Carotid endarterectomy (CEA) can effectively treat symptomatic carotid stenosis, however, perioperative stroke is the most important complication of CEA. The microemboli generated before and after the operation are the most important reason for causing perioperative stroke. In addition, the hypoperfusion and postoperative hyperperfusion syndrome caused by intraoperative clipping of carotid artery, as well as carotid restenosis or occlusion after CEA are all the causes of perioperative stroke. As a non-invasive, real-time monitoring tool, transcranial Doppler (TCD) can be widely used in intraoperative monitoring of CEA, It is able to detect a variety of perioperative blood flow changes and the production of microemboli, and thus effectively predicts the occurrence of perioperative stroke, and decreases the risks of perioperative stroke.

Objective To study the expression of immune and non-immune related cytokine in corneal grafts of chronic graft dysfunction after penetrating keratoplasty and reveal the possible mechanism of it.Design Clinical experimental study.Participants Including two groups.In chronic corneal allograft dysfunction(CCAD) group,corneas of 8 patients fulfilling the diagnostic criteria for CCAD who underwent repeated PK operation.Three normal corneas supplied by Shandong provincial eye bank of China were used as control group.Methods The conreal graft were analyzed by immunohistochemical studies.Moreover,the comprehensive analysis was carried out combining the patient's clinical data.Main Outcome Measures CD4~+,CD8~+,F4/80,TGF-?,bFGF and?-SMA in cornea. Results The results of immnunohistochemical studies showed that compared with normal corneas,full-thickness corneal grafts of eight corneas which underwent repeated PK operation had no CD4~+ and CD8~+ T lymphocyte infiltration.F4/80 positive inflammatory cells were more abundant near the stroma of five corneal grafts which underwent once PK operation.Full-thickness corneal grafts of other three corneas which underwent no immune rejection had no F4/80 positive inflammatory cells.The TGF-?positive inflammatory cells were more abundant near the epithelium and stroma of all corneal grafts.The moderate bFGF positive inflammatory cells were present in stroma of all corneal grafts.The?-SMA positive inflammatory cells were more abundant near stroma and the elastic layer of all corneal grafts.Conclusions After PK operation,the results of immnunohistochemical studies of the corneal graft which underwent CCAD showed that there is no evidence of clinical acute rejection.The abnormal expression of antigen presenting cells and non-immune cell specific factors suggest that immune or non-immune factors maybe involve in the occurrence and development of CCAD.

Objective: To study the chemical constituents of volatile oil from Radix Flemingiae philippinensis. Methods: The oil was obtained by steam distillation. The chemical compositions were separated and identified by GC/MS. The relative contents in the oil were determined by area normalization method. Results: Thirty-nine compounds were gotten from Radix Flemingiae philippinensis. The compounds of its relative content that was more than 4% were as follows: ?-himachalene (1), ?-himachalene (2), ?-himachalene (3), ?-guaiene (4), Italicene (5). Conclusion: Volatile oil by steam distillation from Radix Flemingiae Philippinensis was separated and ideutified up to 80 percent in component analysis.

The zinc finger antiviral protein (ZAP) specifically inhibits the replication of Moloney murine leukemia virus (MLV) and Sindbis virus (SIN), but has very modest inhibitory effect on HIV. Previous studies suggest that ZAP directly binds to the viral RNA and recruit the RNA degradation machinery to degrade the target RNA. The HIV-1 Tat and Rev are regulatory proteins which bind to HIV RNA. Tat and Rev were fused with ZAP in various manners. Two fusion proteins, ZAP-Tat and ZAP-Rev were found to be able to inhibit the expression HIV-1 vector.

Prosthesis for femoral head surface replacement was designed. Between January 2002 and December 2005,28 patients underwent femoral head surface replacement in Anyang People's Hospital. The diameter of the normal femoral head was determined by CT,and the prosthesis was selected according to the measured value ±1 mm. With the muscle gap approach,the center of the femoral head was determined according to the measured distance between the femoral head center and the culminated point of the round ligament fossa based on the mark of the culminated point of the round ligament pit. Additionally,the diameter of the femoral head was measured again to ensure the precise match of the prosthesis and acetabulum. X-ray films at 24-48 months show no loosening or dislocation of the prosthesis,which was well matched with acetabulum. Furthermore,the joint space did not remarkably change. The mean score was 63 before operation and 91.5 after operation respectively according to Harris sore. The successful rate is 92.86% (excellent in 23cases,good in 3,and improved in 2). The results show that femoral head surface replacement is an effective method for young patients with avascular necrosis of the femoral head at the stage of ARCO HI.

AIM: To screen the binding proteins to HMGB1 promoter by phage display technique. METHODS: HMGB1 promoter was incubated with phage display library. Unbound phages were eluted and phages bound to HMGB1 promoter were amplified. Twenty individual clones were randomly selected and identified by enzyme-linked immunosorbent assay (ELISA). Positive clones were characterized by DNA sequencing and the sequences were subjected for computer analysis. RESULTS: Positive phages binding to HMGB1 promoter were enriched after 4 rounds of biopanning. Twenty phage clones were selected and eleven clones of which were identified to bind specifically to HMGB1 promoter. The sequences in full length were obtained and searched for homologous sequences from GenBank. Altogether eight coding sequences were obtained, six of which were known proteins including activator protein-1(AP-1) and two of which were uncharacterized ones. CONCLUSION: Several proteins were obtained that bind specifically with HMGB1 promoter. The results will be useful for further studying the expression and regulation mechanism of HMGB1.

Objective To investigate the role of extracellular regulated protein kinase (ERK) signal pathway in mechanical stretch induced high mobility group box 1 protein (HMGB1) expression on alveolar epithelial cells (A549). MethodsA549 cells were cultured and seeded at 1×10~5 cells/ml in 6-well Bioflex cell culture plates. Subsequently, the cells were exposed to cyclic mechanical stretch at 14% (group B) elongation for 4 hours using Flexercell 4000T cell stretching unit. In group C, cells were pretreated with PD98059 for 2 hours before mechanical stretch. Cells in group A without stretch were served as control. The expression of HMGB1 protein and mRNA in A549 cells were detected by immunocytochemisty staining and RT-PCR, respectively. ERK activity was measured by Western blotting method. Results Immunocytochemisty staining indicated that the expression of HMGB1 protein in A549 cells was increased obviously in group B (P<0.05) and decreased in group C (P<0.05). Polymerase chain reaction (RT-PCR) showed that the expression of HMGB1 mRNA was also significantly increased in group B (P<0.05) and decreased in group C (P<0.05). Western blotting analysis confirmed the activation of ERK in A549 cells by mechanical stretch (P<0.05). PD98059, an inhibitor of ERK, might significantly inhibit mechanical stretch induced HMGB1 protein and mRNA expression in A549 cells (P<0.05). Conclusion Mechanical stretch could regulate the expression of HMGB1 gene and protein in A549 cells through ERK signal pathway.

Objective The intra-cranial arteries are more affected in Asian ischemic stroke patients. We thought that tandem or diffuse arterial occlusive disease may be more common among patients with extracranial internal carotid artery (ICAex) occlusive disease. In the current study, the frequency of other co-existing extracranial artery ( EA) or intracranial artery ( IA) stenosis in patients with ICAex occlusive disease and its risk factors were studied. Method One hundred and ninety-eight consecutive patients with ischemic stroke or TIA who were admitted to our hospital from Jan 2001 to May 2008 and who had stenosis (70% ) or occlusion of ICAex were enrolled in the study. All the patients were examined with carotid duplex and transcranial Doppler (TCD). Among them, 75 patients were also examined with DSA, 52 patients with intracranial MRA and 25 patients with extra- and intra-cranial CTA. Extracranial artery stenosis or occlusion were diagnosed with DSA/CTA in 93 patients, and with carotid duplex only in 105 patients. Intracranial artery stenosis was diagnosed with DSA/MRA/CTA in 118 patients, and with TCD only in 80 patients. Patients with presumably cardioembolism such as atrial fibrillation were excluded. Results Among the 198 patients, 172 (86. 8% ) had other combined EA or IA stenosis. One hundred and thirty-five patients (135/198, 68.2%) had other combined EA stenosis, which were more frequently found in the contralateral ICAex (71/198, 35.9%). One hundred and twenty-three patients (123/198, 62. 1%) had combined IA stenosis, which were more frequently found in MCA (75/198, 37.9%). 52 (52/198, 26. 3% ) had ipsilateral terminal ICA and/or middle cerebral artery stenosis. Conclusion Tandem or diffuse arterial occlusive disease is common among patients with steno-occlusion of ICAex . Therefore, it is important to evaluate other EA and IA stenosis before CEA or CAS in Chinese patients.

Objective To investigate the role of p38 MAPK pathway in the expression of high mobility group box 1 (HMGB1) in lung tissue in a rat model of ventilator-induced lung injury. Method Twenty-fonr healthy Sprague Dawley (SD) rats were randomly divided into 3 groups (n = 8 each) : group A, spontaneous breathing; group B, small tidal volume ventilation (Vt = 8 mL/kg) and group C, high tidal volume ventilation (Vt = 40 mL/kg). 1he animals in group B and C were mechanically ventilated for 4 hours and all animals were sacri-riced. The lungs were removed for: (1) lung lavage and determination of total protein contnt and WBC and neu-trophil counts in broncho-alveolar lavage fluid (BALF) ; (2) determination of W/D lung weight ratio and myelop-erexidnse (MPO) activity; (3) detennination of HMGB1 protein and mRNA expression and p38 MAPK activity in lung tissue. Differences within the groups were analyzed using One way ANOVA. Results The inflammatory re-sponse as evidenced by total protein (1.77 ± 0.68) g/L and WBC (106.55 ± 28.17) × 10~7/L in BALF, W/D lung weight ratio (7.16±1.02) and MPO activity (3.94±1.21) U/g were significantly higher in group C com-pared with group A (P <0.05); HMGB1 protein (0.64±0.17) and mRNA (1.17±0.45) expression and p38 activity (0.51±0.12) also significantly increased in group C (P <0.05). Of the above indexes, there were no statistical differences between group B and group A (P > 0.05). Conclusions High tidal volume ventilation in-daces acute lung injury, which may be related with upregulation of HMGB1 expression through p38 MAPK signal pathway.

Objective To construct the red fluorescent protein reporter gene vector containing high mobility group box 1 protein(HMGB1) promoter sequence and study the regulation mechanism of the expression of HMGB1gene under mechanical stretch.Methods HMGB1 promoter was subcloned into a red fluorescent protein vector,pDsRed1-1.After identified by PCR,enzyme digestion and DNA sequencing,the recombinant vector pDsRed1-1-HMGB1P was then transfected into HEK293 cells.Blank vector or pDsRed-1 was transfected into 293 cells and served as controls.The expression of red fluorescent protein and its reaction to mechanical stretch were observed under a fluorescent microscope.HEK293 cells transfected with pDsRed1-1 vector served as control.Results PCR,double restriction enzyme digestion and DNA sequence analysis showed that the recombinant vector,pDsRed1-1-HMGB1P,was constructed correctly.This vector was lowly expressed in HEK293 cells of resting state.But after stimulated by mechanical stretch,strong red fluorescence was observed.No red fluorescence was observed in the control cells.Conclusion A red fluorescent protein reporter gene vector containing HMGB1 promoter sequence has been constructed successfully and expressed highly in mammalian cells.Since it responds to mechanical stretch effectively,it can thus provide a convenient tool to study the regulation mechanism of the expression of HMGB1 gene by mechanical stress.