Objective Corneal endothelial decompensation is caused by many corneal diseases. It often results in severe clinical complications. Endothelial keratoplasty (EK) is a new therapy for corneal endothelial decompensation. This study aimed to investigate a new approach to establishing corneal endothelial decompensation animal model with Descemetorhexis technique in order to better understand the tissue response to EK. Methods Thirty New Zealand white rabbits were randomly divided into three groups according to different surgical procedures; corneal endothelial cells (CEC), Descemet's membrane and corneal endothelial cells (DM + CEC) as well as Descemet' s stripping with endothelial keratoplasty(DSEK) group and 10 eyes for each. The right eyes of rabbits were as surgery eyes. Other 10 rabbits were as DSEK donors. Corneal transparency, anterior chamber response and graft location were examined once per day for two weeks under the slit lamp. Comeal thickness was measured by ultrasound biomicroscope. Corneal endothelial cells were analyzed using vital staining with alizarin red and trypan blue in 2, 4 and 8 weeks after operation. Results The cornea in DM + CEC group remained opaque throughout the observation period. In CEC and DSEK group, corneal clarity was gradually restored and corneal thickness was significantly less than that in the DM + CEC group during the postoperative 8 weeks. There were significant differences in corneal thickness between the DM + CEC group and CEC group or DSEK group during the postoperative 8 weeks (P ＜0. 05). The vital staining showed that most Descemetorhexis area was not covered by endothelial cells even 2 months after surgery. Conclusion A new corneal endothelial decompensation model is successfully established for the study of corneal endothelial keratoplasty, which is helpful for understanding the wound-healing of rabbit corneal endothelium after Descemel' s membrane damage.

Objective p38 Mitogen-activated protein kinase (MAPK) is a crossing center of various pathways. In this study, protein transduction system based on human immunodeficiency virus (HIV)-1 transactivator of transcription (TAT), which is an efficient delivery peptide of the foreign proteins into cells, was employed to study p38 MAPK functions in eukaryotic cells. Methods p38 And its dominant negative form, p38AF, were constructed into pET-His-TAT vector correctly to verify that the recombinant plasmids were well-founded through restriction enzyme digestion and DNA sequencing. The two proteins, His-TAT-p38 and His-TAT-p38AF, were expressed and purified in Escherichia coli by SDS-PAGE. Then they were incubated with ECV304 cells respectively and readily transduced into cells in a time-dependent and dose-dependent manner. The cells were stimulated by sorbitol. Activating transcription factor (ATF) 2 phosphorylation level was checked using Western blot to assess the activity of endogenous p38. Results Compared with controls, it was found that His-TAT-p38 increased the level ofATF2 phosphorylation in sorbitol-stimulated ECV304 cells, while His-TAT-p38AF inhibited it, indicating p38 MAPK protein delivery system based on TAT was constructed successfully. TAT-p38 and its dominant negative form possessed high biological activity after transduction into ECV304 cells by TAT protein delivery system. The results showed that p38AF fused with TAT could inhibit the transduction of endogenous p38 signal pathway in part, and other pathway might regulate p38 phosphorylation. Conclusions Our study provides a novel pathway to inhibit p38 signal pathway and establish a new method to study p38 function.

BACKGROUND: Functional injury of brain and limb induced by hemorrhage of cerebral arteriovenous malformation could be restored by rehabilitative treatment OBJECTIVE: To investigate the curative effect of rehabilitation on restoration of the functional injury of the brain and limbs induced by bleeding of hemorrhage cerebral arteriovenous malformation (AVM) after embolization.DESIGN: Case analysis.SETTING: Department of Interventional Medicine of General Hospital of Chinese PLA.PARTICIPANTS: From January 1996 to December 2001, a total 56 patients who were diagaosed as cerebral hemorrhage or intraventricle hemorrhage by CT or MRI scanning was selected from General Hospital of Chinese PLA.METHODS: All 56 patients with cerebral arteriovenous malformation were diagnosed by DSA. Rehabilitative treatment was performed with α-butyl cyanoac-rylate or radian surgery. Rehabilitative therapy were as follow: [1] Electric stimulate: S J-Ⅱ type nerve-muscle therapy instrument was used to stimulate the muscles (flexor and extensor) of limbs in the involved side, intensity of which is from weak to strong and frequency of which was from 2 to 3 or 4 per day; [2] Acupuncture: The selected points are:Fengchi, Fengfu, Yintang, Shangxing, Baihui, Quchi, Waiguan, Hegu,Huantiao, Zusanli, Yanglingquan, Sanyinjiao, Taichong, et al. In those who suffered aphasia, lianquan, Jinjin, Yuye were selected. Once a day and a period of treatment was 2 weeks; [3] physical training: the passive movement on the joint in the involved limbs were performed in grade 0 to 1 of muscle strength, the balance training of sitting on bed or standing beside the bed in grade 1 to 2, walking training in grade 2 to 3. The training intensity should be from weak to strong gradually. The items of observation were the improvement in speaking, sense and activity in limbs in the involved side. Evaluation of myodynamia was referenced to 6 grading method of Lovett.MAIN OUTCOME MEASURES: [1] the degree of embolism; [2] the degree of speaking fluency, the sense of limbs and the strength of the muscles of the limbs.RESULTS: Totally 56 cases entered the final analysis without any loss. [1]Degree of embolism: Among 56 cases, 100% focus was eliminated for 77% in 43 cases, 90% was eliminated for 12% in 7 cases and 60%-80% was eliminated for 11% in 6 cases. [2] Fluency of language: Before the treatment of rehabilitation, they suffered different degree impediment of speaking in 14 cases, improvement in 14 cases and obvious improvement in 11 cases. [3] Sense of limb:The sense of limbs was improved in 23 cases and 18 of them improved obviously. [4] Strength of muscles: The strength of muscles in 0 to 1 grade in 3 cases, 1 to 2 grade in 11 cases, 2 to 3 grade in 15 cases. But after the treatment of rehabilitation, speaking ability was improved in 14 cases, and 11 of them improved obviously. The strength of muscles was. resumed to 2 or 3 grade from 0or 1 grade in 2 cases, and to 4 grade in 21 cases and 5 grade in 6 cases.CONCLUSION: Endovascular embolization is a minimal traumatic, the rehabilitative therapy which is secure and effective method for treatment of high risk cerebral AVM, can promote motion function, language function and ability of activity of daily living, and quality of life of patients with cerebral arteriovenous malformation after embolism.

Objective To investigate the clinical significance of brain-derived neurotrophic factor (BDNF) from peripheral serum in patients of vascular cognitive impairment (VCI).Methods Forty VCI subjects (including 10 mild cognitive impairment vascular(MCI-V) and 30 vascular dementia(VD)),and the control group for the same period in 40 healthy persons.Enzyme-linked immunosorbant assay (ELISA) was used to measure the serum levels of BDNF,statistical analysis was performed.Results The peripheral serum levels of BDNF in VCI (0.175 ±0.056) ng/L were lower than those of control group (0.211 ±0.061) ng/L,and there were significant differences (t =-2.752,P ＜ 0.05).The levels of BDNF showed no significant difference between MCI and VD ((0.195 ± 0.067) ng/L vs.(0.168 ± 0.052) ng/L,t =1.310,P ＞ 0.05).But they were both significantly lower than the control group (F =4.590,P =0.013).No significant differences were observed in the levels of BDNF between subcortical small vessel dementia (0.178 ± 0.057) ng/L and big vascular dementia (0.154 ± 0.042) ng/L (t =1.278,P =0.212).Conclusion BDNF participate in pathophysiology of VCI,and the serum levels of BDNF may be a candidate marker for clinical diagnosis of VCI.But serum levels of BDNF could not reflect the severity or the type of the VCI.

In order to investigate if there exists interaction between mitogen-activated protein kinase (MAPK) and signal transducer and activator of transcription 3 (STAT3) protein, and how the interaction regulates tumor necrosis factor-? (TNF-?) transcription activity, the human p38 and extracellular-signal regulated protein kinase 2 (ERK2) genes were amplified from human flag-p38 and flag-ERK2 by polymerase chain reaction (PCR) and cloned into pcDNA3-HA. Protein expression of the plasmids was examined by Western blotting. Co-immunoprecipitation was used to identify if there exists interaction between MAPK and STAT3 proteins. If the interaction was approved to be true, report gene system was applied to find how the interaction affect transcriptional expression of TNF-?. After STAT3 pathway was inhibited by RNA interfering, the action on TNF-? activity was determined. The results of DNA sequencing and enzyme digestion showed that the cloned p38 and ERK2 genes were correct, to be 1 080 bp or so. p38 and ERK2 proteins were expressed in 293T cell to be approximately 40 ku. Co-immunoprecipitation data showed that p38 and ERK2 proteins integrated with STAT3 protein in vivo. TNF-? reporter gene activity results found that protein complex of p38-STAT3 and ERK2-STAT3 coordinately increased TNF-? activity. After STAT3 was interfered, the TNF-? activity markedly decreased. These data indicated that there exists interaction between p38 and STAT3 protein, ERK2 and STAT3 protein. The complex of the proteins can coordinately regulate TNF-? expression. After interfereing STAT3 pathway, the coordinated action on TNF-? transcription activity might be obviously reduced.

Objective To evaluate the clinical,neuroimaging and electrophysiology features of 62 patients with multiple system atrophy(MSA).Methods Sixty-two cases with diagnosis of probable MSA were recruited in a retrospective studied.Clinical,neuroimage and external anal sphincter electromyography (EAS-EMG)data was retrospectively analyzed.Results In 62 cases(44 male and 18 female),the onset age was between 37 and 76.Among them,29 cases(46.8 % )were MSA-A,with orthostatie hypotension as the main clinical manifestation;24 cases(38.7 % )were MSA-C,with cerebellar ataxia ag the main chnical manifestation;9 cases(14.5 % )were MSA-P,with extrapyramidal symptoms as the main clinical manifestation.MRI showed that main lesion of MSA-A was in the cerebellum:that of MSA-C was in the cerebellum,pons and medulla;and that of MSA-P was in the putamen.Fifty-one cases did EAS-EMG and 46 cases showed neurogenie impairments.Nineteen cases were initially misdiagnosed with other diseases.Conclusions MSA is easy to be omitted or misdiagnosed at early stage.The diagnostie rate of MSA can be increased by the combination of clinical expressions,neuroimage,EAS-EMG and other necessary examinations.

Objective To compare the effects of different antiplatelet therapy on outcomes in patients with acute coronary syndrome (ACS). Methods 338 hospitalized patients with ACS were enrolled. They were assigned to three groups: group 1, aspirin alone after discharge, n=93; group 2, dual antiplatelet treatment of aspirin and clopidogrel after discharge for 6～12 months, then aspirin, n=127; and group 3, dual antiplatelet treatment of aspirin and clopidogrel after discharge for 2 years, n=118. All the patients were followed up for 2 years. The clinical data (basic clinical data, platelet count and serum lipids indeices), primary end point (cardiovascular death, nonfatal myocardial infarction and stroke) and hemorrhagic events (major hemorrhage, moderate hemorrhage and minor hemorrhage) within 1 and 2 years were analyzed. Results During 1 and 2 years, compared with group 1, the incidence of cardiovascular death and all primary end points of groups 2 and 3 decreased significantly (P＜0.05), but the nonfatal myocardial infarction and stroke did not （P＞0.05）. The difference was not statistically significant between groups 2 and 3 in all the end points （P＞0.05）. The difference of hemorrhagic events was not statistically significant among the 3 groups（P＞0.05）. Conclusion Dual antiplatelet treatment of clopidogrel plus aspirin for 2 years may decrease the mortality of cardiovascular disease while the incidence of severe hemorrhage doesn't increase.

Objective To investigate the special expression and diagnostic value of anti-mitochondrial antibody (AMA) in the muscle tissues of patients with mitochondria encephalopathy, lactic acidosis and stroke-like episodes (MELAS). Methods All cases including 10 cases of MELAS, which were diagnosed definitely by clinical, pathological and DNA analysis, and 2 cases of normal control and 3 cases of necrotic muscle fibers, underwent muscle biopsies in order to carry out AMA staining. At the same time the original MGT staining was observed. Results A large number of ragged brown fibers (RBF) were observed under light microscopy with AMA immunohistochemical staining in the muscle tissues of patients with MELAS. But no RBF were observed in control groups. Conclusion RBF might be the specific performance of paraplastic mitochondrial, which was similar to the ragged red fibers, so RBF might be a pathological index in evaluating abnormal mitochondrial function and in diagnosing mitochondriopathy.

OBJECTIVE: To evaluate the influence of fewer courses and prolonged intervals of chemotherapy on survival rate of advanced non small cell lung cancer (NSCLC) patients treated by sequential chemo-radiation therapy combined with traditional Chinese medicine (TCM). METHODS: From Jan. 2000 to Dec. 2001, 54 untreated advanced NSCLC patients (2 stage IIIa, 18 stage IIIb, 34 stage IV) were treated by sequential chemo-radiation therapy combined with TCM. The courses of chemotherapy were reduced and the intervals of chemotherapy were longer than that of the standard regimen. The efficacy and survival rate were documented and the prognostic factors were analyzed. RESULTS: Complete remission (CR) was observed in 1 case and partial remission (PR) in 20 cases. The overall objective response rate was 40.4%. Median survival was 15.3 months, 1-, 2- and 3-year survival rate were 53.7%, 28.9% and 9.6% respectively. The median survival of stage III and IV were 21.8 months and 12.5 months respectively. The 1-, 2-, and 3-year survival rates of stage III were 65.0%, 49.5%, 24.7% and that of stage IV were 47.0%, 23.3%, 0%, respectively. The quality of life was improved in most of the patients. Cox's proportional hazards regression showed that improved quality of life and treatment of TCM were the significant prognostic factors of overall survival. CONCLUSION: Chemotherapy and radiotherapy combined with TCM is beneficial to extending the interval of chemotherapy, improving the quality of life, and increasing the survival rate of advanced NSCLC patients.

Objective To study the protective effect of sufentanil preconditioning on hepatic ischemia-reperfusion injury in rats and to investigate the mechanisms whether may be by activating p38 MAPK signal pathway to promote p38 MAPK phosphoryla-tion .Methods Thirty SD rats(in either gender ,weighing 220-270 g) were randomly divided into five groups :Sham-operated group (Ⅰ) ,ischemia-reperfusion group(Ⅱ);sufentanil preconditioning group(5 μg/kg ,Ⅲ) ,SB203580(an inhibitor of p38 MAPK) group (Ⅳ) ,and dimethyl sulphoxide (DMSO) control group(Ⅴ) .Sample specimens were collected from each group at 240 minutes after reperfusion .Serum alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were measured by an automatic biochem-ical analyzer .Malondialdehyde(MDA) and superoxide dismutase(SOD) in liver tissue was measured .HE staining was used to ob-serve the hepatic pathological changes ,and to examine the expression of phosphor-p38 mitogen-activated protein kinases (p-p38 MAPK)of hepatic tissues by western blotting .Results Compared with group Ⅰ ,levels of AST ,ALT and MDA showed signifi-cantly increased in group Ⅱ-Ⅴ ,but levels of SOD decreased ,and obvious pathological changes were observed in the liver .In GroupⅢ significantly decreased the elevated levels of ASL ,ALT and MDA but increased levels of SOD ,and lessened hepatic pathological changes ,caused promoted p38 MAPK phosphorylation at 240 minutes after reperfusion .The protective effects of sufentanil precon-ditioning were abolished by SB203580 pretreatment .There were no significant differences between group Ⅴ and group Ⅱ .Conclu-sion Sufentanil preconditioning can reduce the hepatic ischemia-reperfusion injury .The protective mechanisms may be by activating p38 MAPK signal pathway to promote p38 MAPK phosphorylation .