Aim To study the effects of sodium ferulate on A?induced cognitive deficits and expressions of IL-1? and phospho-p38MAPK proteins.Methods Alzheimers disease model of rats was produced by intracerebroventricular injection of A?_(25-35)(10 ?g,once).Morris water maze was used to measure spatial memory performance.Nissl staining and immunohistochemical technique for glial fibrillary acidic protein(GFAP) were employed to determine the morphology of pyramidal neurons and astrocyte infiltration in hippocmpal CA1 regions.The levels of phospho-p38MAPK and IL-1? were determined by Western blot and ELISA method.Reverse transcription-PCR analysis showed changes in FasL mRNA.Results Intracerebroventricular injection of A?_(25-35)in rats resulted in spatial memory impairments shown by longer escape latency and decreased percentage of time spent in the target quadrant.These behavioral dysfunctions were accompanied by astrocyte activation and infiltration,increased IL-1? production and elevated FasL mRNA level,the loss of pyramidal neurons in hippocampal CA1,and the increase of phosphorylated p38MAPK.Oral administration of sodium ferulate(50,100,250 mg?kg~(-1)daily) and ibuprofen 15 mg?kg~(-1)daily markedly improved the memory impairment,attenuated pyramidal neuronal damage,and reversed the A?-induced increases in IL-1? and p38MAPK activation.Conclusion sodium ferulate prevents A?-induced neurotoxicity through suppressions of inflammatory response and the activation of p38MAPK.

OBJECTIVE To study the drug-resistant diversity of Gram-negative bacilli isolated from inpatients during recent five years.METHODS A total of 1 464 Gram-negative bacilli isolated were detected and retrospectively analyzed from 1999 to 2003.Antimicrobial susceptibility testing was performed using Kirby-Bauer method.RESULTS The resistance of Pseudomonas aeruginosa to piperacillin rised from 17.6% of 1999 to 79.2% of 2003,and that to ciprofloxacin rised from 4.3% of 1999 to 36.0% of 2003.The resistance of Escherichia coli to quinolones was above 50%,while to third-generation cephalosporins was 30-40%;the resistance of E.coli to piperacillin rised from 42.9% of 1999 to 68.9% of 2003,and that to ciprofloxacin rised from 40.0% of 1999 to 73.5% of 2003.The resistance of Acinetobacter to piperacillin rised from 31.2% of 1999 to 67.5% of 2003,and that to ceftriaxone rised from 36.0% of 1999 to 74.1% of 2003.The resistance of Serratia to ceftazidime,ceftriaxone,gentamicin,amikacin and piperacillin rised sharply.Imipenem was the most active antibiotic tested against Gram-negative bacilli.Cefoperazone/sulbactam and piperacillin/tazobactam also showed excellent activity against Gram-negative bacilli.CONCLUSIONS During recent five years,the resistance of the most common Gram-negative bacilli has increased rapidly.How to delay the resistance development of common strains become a global problem.

Objective To investigate the application and clinical significance of 24-hour multichannel intraaluminal impedance-pH (MII-pH) monitoring in gastroesophageal reflux disease (GERD).Methods Fiftythree patients with GERD were enrolled in this study according to the Montreal consensus (consulting for twice reflux a week or above) from July 2011 to June 2012.Patients were divided into erosive esophagitis (EE,n =25)group and non-erosive reflux disease (NERD,n =28) group after endoscopy and MII-pH monitoring.Fifteen healthy volunteers were recruited as the normal controls.The change of pH and MII-pH parameters were compared among the three groups and the significance of the change was investigated.Results Twenty-four-hour pH monitoring showed that all the acid reflux events in the EE and the NERD groups were significantly higher than those in the control group (P ＜ 0.05).MII-pH monitoring showed that the frequencies of total reflux,acid reflux,weakly acidic reflux and percentage of acid reflux in the GERD group were higher than in the control group (total reflux:83 (54,118) vs.62 (44,111) vs.42 (20,70),P =0.003 ; acid reflux:45 (25,79) vs.22 (11,45) vs.3(1,10),P =0.000 ;weakly acidic reflux:36(18,47) vs.43(21,82) vs.23(11,43),P =0.001 ;percentage of acid reflux:53％ (37％,81％) vs.32％ (13％,48％) vs.11％ (1％,23％),P =0.002).The frequency and percentage of acid reflux in the EE group were higher than those in the NERD group (P =0.000)The percentage of weakly acidic reflux in the NERD group was higher than in the EE group (66％ (43％,79％) vs.46％ (21％,57％),P ＜0.01).The frequencies of liquid reflux and mixed reflux in the GERD groups were higher than those in the control group (22 (12,40) vs.18 (12,26) vs.9 (4,18) ; 54 (39,79) vs.42 (25,77) vs.29(14,48) ;P ＜0.01).The frequency of gas reflux in the control group was higher than in the GERD group (86(56,207) vs.31 (14,62) vs.34 (15,119),P ＜ 0.01).The frequency and percentage of proximal reflux in the GERD group were significantly higher than in the control group (28(18,41) vs.16(12,34) vs.3 (2,9) ; 33％ (22％,49％) vs.29％ (22％,35％) vs.11％ (6％,22％),P ＜ 0.001).The percentage of symptom positive index in patients with acid reflux,non-acidic reflux and total reflux in the EE group were 36.0％ (9/25),20.0％ (5/25) and 56.0％ (14/25) respectively which were higher than in the NERD group (21.4％ (6/28),14.3％ (4/28) and 35.7％ (10/28)).Conclusion MII-pH monitoring can detect more reflux events.Acid reflux plays an important role in GERD.The detective rate of GERD will be elevated when combined with MII-pH monitoring in the diagnosis.MII-pH monitoring has a distinct advantage in diagnosing GERD.

BACKGROUND: Recently,the technique of isolation procedure of viable islets cell from large animals has been mature and used in clinical islet cell transplantation. Some study results indicate that low yields of pig islet cell isolations is probably not due to the presence of higher or more aggressive enzyme activities during the porcine isolation procedure. Therefore, the causative factors resulting in the inconsistent results should be sought for the intrinsic properties of the pancreatic islet and pancreas. The purpose of the study is to get more pancreatic islet cells for diabetic patients.OBJECTIVE: To investigate the effect of single enzymatic solution (SES)and multi-enzymatic solution(MES) on adult porcine pancreatic islet cell isolation,and to provide the theory base for diabetic patients rehabilitation.DESIGN: A completely randomized and controlled experiment based on animals.SETTING: General surgery and gerontology department in a university hospital.MATERIALS: The experiment was conducted in the Second Affiliated Hospital of Harbin Medical University from June 2003 to May 2004. Twenty consecutive random-bred adult pigs, 12 - 24 months old, weighing average 100 kg,were selected for experiment from Hada Slaughterhouse of Harbin.INTERVENTIONS: After slaughtering,the spleentic lobe of the pancreas was excised using sterile surgical gloves and surgical instruments. Each was immediately transported in 500 mL of sterile RPMI1640 solution at 4 ℃ to the laboratory for processing. After removal of fat peritoneum and superficial blood vessel,each pancreas was first immersed in 1:5 000 Liquor chlorhexidine for 3 minutes,then washed three times with cold RPMI1640, next mechanically minced into 1 mm×1 mm×1 mm fragments,weighed,and equally divided into two groups. Adult porcine islets were isolated with two different collagenase solutions(SES and MES).MAIN OUTCOME MEASURES: Islet count was performed with DTZ-staining. The viability was assessed by trypan-blue staining. Porcine islet insulin-secretory function was assessed by insulin content of cultured porcine islets and by insulin release. Islet morphological integrity was finally established by electron microscope examination.RESULTS: Overall islet cell content were statistically different between the two methods[(1 782 ±427) IE/g vs (1 293 ±451) IE/g,P<0.05]. No significant difference was found in viability,function or morphology of islet between MES and SES(P>0. 05).CONCLUSION: An average of isletisolated with MES method represents a uniquely massive yield in comparison with that by SES method. A good viability was also confirmed by static incubation and culture. The transplantation of those pancreatic islet cells can help to control the syndromes of diabetes.

Objective To investigate the bacterial spectrum and antimicrobial resistance of pathogens isolated from stool of acute diarrhea outpatients ,and provide scientific evidence for clinic rational use of antibiotics .Methods Bacteria was detected by conven‐tional feces culture method ,including separation and biochemistry appraisal sure strains .The predominant bacteria were conducted antimicrobial resistance testing in acute diarrhea outpatients .Results 544 stool specimens were collected from acute diarrhea outpa‐tients from January 2011 to December 2012 .The total positive rate was 17 .83% .Positive rates of Escherichia coli ,Salmonella , Campylobacter ,Vibrio parahaemolyticus ,Other Aeromonas ,Shiga Plesiomonas ,Shigella and Aeromonas hydrophila were 4 .78% ,3 .68% ,2 .57% and 2 .39% ,1 .84% ,1 .28% ,0 .92% and 0 .37% ,respectively .Salmonella ,Campylobacter and Vibrio parahaemolyticus were susceptible to Ofloxacin ,Amoxicillin ,Ceftazidime .They were different resistance to conventional antibiot‐ics ,which were commonly used by clinic ,and the most serious resistance are ampicillin and nalidixic acid .Conclusion Escherichia coli ,Salmonella ,Campylobacter and Vibrio parahaemolyticus are predominant bacteria pathogens .It is important to better under‐stand pathogens spectrum and antimicrobial resistance of bacteria for controlling infection in acute diarrhea outpatients .

Objective To evaluate the clinical impact of 18F-FDG PET/CT on patients with suspected cervical cancer recurrence. Methods Fifty-one cervical cancer patients, clinically suspected to have tumor recurrence during follow-up, underwent 18F-FDG PET/CT examination. 18 F-FDG PET/CT results were compared with those of conventional images, as referred to histopathology or clinical follow-up. Impacts of 18F-FDG PET/CT were evaluated based on documented changes of clinical management. Results In total, 43 patients were found to have positive lesions by 18F-FDG PET/CT, in which 40 were true recurrence,but 2 were pelvic abscess and 1 was radiation enterocolitis. Other 8 patients were found negative by 18F-FDG PET/CT and confirmed by pathology or follow-up. In patient-based analyses, the sensitivity, specificity, and accuracy of 18F-FDG PET/CT for the detection of tumor recurrence were 100% (40/40), 72. 73% (8/11),and 94.12% (48/51) respectively. In 7 patients, the clinical management was changed due to 18F-FDG PET/CT findings. Conclusion 18F-FDG PET/CT is an efficient tool for determining the recurrence of cervical cancer and instructing the clinical management.

AIM To investigate whether Aβ deposit in Alzheimer disease(AD) impairs signal transduction pathway responsible for neuronal survival.METHODSThe rats were randomly divided into six groups:control group and Aβ25-35 group,Aβ25-35+ibuprofen groups (7.5 and 15 mg·kg-1,respectively),Aβ25-35+ibuprofen+LY294002 group,and Aβ25-35+LY294002 group.Rats were given ibuprofen (7.5 and 15 mg·kg-1 daily,ig) for 3 weeks prior to and 1 week after icv single dose of Aβ25-35 (10 μL,1 mmol·L-1).LY294002 was injected icv 1 h before the injection of Aβ25-35.Seven days after Aβ25-35 injection,the hippocampal expressions of P53,Bax,Fas ligand (FasL),Bcl-2 proteins,phospho-Akt/PKB,and phosphorylated 70 ku ribosomal protein S6 kinase (p70S6K) and caspase 3 were determined in the brain tissue preparations from CA1 area with Western blot.The activity of caspase 3 was measured using a caspase 3 colorimetric activity assay kit.RT-PCR was used to show the change of p70s6k mRNA level.RESULTS Aβ25-35 icv injection significantly down-regulated phosphorylated Akt/PKB from 1.32±0.14 to 0.69±0.08 and p70S6K from 0.769±0.028 to 0.479±0.032 in hippocampal CA1 region.These changes were accompanied by increased expressions of the proapoptotic proteins P53,Bax,and FasL and decreased expression of the anti-apoptotic protein Bcl-2 in rat hippocampus.In addition,caspase 3 activity was significantly enhanced in hippocampal CA1 region in Aβ25-35-treated rats compared with control rats.Ibuprofen can reverse these Aβ25-35-induced changes.CONCLUSION Down-regulated anti-apoptotic PI3K/Akt/p70S6K signaling pathway induced by Aβ25-35 in rat hippocampus may contribute to the neuronal damage in AD.Ibuprofen prevents Aβ25-35-induced down-regulation of PI3K/Akt/p70S6K signaling pathway.

AIM To observe the neuroprotective effect and protective mechanisms of ibuprofen on amyloid β-protein fragment 1-40 (Aβ1-40)-induced neurotoxicity in rat hippocampus.METHODS Rats were given ibuprofen (15 mg·kg-1 daily,ig) for 3 weeks prior to icv single dose of Aβ1-40 (5 μL,1 mmol·L-1).Six hours after Aβ1-40 injection,Western blotting was used to determine the expressions of phospho-MAP kinase kinase (MKK)3/MKK6,phospho-p38 MAP kinase,phospho-MAP kinase activating protein kinase 2 (MAPKAPK2),heat-shock protein 27(Hsp27),procaspase 9,3,and 7 cleavage,and poly (ADP-ribose) polymerase (PARP) cleavage in hippocampal CA1 region.RESULTS Intracerebroventricular injection of Aβ1-40 induced an increase in phosphorylated MKK3/MKK6 and p38 MAP kinase expressions in hippocampal CA1.These increases,in combination with reduced phospho-MAPKAPK2 and phospho-Hsp27 expressions,mediated Aβ1-40-induced the activation of caspases cascades.Ibuprofen (15 mg·kg-1·d-1,3 weeks) significantly prevented Aβ1-40-induced increases in phosphorylated MKK3/MKK6 and p38 MAP kinase expressions.In addition,Aβ1-40-induced decreases in phosphorylated MAPKAPK2 and Hsp27 expressions were abrogated by ibuprofen.Aβ1-40-induced changes in activation of caspases cascades were inhibited by ibuprofen.CONCLUSIONIbuprofen prevents Aβ1-40-induced neurotoxicity through suppression of phosphorylated MKK3/MKK6 and p38 MAP kinase expressions and the up-regulation of phospho-Hsp27 expression.

AIM To explore the mechanism of amyloid β-protein fragment 25-35(Aβ25-35)-induced inflammation and apoptosis in rat hippocampus in vivo by studying mitogen-activated protein kinase (MAPK) signaling pathway and the protective effect of anti-inflammatory drug ibuprofen. METHODS Rats were given ibuprofen (7.5 mg·kg-1 daily, ig) for 3 weeks prior to and 1 week after icv single dose of Aβ25-35 (10 μL, 1 mmol·L-1). Seven days after injection, Nissl staining and immunocytochemical technique were employed to determine the morphology of pyramidal neurons and astrocyte infiltration in hippocampal CA1. The expressions of IL-1β, extracellular signal-regulated kinase (ERK), p38 MAPK, PKC, and caspase-3 were determined by Western blot. Reverse transcription-PCR analysis showed changes in IL-1β mRNA level. RESULTS Intracerebroventricular injection of Aβ25-35 elicited astrocyte activation and infiltration and caused a strong inflammatory reaction characterized by increased IL-1β production and elevated IL-1β mRNA level. The inflammatory reaction was accompanied by the loss of pyramidal neurons in hippocampal CA1. The phosphorylation of p38 MAPK was significantly increased, on the other hand, the phosphorylation of ERK was significantly reduced and these were coupled with the increase of caspase-3 expression in hippocampal CA1. Ibuprofen (7.5 mg·kg-1 daily, 4 weeks) significantly reduced Aβ-induced IL-1β expression, caspase-3 expression and p38 MAPK activation. The loss of pyramidal neurons was also significantly attenuated by treatment with ibuprofen. CONCLUSION The activation of p38 MAPK and the down-regulation of ERK play a pivotal role in the inflam-matory response and apoptosis evoked by Aβ25-35 in vivo, which can be prevented by ibuprofen.

Chronic myeloid leukemia (CML) is characterized by the accumulation of active BCR-ABL protein.Imatinib is the first-line treatment of CML;however,many patients are resistant to this drug.In this study,we aimed to compare the differences in expression patterns and functions of time-series genes in imatinib-resistant CML cells under different drug treatments.GSE24946 was downloaded from the GEO database,which included 17 samples of K562-r cells with (n=12) or without drug administration (n=5).Three drug treatment groups were considered for this study:arsenic trioxide (ATO),AMN107,and ATO+AMN107.Each group had one sample at each time point (3,12,24,and 48 h).Time-series genes with a ratio of standard deviation/average (coefficient of variation) ＞0.15 were screened,and their expression patterns were revealed based on Short Time-series Expression Miner (STEM).Then,the functional enrichment analysis of time-series genes in each group was performed using DAVID,and the genes enriched in the top ten functional categories were extracted to detect their expression patterns.Different time-series genes were identified in the three groups,and most of them were enriched in the ribosome and oxidative phosphorylation pathways.Time-series genes in the three treatment groups had different expression patterns and functions.Time-series genes in the ATO group (e.g.CCNA2 and DAB2)were significantly associated with cell adhesion,those in the AMN107 group were related to cellular carbohydrate metabolic process,while those in the ATO+AMN107 group (e.g.AP2M1) were significantly related to cell proliferation and antigen processing.In imatinib-resistant CML cells,ATO could influence genes related to cell adhesion,AMN107 might affect genes involved in cellular carbohydrate metabolism,and the combination therapy might regulate genes involved in cell proliferation.