BACKGROUND: Techniques in the focal cerebral ischemia model in mice have been well developed in some foreign countries. However, rats are commonly used in cerebral ischemia studies due to resource availability in China.OBJECTIVE: To establish reliable and reproducible focal cerebral ischemia reperfusion model in mouse to explore the molecular mechanism of cerebral ischemia reperfusion process in genetic level.DESIGN: Randomized controlled study.SETTING: Fundamental Medical Research Institute of Chengde Medical College. MATERIALS: The experiment was carried out in Fundamental Medical Research Institute of Chengde Medical College from September 2002 to May 2003. Totally 30 Kunming mice (25 to 30 g), provided by Experimental Animal Center, Chengde Medical College, were randomly grouped as follows: control group, 6, 3-hour occlusion and 18-hour, 24-hour reperfusion, with 6 mice in each group respectively.INTERVENTIONS: Nylon thread (qb0.128 mm) was soaked in paraffin wax and inserted into internal carotid artery in mouse to reversibly occlude the middle cerebral at the right side after 3 hours, then was removed, and reperfusion was performed in 18, 24 hours, respectively.MAIN OUTCOME MEASURES: The infarct volume was observed and neurological deficit scored was analysis.of mice in operation group was shown as head turning to left, adduction of left forelimb, internal rotation of shoulder, decrease of muscular tension of left forelimb, unable to straighten completely during tail lifted. At independent activity, mice turned to left as the left hidlegs as the center of a circle; under the abdomen, left forelimb was crossed with right forelimb as scissor shape. Palsy of left limbs was observed obviously, especially forelimb.tion was shown clearly with TTC. The infarct volumes were (13±2.3) mm3 and (16±4.5) mm3 for 18 hours and 24 hours reperfusion and 3-hour ischemia, respectively. Pathological changes were observed under light microscope after ischemia. The infarct volume increased as the increasing of reperfusion time, including 3-hour blocking and 18-hour and 24-hour reperfusion.CONCLUSION: Using suture method to establish focal cerebral ischemia reperfusion model in mice, it is less invasion and trauma without requiring skull opening. Ischemia position is relatively fixed, thus, ischemia reperfusion time can be controlled precisely. It is also an ideal animal model to study pathological changes in cerebral vascular diseases and an efficient tool to study the therapeutic effect of medicine after surgery.

Objective:Studies suggested that some growth factors and their receptors play important roles in the treatment and prognosis of lung cancer.In this study we investigated the expression of epidermal growth factor receptor(EGFR),vascular endothelial growth factor(VEGF),and Ki-67 simultaneously in the tissues of aged patients with non-small cell lung cancer(NSCLC). Methods:The expression levels of EGFR,VEGF,and Ki-67 in 64 cases of aged patients with NSCLC were detected by immunohistochemistry staining.Results:The positive expression of EGFR,VEGF and Ki-67 in the tissue of lung cancer was 59.4%,57.8% and 92.2%,respectively.It is different significantly while compared with control group of benign tissues.The expression levels of EGFR and VEGF are correlated significantly with the pathologic type,differential degree,TNM stage and lymphatic metastasis.The expression level of Ki-67 is related to differential degree and TNM stage of lung cancer.Conclusion:The method of immunohistochemistry to detect multiple biological markers simultaneously is simple and feasible.It can supply important information for the staging and prognoses of patients with NSCLC,and more than that,it can also provide the theoretic basis for the individualized molecular targeting therapy of NSCLC.

Objective To investigate the effects of Krüppel like factor 4 (KLF4)on matrix metalloproteinase 9 (MMP9)ex-pression in hepatocellular carcinoma(HCC).Methods A total of 50 primary hepatocellular carcinoma samples and their correspond-ing adjacent tissues specimens were collected.The expression of KLF4 and MMP9 were detected by IHC,Western blot and qRT-PCR.After KLF4 gene was transfected into hepatocellular carcinoma cell line (HepG2 cell line),the expressions of KLF4 and MMP9 were conformed by qRT-PCR and Western blot.Migration and invasion of HepG2 cell line transfected by KLF4 were detec-ted by wound-healing assay and invasion assay.Results Compared to corresponding adjacent tissues,The expression of KLF4 was significantly lower in HCCs(P <0.05),and MMP9 expression was remarkably higher in HCCs(P <0.05).KLF4 over-expression inhibited the expression of MMP9 on the protein and mRNA levels.Wound-healing assay and invasion assay confirmed that KLF4 regulated cell invasion and migration through regulating MMP9 expression.Conclusion KLF4 showed low expression in HCCs,and MMP9 was overexpressed.Up-regulation of KLF4 could decrease the expression of MMP9 in HepG2 cell line,which inhibited inva-sion and migration.

This study is designed to obtain recombinant human acetylcholinesterase (rhAChE) and apply it in screening acetylcholinesterase inhibitors. The rhAChE was overexpressed in HEK293 cells transfected by plasmid of pCMV-AChE with the cationic liposome and rhAChE was found to be secreted into cell culture medium. AChE activity was assayed according to modified Ellman method to obtain kinetic parameters. IC so50 values for donepezil compounds of rhAChE were calculated to determine their activities of inhibition. The results showed that Km value was 151.9 micromol.L-1 donepezil inhibited rhAChE in a mixed competitive-noncompetitive way (Ki= 16.03 nmol.L-1, Ki = 18.36 nmol.L-1) and that most new compounds tested exhibited high activities of inhibition on rhAChE. The study suggests that rhAChE is available to be applied in screening AChE inhibitors in vitro.
Acetylcholinesterase ; genetics ; metabolism ; Cholinesterase Inhibitors ; analysis ; pharmacology ; HEK293 Cells ; Humans ; Indans ; analysis ; pharmacology ; Inhibitory Concentration 50 ; Kinetics ; Piperidines ; analysis ; pharmacology ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Transfection

Objective To compare the prevalence and correlation factors of chronic kidney disease (CKD) in urban and rural areas in Minhang district of Shanghai through the social economic and clinical data of the elderly population.Methods Jiangchuan Street and Pujiang town were randomly selected to represent the urban and rural population in Minhang district of Shanghai,respectively.Based on the over-60-year old people health examination program,6151 objectives with complete clinical-epidemiological data and bio-chemical index were investigated.The prevalence of CKD in urban and rural areas was compared,and the correlation factors for the urban and rural CKD were evaluated by multiple logistic regression analysis.Results (1) The survey objectives with an average age of (69.57+7.04) years,including 4345 cases of the city residents and 1806 cases of rural residents,were enrolled.The age structures of urban and rural showed differences,population over 80 years old account for 13.1％ of the rural total,significantly higher than 7.4％ in the urban population (P ＜ 0.001).(2) The prevalence rates of diabetes,hyperuricemia,hyperlipidemia and hyperlipidemia in urban residents were higher than those in rural residents,which were 26.4％ vs 13.7％,9.9％ vs 2.3％,53.7％ vs 37.4％,51.4％ vs 15.6％ (all P＜ 0.01).The awareness rates of kidney disease and hyperlipidemia showed significant differences in urban and rural areas,which were 32.9％ vs 44.2％,84.6％ vs 62.8％ (all P ＜ 0.01).Compared with those in rural areas,the treatment rates of hypertension and high blood lipids in urban residents were increased (all P ＜ 0.01).(3) The prevalence of CKD was 23.4％.Female CKD prevalence was higher than male,respectively 26.3％ and 18.5％ (P ＜ 0.01).In urban CKD prevalence was 22.2％,lower than 25.2％ in rural.The prevalence rate of hematuria in urban areas was lower than in rural areas,but the prevalence rate of decline in renal function was higher (all P ＜ 0.05).With the increase of age,the prevalence rate of CKD was increased (P ＜ 0.01).(4) Age (OR=1.072),smoking history (OR=1.543),previous history of kidney disease (OR=1.351),diabetes (OR=1.373),hyperuricemia (OR=2.498),obesity (OR=1.364),history of interventional therapy (OR=1.896) had positive correlation with CKD in city elderly population,while the higher education (OR=0.676,OR=0.604) and drinking (OR=0.585) had negative correlation (all P ＜ 0.05).Age (OR=1.032),female (OR=1.860) had positive correlation with CKD in rural elderly population (all P ＜ 0.05).Conclusions CKD has been a common chronic progressive disease of the aged in Minhang district.The prevalence of CKD is higher in urban areas than in rural.Age is a common factor for CKD in urban and rural.Previous smoking,history of kidney disease,diabetes,hyperuricemia,obesity,history of interventional therapy,education and drinking have correlation with urban CKD patients.Female has correlation with rural CKD population.

Objective To understand the prevalence,drug resistance and clinical features of children with acute lower respiratory tract infection(ALRI) caused by escherichia coli.Methods From Oct. 2005 to Oct. 2006,659 patients with ALRI who were admitted to hospital were chosen and their nasopharyngeal secretions were obtained and cultured.K-B disc diffusion for antibiotic susceptibility were performed for these clinical isolates.Results Among 659 patients,118 cases were isolated escherichia coli,the rate was 17.99% which had 90 boys and 28 girls.Eighty-seven of 118 E.coli strains were with extended-spectrum ?-lactamase(ESBLs),the rate was 73.73%.All of strains were sensitive to imipenem.For ESBLs-producing strains,the ratio of resistance tocefotaxime,ceftriaxone,cefuroxime,ampicillin,piperacillin were 78.81%,73.73%,73.73%,76.27%,78.81%,respectively.Conclusions The positive rate of ESBLs producing E coli in Kunming area is high and drug resistance is severe gradually.Imipenem can be the first selection for treatment on these infections.

BackgroundThe three-dimensional configuration of the nasolacrimal canal is highly variable with age,gender,and race.But enlargement of the nasolacrimal canal has sparsely been reported in the literature.Objective Computed tomography dacryocystography was performed in patients with unilateral congenital nasolacrimal duct obstruction and normal children to analyze the difference of bilateral nasolacrimal canal.MethodsThis is a retrospective study.Axial scanwith sagittalandcoronalreconstructionwas appliedin computedtomography dacryocystography.Diameters of bilateral nasolacrimal canal of 20 unilateral congenital nasolacrimal duct obstruction patients and 20 normal children were measured.Written informed consent was obtained from each child ' s parents before examination.ResultsThe lacrimal sac,nasolacrimal duct and the peripheral tissue were clearly exhibited by computed tomography dacryocystography.The diameters of the origination,the middle part and the distal end of affected nasolacrimal duct were(5.5±1.4),(5.3±1.2),(5.3±1.6) mm,and normal ones were(3.9±0.8 ),(3.5± 0.8 ),( 3.9± 1.3 ) mm,respectively.These results were statistically significant ( t =5.200,6.967,2.932,P＜ 0.05 ).There was no statistically significant difference in bilateral nasolacrimal canal of normal children (t =0.346,0.281,0.312,P＞0.05 ).Conclusions Computed tomography dacryocystography can image lacrimal passage and their peripheral tissues clearly.The affected nasolacrimal canal diameters of unilateral congenital nasolacrimal duct obstruction were much larger than the fellow sides.The pathogenesis of this phenomenon need much research.

Objective To explore the prevalence and independent influencing factors of arthritis in Chinese patients with systemic sclerosis(SSc).Methods A total of 248 SSc patients were enrolled in this prospective cross-sectional study in Peking Union Medical College Hospital.Demographic,clinical and laboratory data were compared in arthritis group and non-arthritis group.Results Of 248 Chinese SSc patients,mean age of onset was (37.1 ± 13.7) years old and disease duration was (7.4 ± 5.2) years.Overall prevalence of arthritis was 41.1％ (102/248).The presence of arthralgia(95.1％ vs 15.1％),muscle weakness(38.2％ vs 24.7％),short of breath (60.8％ vs 45.2％),and intestinal lung disease (64.7％ vs 49.3％) occurred more frequently in patients with arthritis than those without arthritis (P ＜0.05).The elevation of C reactive protein,anti-Sm and anti-cyclic citrullinated peptide were more common in arthritis group than in non-arthritis group(61.4％ vs 43.9％,11.4％ vs 2.5％ and 28.1％ vs 2/16,respectively).There were no obvious differences in the Rodnan score,digital ulcers,pulmonary arterial hypertension,and six-minute walk test between two groups(P ＞ 0.05).Conclusion This study reveals the significant prevalence of arthritis in a large cohort of SSc patients in China.Compared to non-arthritis patients,the SSc patients with arthritis have a more severe inflammation reaction and a higher rate of intestinal lung disease.

AIM: To investigate clinical effect of tongxinluo capsule in treating stable angina pectoris patients, and its influence on endothelial function, superoxide dismutase (SOD) and malondialdehyde (MDA). METHODS: One hundred and twenty-four stable angina pectoris patients were divided into three groups, isosorbide treatment group (41 cases), tongxinluo capsule treatment group (40 cases), tongxinluo and isosorbide treatment group (combined treatment group, 43 cases). The serum concentrations of nitric oxide (NO), endothelin-1 (ET-1), SOD and MDA were determined before and after treatment. The data in traetment groups were compared with that in normal control. RESULTS: The symptoms of 3 groups were significantly improved, and the total effective rate of tongxinluo capsule group and combined treatment groups were better than that in isororbide treatment group (85.00% and 88.37% vs 73.17%, P

Objective To study the correlation between microvessel density (MVD),isocitrate dehydrogenase 1 (IDH1) mutation and the malignancy of glioma,and its clinic significance.Methods The data and specimens of 40 patients with gliomas confirmed by surgery and pathology were collected.The relation between IDH1 mutation (detected by genetic sequence),MVD (detected by immunohistochemical coloration) and the malignancy of glioma was explored.5 cases of normal human brain tissues were used for comparative study.Results In normal brain tissue,Ⅰ,Ⅱ,Ⅲ,Ⅳ glioma,MVD counts were 8.12±1.64,25.10±1.27,27.00±1.98,42.80±10.75 and 56.50±5.23,respectively,and the overall difference was statistically significant (H =35.42,P ＜ 0.05).The MVD counts in low-grade glioma (Ⅰ-Ⅱ) and high-grade glioma (Ⅲ-Ⅳ) were 23.94±8.03 and 45.54±8.19,respectively,and the difference was statistically significant (t =-8.369,P ＜ 0.001).No mutation was found in normal human brain tissue,while in 20 cases of glioma specimens,there was IDH1 mutation with R132 as the mutation site and a MVD count of 31.11±13.47,and the other 20 cases of glioma specimens experienced no IDH1 mutation and the corresponding MVD count was 40.54±12.11.The difference of MVD counts of low-grade glioma and high-grade glioma was statistically significant (t =2.328,P=0.025).Conclusion MVD can be used as one of the histopathological grading metrics for glioma.IDH1 mutation occurs more frequently in grade Ⅱ and Ⅲ gliomas with R132 as the mutation site.