MCP-1), and to study the role of p38MAPK, NF-κB and MCP-1 in diabetic nephropathy. Methods Protein expressions of p38MAPK and NF-κB, and mRNA expression of MCP-1 were initially investigated in rat mesangial cell line HBZY-1, which were incubated separately with 25mmol/L glucose, 100nmol/L insulin, 100 μmol/L H2 O2 and 100mg/L advanced glycosylation end products (AGEs). The relationship among p38MAPK, NF-κB and MCP-1 expression Was observed by usingSB203580, a specific inhibitor of p38MAPK. Results The expressions of p38MAPK, NF-κB and MCP-1 were increased in HBZY-1 cells incubated separately with 25mmol/L glucose, 100nmol/L insulin, 100μmol/L H2 O2and 100 mg/L AGEs. Expressions of NF-κB and MCP-1 were significantly reduced when p38MAPK was inhibited by SB203580. Conclusion p38MAPK, NF-κB and MCP-1 are involved in development of diabetic nephropathy, and p38MAPK stimulation is essential for the expressions of NF-κB and MCP-1.

The expression of cyclooxygenase-2 (COX-2) mRNA and protein was significantly increased in rat glomerular mesangial cells (RCMC) treated separately with high glucose, high insulin, H2 O2 and advanced glycosylation end products. The expressions of COX-2 mRNA and protein in RCMC induced by above 4 factors were distinctly inhibited by sodium selenite, suggesting that sodium selenite might play a significant role in the prevention of diabetic nephropathy via this mechanism.

Objective To determine the effect of Vibrio vulnificus cytolysin (VVC) inducing ap-optosis in human umbilical vein endothelial cell(HUVEC) and its possible mechanism. Methods The en-tire vvhA gene that encoding VVC from V. vulnificus strain GTC333 was amplified by PCR and sequenced af-ter T-A cloning. E. coli BL21DE3pET-42a-vvhA, a prokaryotic expression system of the vvhA gene, was then con-structed. Ni-NTA affinity chromatography was applied to purify the target recombinant protein rVVC, and SDS-PAGE plus Bio-Rad Agarose Image Analyzor were used to measure the output of rVVC and to determine the purity of rVVC extract. The activity of rVVC dissolving rabbit erythrocytes was detected by hemolysis test. DPNH chromotometry and TphBNa chromotometry were performed to examine the contents of LDH and K+ in the supernatants of rVVC-treated HUVEC cultures, respectively. The effect of rVVC inducing apepto-sis of HUVEC was detected by flow cytometry, rVVC was labeled with FITC and the location of FITC-labe-ling rVVC in HUVEC was observed by laser canfocal microscopy. Results The cloned whA gene had 96.09% and 98.26% similarities of nucleotide and amino acid sequences compared to the corresponding se-quences in GenBank. rVVC, with a dosage of 1 μg/ml, could dissolve rabbit erythrocytes (P<0.01). 10 μg/ml rVVC was able to promote the increases of K+ content (P<0.01) but no change of LDH content could be found in the cell supernatants. HUVEC was apoptotic after the cell was treated with 1～100 μg/ml of rVVC for 2 h. In the 5～240 min duration of co-incubation of FITC-labeling rVVC and HUVEC, the rV-VC gradually moved from surface to inner side of the membrane and then entered the cytoplasms. When FITC-labeling rVVC treated HUVEC for 30 min, most of the rVVC was found to be intracellular location. Conclusion rVVC has cytolytic activity. VVC has an ability to enter HUVEC and causes injury of HUVEC via inducing apoptosis, which may be the major pathogenic mechanism of VVC.

Absrtact: Osteoarthritis (OA) is a most common form of degenerative joint disease, primarily characterized by the degradation of articular cartilage, subchondral sclerosis and inflammation of the synovial membrane. Mesenchymal stem cells (MSCs), a multipotent adult stem cell population, can be isolated from many connective tissue lineages, including those of the diarthrodial joint. Joint-resident MSCs or MSC-like progenitor cells contribute to the maintenance of healthy microenvironment or to the response to trauma. The onset of degenerative changes in the joint related to abnormal condition or depletion of these endogenous MSCs and native host hyaline cartilage cells, leading to limited self-repair potential of the joint and advance of the degradation. To date, no acknowledged medical treatment strategies, including non-operative and classical surgical techniques, are efficient in restoring normal anatomy and function of hyaline cartilage in OA. This highlights an urgent need for better celled-based therapeutic strategies that supplement these functional cells exogenously to recover the tissue homeostasis and repair in joint cavity via chondrogenic and anti- in fl ammatory functions. In this review we focus on the role of native MSCs in healthy or OA joint and recent progress in cell-based researches utilizing culture-expanded chondrocytes, pluripotent stem cells, or MSCs from different sources for treating OA.

Objective To investigate the effect on electrical restitution of β1-adrenergic receptor antagonist esmolol administered during cardiopulmonary resuscitation in the porcine ventricular fibrillation model. Method Ventricular fibrillation untreated for four minutes was induced by dynamic steady state pacing protocol in 40 healthy male pigs, in which local unipolar electrograms were recorded using one 10-electrode catheter that was sutured to the left ventrieular epicarditan. During CPR, animals were randomized into two groups to receive saline as placebo or esmolol after two standards doses of epinephrine. At postresuscitation 2-hour, six pigs were randomly selected from each group and the second VF induction was performed. Local activation-recovery intervals (ARI) restitutions and the VF inducibility between control group and esmolol group were compared. Western blotting was performed to determine cardiac ryanodine receptor (RyR2) protein expression, and their phosphorylation status. Results No sig-nificant differences were observed at the restoration of spontaneous circulation between two groups. Higher postre-suseitation 2-hour survival rate was observed in the esmolol group. Esmolol significantly flattened ARI restitution slope, lessened regional difference of ARI restitution, decreased the VF inducibility, and alleviated RyR2 hyper-phosphorylation. Conclusions Esmolol given during CPR significantly improved postresuscitation 2-hour survival rate. Its effects on modulating electrical restitution property and intracellular calcium handling make up the most important reasons why β1-blockade significantly reduced the onset and maintenance of VF.

Objective To assess the effect of peroxisome proliferator-activated receptor γ (PPARγ) agonist on prostate epithelial cells in vitro.Methods The expression of peroxisome proliferator-activated receptor γ(PPARγ) was studied by immunocytochemistry and immunofluorescence study.The RWPE-1 human prostate epithelial cell line was treated with PPARγ agonist rosiglitazone 100 μmol/L for 48 h.Analysis of apoptosis was performed by Caspase 3/7 activity assay.Mitochondria depolarization was measured by using the potential-sensitive color,JC-1.The expression of apoptosis-related proteins-Bax was investigated by immunohistochemistry.Results PPARγ mainly located in nucleus and perinucleus.RWPE-1 cell line treated with PPARγ agonist rosiglitazone showed higher Caspase 3/7 activity (10636±1032 RLU) than in control (5936±620 RLU),P＜0.01 and significantly upregulated Bax level (8250±694 vs.6017±563)than in control group,P＜0.01.In addition,mitochondrial membrane potential was depolarized in rosiglitazone treated cells.Conclusions PPARmay play important roles in the pathophysiology of BPH.The mechanism might be that PPARγ regulates cell apoptosis.It is suggested that the mitochondrial and Bax pathway might be involved in signaling PPARγ induced cell apoptosis.

Objective To improve the accuracy of X-ray diagnosis of the deltoid ligament injury by evaluating the influence of plantar flexion on the diagnosis.Methods Twenty-four patients with ankle supination-extemal rotation fracture which was distal fibula fracture but no medial malleolus fracture were treated from February 2010 to December 2010.All the patients were taken Mortise radiograph with the ankle in four positions of plantar flexion(0°,15°,30° and 45°)applied with rotational stress and the deltoid ligament injury was identified with MRI.The medical clear space(MCS)and the superior clear space(SCS)on each radiograph were measured.The results were tested by a one-way analysis of variance model(ANOVA),based on the following criteria:(1)the MCS≥ 4 mm and MCS ＞ SCS,or when MCS≥5 mm and MCS ＞SCS.The MRI examination was taken as the gold standard to compare the influence of X-ray diagnosis of the deltoid ligament injury with different plantar flexions and different standards.Results The average MCS in four positions of plantar flexion(0°,15°,30°,and 45 °)with rotational stress were(4.10 ±0.79)mm,(4.55 ±0.72)mm,(4.99 ±0.56)mm and(5.71 ±0.86)ram,with statistical difference(P ＜0.05).The average SCS in four positions of plantar flexion(0°,15°,30°,and 45°)were(3.56 ±0.41)mm,(3.50 ±0.43)mm,(3.71 +0.44)mm and(3.93 ±0.51)mm,with no statistical difference in the four groups(P ＞0.05).With the MSC≥4 mm and MSC ＞ SCS as the diagnostic criteria,the prevalence of false-positive findings of deltoid injury based on the ratio with the four positions of plantar flexion(0°,15°,30° and 45°)were 50.0％,66.7％,88.9％ and 94.4％ respectively.With the MSC≥5 mm and MSC ＞SCS as the diagnostic criteria,the prevalence of false-positive findings of deltoid injury based on the ratio with the four positions of plantar flexion(0°,15°,30° and 45°)were 5.6％,11.1％,38.9％ and 77.8％ respectively.Condusions Different plantar flexion angle is an important factor for X-ray diagnosis of deltoid ligament injury of the ankle joint.The prevalence of false-positive findings of deltoid injury increases with the increase of plantar flexion.

Objective To determine the eugenol in Dijin spray by GC-MS.Methods The chromato-graphic column was DB-5 fused silica capillary column.Eugenol was determined by program temperature risen method:the initial temperature was 60 ℃ for 1 min,then raised to 200 ℃ at the rate of 10 ℃/min and kept for 5 min,and at last raised to 250 ℃ at 15 ℃/min and kept for 5 min.Electron energy was 70ev.The ions of 164,149,and 77 were selected for eugenol detection.Results The calibration curve of eugenol was linear in the range of0.116～1.16 mg/ml (r=0.999 4) with the average recovery of 97.84％ (RSD=0.47％).Conclusion The method was quick,accurate and repeatable.It was suitable for the quality control of Dijin spray.

Objective To investigate the value of pathological examination in the diag-nosis of lethal cases due to acute poisoning.Methods The macroscopic and microscopic find-ings in 28 autopsy cases (10 cases of toxic gas,7 cases of pesticide,6 cases of drug,3 cases of alcohol and 2 cases of chinese herbal medicine) died of acute poisoning during the period from October,2001 to June,2012 were retrospectively reviewed.Results Gross Changes were found on gross examination in 22 of the 28 cases studied.Of them,7 cases died of acute carbon mono-xide poisoning had showed cherry red of Shiban,muscle,visceral organs and blood.In addition,11 and 8 cases had been found gastric mucosa hemorrhage and epicardium petechial hemorrhage respectively.Histologically,myocardial interstitial hemorrhage was observed in 9 cases,pulmonary hemorrhage in 18 cases,bronchospasm in 12 cases,cerebral hemorrhage in 4 cases,hepato-cyte necrosis in 11 cases,renal tubular necrosis in 7 cases and gastric mucosa hemorrhage in 11 cases.In 6 cases of oral pesticides poisoning,besides they had all gastric mucosa hemorrhage,the change of character of gastric contents was found in 3 cases and pesticide odor from stomach in 3 cases.In addition,acute inflammatory and necrosis of gastric wall in 6 cases of oral pesticides poisoning were observed under the light microscope.Drug addicts often suffered from chronic lesions in multiple organs,such as chronic liver disease,fatty infiltration of the myocardium with myocardial atrophy,interstitial lung disease and pulmonary foreign body granuloma.Conclusions Pathology examination is helpful in the diagnosis of some lethal cases due to acute ooisoning,and it can provide pathological basis to study on treatment of acute poisoning.

Objective To compare the effect of application of autologous iliac alone and NovaBone combined with autologous iliac in the acetabular osteotomy of children with developmental dislocation of the hip (DDH).Methods Data of 113 cases of children with DDH who had undergone open reduction and acetabular osteotomy surgery from 2007 to 2011 were retrospectively analyzed.According to bone material using after acetabular osteotomy,the patients were divided into autogenous iliac bone graft group (52 cases,60 hips) and NovaBone combined with autologous iliac bone graft group (61 cases,67 hips).There were no statistical differences in gender,age,side,dislocation type,osteotomy and acetabular index between the two groups.The patients were evaluated by Lane's scoring criteria,Severin standards and McKay standards at 6 weeks,3 months,6 months,1-year and 2-year post-operation follow-up.The bone healing of acetabular osteotomy zone,radiography and function of hip were compared.Results 6 weeks and 3 months after operation,Lane bone healing score in NovaBone combined with autologous iliac bone graft group (6.4±1.3 points and 9.6±1.7 points respectively) was obviously superior to autogenous iliac bone graft group (4.7±1.5 points and 7.8±1.2 points respectively).And 6 months and 1 year after operation,the two groups were basically reached bone healing.The Severin standard results showed that the rate (94％,63/67) of Excellent and Good in NovaBone combined with autologous iliac bone graft group (excellent:41 hips,good:22 hips,Fair:4 hips) was significantly higher than the rate (83.3％,50/60) in autogenous iliac bone graft group (excellent:28 hips,good:22 hips,Fair:10 hips).The rate (16.7％,10/60) of Fair in autogenous iliac bone graft group was significantly higher than the rate (6.0％,4/67) in NovaBone combined with autologous iliac bone graft group.McKay standard results were consistent with the results of radiological evaluation.Conclusion As a novel bone defect repair material,NovaBone can promote early bone healing process of acetabular osteotomy areas.It also can improve the resistance of osteotomy area.NovaBone can get a satisfied result in acetabular osteotomy in children with DDH.