AIM: To investigate the expressions of the cyclin dependent kinase inhibitor (CKI) in human corneal epithelium.METHODS: The expressions of CKI, P27, P21 and proliferating cell nuclear antigen (PCNA) were tested in different regions of corneal epithelium by SP immunohistochemistry.RESULTS: Limbal basal cells stained positively for PCNA while central corneal epithelium cells stained negatively for PCNA, their difference was statistically significant (P＜0.01).Positive staining for P27 and P21 were observed in central epithelium, but there was no positive staining in limbal epithelium. Their difference were also statistically significant (P＜0.01).CONCLUSION: The different expressions of CKI P27, P21and PCNA in different corneal epithelial regions suggest that in limbal basal layer there are a group of cells that have higher proliferative capacity staying in G1 status, namely stem cell.

Objective To investigate genotypes and antimicrobial susceptibility of Rhizobium radiobacter (R.radiobact-er)isolated from pediatric patients with bacteremia.Methods R.radiobacter strains from blood cultures of pediatric pa-tients with bacteremia from February 2013 through February 2014 were collected.16S rDNA fragment was amplified by polymerase chain reaction(PCR),bacterial genotypes were identified by pulsed-field gel electrophoresis (PFGE),antimi-crobial susceptibility testing were performed by Kirby-Bauer disk diffusion method.Results 13 isolates of R.radiobacter were isolated from 12 children with positive blood culture,which including 3 genotypes,the homology of 16S rDNA fragment and R.radiobacter IFM 10623 was up to 99%.According to CLSI 2003 standard,these 13 R.radiobacter were resistant to ceftazidime,piperacillin,ticarcillin,piperacillin/tazobactam,ticarcillin/clavulanic acid,aztreo-nam and polymyxin B,but sensitive to cefepime,doripenem,imipenem,meropenem,tobramycin,netilmicin,cip-rofloxacin,levofloxacin,and norfloxacin.In addition,cefotaxime and ceftriaxone showed larger inhibition zone. Conclusion Therapy of R.radiobacter infection should be based on antimicrobial susceptibility testing results;cefo-taxime and ceftriaxone are better selection for the therapy of R.radiobacter infection in pediatric patients.

[Objective]To investigate the efficacy of tissue-engineerring artificial bone in treating rabbit bone defect.[Method]Rabbit bone marrow stromal cell were separated for cell culture,and subculture was done to increase the number of bone marrow stromal cell.A combined grafting material was made of bone marrow stromal cell and bioglass.The mixture,ilium and bioglass were used respectively to repair segment defects created in the radius of rabbits.The repair capability was assessed by figuration,radiography,histochemistry stains,scanning electron microscopy and biomechanical analysis at 12 weeks after operation.[Result]The amount of new bone formation with tissue-engineering artificial bone was more than that of other 3 groups.At 12 weeks the defect was bridged with the appearance of marrow cavities,but the bony healing was not found in the blank controls,which were repaired by fibrous tissue.After 12 weeks,the new bone formation in the group of the combined bone marrow stromal cells and bioglass showed the best performance.There was significant difference between groups.[Conclusion]Bone marrow stromal cell is a kind of good bony stem cell.The mechanism of osteogenesis may attribute to both intramembranous and endochondral ossification.The tissue engineering structuring bone marrow stromal cell is a promising method to repair bone defects.

Objective To investigate the effect of chronic stress on the expression of uncoupling protein 4 (UCP4) and Bcl-2 protein in hippocampal neurons of rat depression model.Methods Rat depression models were established by chronic unpredicted mild stress.All rats were randomly assigned to 2 groups:control group and model group.Flow cytometry was applied to detect the apoptosis rate and mitochondrial membrane potential.The level of LDH was measured by enzymes labelling instrument.The number of neurons was measured by immunohistochemistry.The expression of UCP4 and Bcl-2 protein was measured by Western blotting.Results After chronic stress,the apoptosis rate((4.35±0.19) ％)and LDH activity ((445.50±91.70) U/mg) in hippocampal tissue in the model group was significantly higher than the control group((0.34±0.06) ％,(167.20±63.40)U/mg).Compared to control group,the number of hippocampal neurons ((72.50±4.25) vs (45.30±2.54)) and the mitochon drial membrane potential decreased in the model group.The expressions of UCP4 and Bcl-2 protein in hippocampal tissue were significantly lower than the control group.Conclusion Chronic unpredicted mild stress can lead to apoptosis in rat hippocampal neurons,which is related with decline of mitochondrial membrane potential and low expression of UCP4 and Bcl-2 protein.

Objective To explore the effects of specific T-cell immunity against prostate cancer(PC) cells induced by dendritic cells(DCs) derived from fetal organs.Methods Mononuclear cells(MNCs) were obtained from the fetal bone marrow and liver.Then MNCs were cultured in medium with induction of rhGM-CSF,rhIL-4 and rhTNF-?to get DCs.Lysates of DU145 containing HSP-peptide complex were prepared by 50%-70%(NH4)2SO4 saturation.T lymphocytes from fetal spleen were co-cultured with DC loading DU145 antigen for 72 h,whereby CTL was obtained.The cytotoxicity of CTL against DU145,PC3 and EJ was detected by MTT assay.Results Mature DCs were induced from fetal organs,which expressed CD1a,CD_(86),HLA-DR and CD_(83) at high levels.DC stimulated with tumor lysates transformed T cells to specific CD~+_8 CTL.Phenotype of CD~+_8 cell was(14.09?(2.46))% before transformation,and(62.76?2.64)% after transformation,respectively(P

Objective To explore the influence of acute myocardial infarction (AMI) patients with coronary lesions to the ST segment resolution (STR) and clinical prognosis after percutaneous coronary intervention (PCI).Methods 380 patients with AMI were detected twice each hour for the situation of STR,before and after coronary angiography and PCI.According to the STR,the patients were divided into STR (≥ 50％) group and no STR (＜ 50％) group.The relationship between STR and clinical prognosis of hospitalization or follow up were analyzed.Results The STR rate of patients with coronary multivessel disease or proximal lesions after PCI was lower than the rate of patients with coronary single vessel disease or distal lesions (P ＜ 0.05).The incidence rate of major cardiac events and LVEDd during hospitalization in patients with proximal coronary lesions was obviously higher than patients with distal lesions,while LVEF was lower than distal lesions(P ＜0.01).It had no significant differences in LVEF,LVEDd between coronary multivessl disease and single vessel disease during follow up (P ＜ 0.01).Conclusion It has significant relationship between coronary multivessel disease,proximal coronary,STR resolution in patients with AMI after PCI and the prognosis during hospitalization and follow up.

Objective To explore the relationship between ST-segment level,T-wave early inversion and patients' left ventricular function,prognosis specifically for ST-segment elevation acute myocardial infarction patients,which underwent emergency PCI.Methods 62 patients (M:40,F:22 ) with acute myocardial infarction were randomly divided into groups as follows:according to whether one hour ST segment's changes more than 50％ after PCI,they were divided into group A and B:group A (PCI after one hour ST segment ≥50％ ),group B (PCI ST segment after one hour ＜50％ ).According to whether 24-hour T wave inversion after PCI,they were divided into group C and D:group C (24-hour T-wave inversion),group D (24 hours without T wave inversion).After six months,the different LVEF betweeen group A and B,C and D was observed.Results Of group A and group C,after 6 months,the left ventricular function recovery was better than that of group B and group D ( P ＜ 0.05 ).Conclusion The analysis of ST segment and T wave are simple,practical and effective methods to predict myocardial reperfusion,especially for PCI therapy in patients with acute myocardial infarction.

Objective To investigate the changes of serum bilirubin level in elderly patients with acute cerebral infarction and its significance.Methods 164 hospitalized elderly patients,who suffered from acute cerebral infarction within 1 week after onset,were divided into 2 groups according to age:group A aged over 60 years(n=85) and group B aged 40-60 years(n=79),and 66 healthy subjects aged over 40 years were collected as controls(group C).Serum bilirubin levels in all subjects were determined.The ratio of pulse pressure over mean arterial pressure(PP/MAP) in group A and B was calculated.Nerve function scores in the three groups were detected before and after 2 weeks of treatment.Meanwhile,the data of risk factors including blood glucose,blood pressure,blood lipids,smoking and drinking in group A and B were collected.Results Compared with group C,serum total,direct,indirect bilirubin levels were increased in group A and B(both P＜0.01),and the change was smaller in group A than in group B(P＜0.05).The nerve function scores was lower in group A than in group B before and after treatment [(35.2±12.6) vs.(44.3±7.9),(40.7±9.1) vs.(51.3± 4.1),t=5.58,9.73,both P＜0.01],but PP/MAP and the numbers of risk factors were higher in group A than in group B [(0.46±0.06) vs.(0.38±0.06),93.01 vs.71.20,both P＜0.01].There were no significant correlations of serum total,direct and indirect bilirubin levels with nerve function scores in group A or B(all P＞0.05).Conclusions Serum bilirubin level is increased in patients with acute cerebral infarction,but the endogenous antioxidant capacity is decreased because of aging,multiple risk factors and more serious atherosclerosis in elderly patients,and the increment of bilirubin level is relatively smaller in acute cerebral ischemia,leading to the reduced protective effect against stress.Serum bilirubin level may influence the prognosis in patients with acute cerebral infarction.

Objective To analyze the mechanism of fentanyl combines midazolam,by comparison with some of the commonly used anesthetic method to determine in the ethical advantages of anesthesia.Methods 45 rats were randomly divided into three groups,Group A:1％ anesthetized by intraperitoneal injection of sodium pentobarbital 40 mg/kg; Group B:5 ％ ketamine 120 mg/kg intraperitoneal injection; Group C:midazolam 5 mg/kg + fentanyl 0.05 mg/kg Last intravenous anesthesia.Observe the effects of anesthesia.Results The rats which are used of fentanyl conbines midazolam anesthesia,the onset of anesthesia time:0.02 ± 0.03 min,maintenance of anesthesia time:89.73 ± 22.59 min,the the narcotic operation time:1.51 ± 0.30 min,the success of anesthesia cases:15 cases.Conclusions Fentanyl combines midazolam compared with the currently used method of anesthesia,the advantages of the more prominent,it has significant improvement and protection about the security,utilization and recovery ethics in experimental rats.

ObjectiveTo investigate effects of combined regimen of low dose gossypol with steroid hormones on apoptosis of spermatocyte and phagocytosis of sertoli cells. Methods Adult male rats were divided into four groups randomly, group GH: rats were fed orally with gossypol(GA) [ 12.5mg / (kg·d) ] and desogestrel(DSG)[125μg/(kg·d) ]/ethinyloestradil(E)[25μg/ (kg·d) ]/testosterone undecanoate(TU)[100mg/(kg·d) ]; group G: a single does of GA[12.5mg/(kg·d ) ] was given; group H: DSG[125μg/(kg·d) ]/E[25μg/(kg·d) ]/TU[100mg/(kg·d) ] were administered; group C: rats only received vehicle(1% methyl cellulose). Testes from all the rats were removed at 4, 6 and 8 weeks after treatment for counting of spermatocyte and round spermatid using stereological assay, and for detecting apoptosis of spermatocyte and phagocytosis of sertoli cell by TUNEL and oil red O staining respectively. Results In GH group, the number of spermatocyte and round spermatid were reduced, while apoptosis of spermatocyte and staining of oil red O in seminiferous epithelium increased significantly. All changes were caused by steroid hormones in the combined regimen. Conclusion Induction of apoptosis of spermatocyte and then being phagocytosed by Sertoli cell is one of antifertility mechanisms of the combined regimen.