Objective To investigate the protective effect of ω-3 polyunsaturated fatty acid (ω-3 PUFAs ) on hypoxic ischemic brain damage(HIBD) in neonatal rats.Methods Eighteen pregnant rats were randomly divided into 3 groups, control group (6 pregnant rats, 26 new-born rats), sham operation group (6 pregnant rats, 24 neonatal rats), intervention group ( 6 pregnant rats , 28 newborn rats ) . Intervention group was administration with ω-3PUFAs (1 mg? g-1? d -1 ) by ga-vage , from 2 d after pregnancy to childbirth after 14 d.The rats in con-trol group and sham operation group were given the same amount of 0.9%NaCl.The newborn mice of 7 d old were made into HIBD model . Brain edema was measured using the gravimetric method .The expression of inducible nitric oxide synthase ( iNOS) and interleukin -6 ( iL-6 ) in brain tissue were detected by Western blot .The learning and memory function was assessed by the modified Morris water maze .Results Com-pared with control group , the newborn mice brain edema degree of inter-vention group was more ease , the difference was statistically significant (P<0.05).The expression of iNOS and interleukin -6 in newborn rat cerebral cortex of intervention group was more higher , the difference was statistically significant ( P <0.05 ) .The rats of learning and memory abilityininterventiongroupwasstronger,thedifferencewasstatisticallysignificant(P<0.05).Conclusion ω-3 polyunsaturated fatty acids may inhibit the expression of interleukin -6 and iNOS, and play a protective role in HIBD of newborn rat.

Objective To investigate the effect of erythropoietin ( EPO) on factor -related apoptosis and its ligand ( Fas/FasL ) expression in brain tissues of neonatal rats with hypoxic ischemic brain damage ( HIBD ) .Methods One hundred and twenty newborn rats ( 7 days years old ) were selected and randomly divided into 3 groups: sham group, hypoxic ischemic brain damage group ( model group ) and EPO treatment group ( test group ) , each group was further divided into five time points:6, 12, 24, 48 and 72 h after HIBD model was established. Eight rats in each group were killed at different time points.After which samples of brain tissues were harvested.Change of brain cell morphology was observed by HE staining and expression of Fas/FasL was observed by immunohistochemical staining.Results In model group, expressions of Fas and FasL reached the peak at 48 h after HIBD model was established and decreased at 72 h.Compared with sham group, the expressions of Fas and FasL in model group at each time point were much higher, and the difference was statistically significant ( P <0.05 ) .In test group, expressions of Fas and FasL at each time point were significantly lower than in model group ( P <0.05 ) .Conclusion Cell apoptosis in the brain tissue after hypoxic -ischemic damage could be remarkably inhi-bited by EPO through reduction of Fas and FasL expression in brain tissue of neonatal rats with HIBD.

Objective :To study effects of early rehabilitation after percutaneous coronary intervention (PCI) on car‐diac rehabilitation in patients with coronary heart disease (CHD).Methods :A total of 100 CHD patients undergoing PCI in our hospital were randomly and equally divided into routine nursing group and early rehabilitation group . Clinical indexes and incidence rate of major adverse cardiovascular events (MACE) after PCI ,scores of Morningside rehabilitative state scale (MRSS) ,self‐rating anxiety scale (SAS) ,self‐rating depression scale (SDS) ,social disabili‐ty screening schedule (SDSS) ,short‐form 36 health survey questionnaire (SF‐36) before and after treatment and pa‐tient satisfaction were compared between two groups before discharge .Results : Compared with routine nursing group ,there were significant reductions in first‐time get out of bed time 、 exercise time ,complete self‐care time , hospitalization time ,postoperative incidence rate of MACE (52% vs .18%) in early rehabilitation group , P＝0. 001 all ;After treatment ,compared with routine nursing group , there were significant reductions in scores of MRSS [ (5.87 ± 1.21) scores vs .(2.69 ± 1.01) scores] ,SAS [ (49.96 ± 3. 98) scores vs .(44.56 ± 3.12 ) scores] ,SDS [ (49.89 ± 3.85) scores vs .(45.38 ± 3.15) score] and each dimension score of SDSS ,and significant rise in SF‐36 score [ (89.76 ± 7.23) scores vs .(93.98 ± 8. 09) scores] and total satisfaction rate of patients (70% vs.90%) in early rehabilitation group , P＜0. 05 or ＜0.01. Conclusion :Early rehabilitation after PCI can significantly improve cardiac rehabilitation level ,psychological state ,social function and quality of life ,shorten postoperative recovery time and comprehensively improve patient satisfaction in CHD patients .

Objective To explore the potential value of knowing the relationship between congenital auricular deformities and middle ear malformations. Methods A total of 86 patients with congenital auricular deformities and middle ear malformations, including 51 males and 35 females, were admitted from January 2008 to December 2009 to the Eye Ear Nose and Throat Hospital of Fudan University. Fifty-eight with congenital auricular deformties were included. High-resolution CT (HRCT) data was obtained from each patient. The auricular deformities were classified into three grades using the Marx H classification system. The modified Jahrsdoerfer grading system was used to score the malformations using HRCT data.The correlation between the grades of auricular deformities and scores of middle ear malformations was analyzed using Spearman rank correlation analysis. Results The Marx H grades of congenital auricular deformties were 12 patients with grade Ⅰ , 25 patients with grade Ⅱ and 77 patients with grade Ⅲ, while their corresponding Jahrsdoerfer scores were 7.8 ± 2.4, 6.8 ± 2. 6 and 6.0 ± 2. 8, respectively. The statistical analysis suggested a trend of negative correlation between the Marx H grades of auricular deformities and the Jahrsdoerfer scores of middle ear malformations ( r = - 0. 2386, P = 0. 0106 ).Conclusion There was a trend to a negetive correlation between congenital auricular deformities and middle ear malformations.

OBJECTIVETo investigate the role of miR-221/222 in inhibiting endoplasmic reticulum stress-induced human hepatocarcinoma cells apoptosis.

METHODmiR-221/222 mimics and inhibitors were used to mimic or block the function of endogenous miR-221/222 respectively. Western blot and flow cytometry were used to test the effects of miR-221/222 on cell cycle and apoptosis under endoplasmic reticulum stress in human hepatocellular carcinoma cells.

RESULTSEndoplasmic reticulum stress resulted in miR-221/222 down-regulation in human hepatocellular carcinoma cells. miR-221/222 mimics and inhibitors inhibited and promoted respectively endoplasmic reticulum stress-mediated p27Kip1 induction. Moreover, p27Kip1 suppression not only resulted in reduction in the fraction of G1 phase cells, but also promoted the endoplasmic reticulum stress-mediated apoptosis in human hepatocellular carcinoma cells.

CONCLUSIONmiR-221/222 were downregulated by endoplasmic reticulum stress in human hepatocellular carcinoma cells, which subsequently protected human hepatocellular carcinoma cells against endoplasmic reticulum stress-induced apoptosis through p27Kip1 regulation.

Apoptosis ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Endoplasmic Reticulum ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; MicroRNAs ; metabolism

OBJECTIVETo prepare monoclonal antibody (McAb) against anti-CD3 ScFv for purifying and detecting serum anti-CD3 antibody concentration.

METHODSMcAb against anti-CD3 ScFv was prepared by hybridoma technique and used to prepare affinity chromatography column, which was used to purify anti-CD3 ScFv and Diabody [CD3 x Pgp] without E-tag. The binding activities of anti-CD3 ScFv, Diabody [CD3 x Pgp] without E-tag, and Diabody [CD3 x Pgp] purified by anti-CD3 affinity chromatography column or anti-E-tag affinity chromatography column against K562/A02 cell and Jurket cells were detected by fluorescence activated cell sorting (FACS) method. ELISA was used to identify the specificity of the McAb.

RESULTSMcAb against anti-CD3 ScFv specifically detected serum anti-CD3 ScFv without reacting with sera. The anti-CD3 ScFv purified by anti-CD3 affinity chromatography column and purified by anti-E-tag affinity chromatography column had the same specific binding activity with Jurkat cells. The positive binding rates of Diabody [CD3 x Pgp] without E-tag to K562/A02 and Jurkat cells were 89.87% and 83.95%, respectively. In the competitive binding experiments with K562/A02 and Jurkat cells, the binding rates of Diabody [CD3 x Pgp] without E-tag decreased to 56.30% and 43.78%, respectively.

CONCLUSIONThe McAb against anti-CD3 ScFv prepared in our lab can be used to purify and detect serum anti-CD3 antibody concentration.

Antibodies, Monoclonal ; biosynthesis ; immunology ; isolation & purification ; CD3 Complex ; immunology ; Cell Line ; Chromatography, Affinity ; Humans ; Hybridomas ; metabolism ; Jurkat Cells ; K562 Cells

OBJECTIVETo investigate whether the phosphorylation (functionally inhibitive) of eukaryotic initiation factor 2-alpha (eIF2-a) affects the molecular mechanism of cisplatin-induced cellular apoptosis in human hepatocellular carcinoma (HCC).

METHODSThe human HCC cultured cell lines SMMC-7221 and HepG2 were treated with cisplatin alone (controls; 24 h) or in combination with pre-transfection of a dominant-negative eIF2-a mutant (eIF2aS51A) or pre-exposure to an eIF2-a-specific phosphatase inhibitor (salubrinal) to decrease or increase the phosphorylation level, respectively. Changes in expression of apoptosis markers were quantitatively and qualitatively assessed by flow cytometry and western blot analysis. The significance of differences among groups was assessed by analysis of variance testing and of differences between groups was assessed by t-test.

RESULTSCisplatin treatment induced the appropriate functional-inhibitive phosphorylation of eIF2-a on serine 51. Cisplatin treatment (10 mg/ml) induced significant apoptosis in the eIF2aS51A pre-transfected SMMC-7721 (control: 21.7 +/- 1.5% vs. 50.7 +/- 2.1%, t = 19.454, P less than 0.05) and HepG2 (21.0 +/- 1.0% vs. 57.3 +/- 2.1%, t = 27.250, P less than 0.05). Salubrinal pre-treatment significantly inhibited the cisplatin (15 mg/ml)-induced apoptosis in SMMC-7721 (control: 50.3 +/- 2.5% vs. 16.3 +/- 2.1%, t = 18.031, P less than 0.05) and HepG2 (42.0 +/- 2.6% vs. 12.0 +/- 2.0%, t = 15.667, P less than 0.05).

CONCLUSIONPhosphorylation of eIF2-a may act to inhibit cisplatin-induced apoptosis of HCC.

Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Eukaryotic Initiation Factor-2 ; metabolism ; Humans ; Liver Neoplasms ; Phosphorylation

OBJECTIVETo explore the effects of survivin antisense RNA on taxol-induced apoptosis in leukemia cell line HL-60.

METHODSA survivin antisense eukaryotic vector pcDNA3-SVVas was transferred into HL-60 cells by electroporation. The live fraction was determined by trypan blue dye exclusion assay. Cell counting and MTT assay were performed to evaluate the sensibility of the transfected cells to taxol. Apoptosis was detected by DNA gel electrophoresis and nuclear staining.

RESULTSTwo positive cell clones, HL-60 SVVas and HL-60 neo were obtained. Compared to HL-60 and HL-60 neo cells, HL-60 SVVas cells growth was significantly reduced (P < 0.05). By MTT assay, the IC(50) of taxol to HL-60 SVVas, HL-60 neo and HL-60 cells were (14.4 +/- 1.87) ng/ml, (31.9 +/- 6.38) ng/ml and (32.0 +/- 3.52) ng/ml, respectively, the difference was significant by statistic analysis (P < 0.01). Agarose gel electrophoresis of genomic DNA from HL-60 SVVas showed typical DNA ladder, but DNA from HL-60 neo and HL-60 did not. Nuclei become condense in HL-60 SVVas cells.

CONCLUSIONSurvivin antisense RNA could enhance taxol-induced apoptosis in leukemia cell line HL-60. This may lay an experimental foundation for further research of gene therapy in leukemia.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Drug Synergism ; HL-60 Cells ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; genetics ; Neoplasm Proteins ; genetics ; Paclitaxel ; pharmacology ; RNA, Antisense ; genetics ; pharmacology ; Transfection

Objective To investigate the feasibility and efficacy on the outcome of patients with heart failure of integrated disease management program with heart failure clinic, patient education and telephone follow-up. Methods A total of 145 hospitalized patients with chronic heart failure and LVEF≤ 45% or patients with LVEF >45% and NT-proBNP > 1500 ng/L were divided into conventional group (re = 71) and interventional group (n =74). Patients were followed for 10 to 12 months. Results Baseline clinical characteristics, LVEF and dose of evidence-based medicine were similar between the 2 groups. During follow-up, the NYHA functional class was higher in conventional group than interventional group (3.2±0.5 vs 1.4±0.5, P <0.05) , and the LVEF deteriorated in the conventional group and improved from 34% to 40% in the interventional group. The proportions of self-monitoring of weight, blood pressure and pulse rate in the interventional group were significantly higher than those of conventional group (P < 0. 05). Among patients with systolic heart failure, 40% patients in the interventional group and 11 % patients in the conventional group achieved the target doses of β-blockers (P<0.05). Cardiovascular event rate of conventional group and interventional group is 91. 5% and 27. 0% respectively ( P < 0. 05 ). Conclusion Integrated disease management program with heart failure clinic, patient education and telephone follow-up can improve patient compliance to heart failure treatment, improve cardiac function and reduce cardiovascular event rate.

OBJECTIVEDuring 2003-2005, an outbreak of meningitis due to Neisseria meningitidis serogroup C occurred in China. With the aim to find strain clues result in the final epidemics, the ancestral strain 053442, a clinical isolate, and a carrier strain 053426 with different gene type were analyzed.

METHODSClinical strain 053442 and carrier strain 053426 were cultured on GC agar plates under the same condition. Two-dimensional electrophoresis was performed using the pH 3-10 nonlinear IPG strips of 24 cm length, and all the protein spots were identified by matrix-assisted laser desorption/ionization time of flight spectrometry.

RESULTS502 and 380 protein spots were identified in 053426 and 053442 respectively, relating to 266 and 202 different genes covering a wide range of cellular functions. The express volume and number of proteins involved in energy metabolism, protein synthesis and amino acid biosynthesis in 053426 were higher than in 053442. Virulence factor Opa, Opc and a series of proteins involved in pilus assembly and retraction were identified in 053442, which appear to be of primary importance in colonization and invasion of human cells. Compared to 053442, virulence protein species were less in 053426, with lower express volumes too. No Opa and Opc were detected in 053426.

CONCLUSIONSThe different protein expression profiles of the clinical strain 053442 and carrier strain 053426 in the present study provide some clues of the different pathogenicity of the two strains, which may account for result in the final epidemics.

Bacterial Proteins ; analysis ; Bacterial Typing Techniques ; China ; epidemiology ; Disease Outbreaks ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Meningitis, Meningococcal ; cerebrospinal fluid ; epidemiology ; microbiology ; Neisseria meningitidis, Serogroup C ; classification ; isolation & purification ; Proteome ; analysis ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization