Objective: To investigate the expression of nuclear factor-κB (NF-κB) subunits P50 and c-Rel protein in primary cortical neurons of Wistar rats at different time points of oxygen glucose deprivation/ reoxygenation(OGD/R). Methods: The neurons dissociated from the cortex of the neonatal rats were primary cultured and were identified by immunocytochemistry. OGD/R model was established. The study was divided into 6 groups according to different processing methods,including normal group,OGD 4 h treated,OGD 4 h/R 2 h treated,OGD 4 h/R 6 h treated,OGD 4 h/R 12 h treated and OGD 4 h/R 24 h treated groups. The expression of NF-κB P50 and c-Rel protein in neurons was examined by immunocytochemistry method and Western blotting. Results: (1) Immunocytochemistry detection targeting neuron specific enolase (NSE) and beta-III tubulin confirmed that the cultured cells were neurons. (2) The expression of NF-κB P50 protein was significantly higher in OGD 4 h group than in control group(P<0.05); the expression continued to increase in OGD 4 h/R 2 h andOGD 4 h/R 6 h groups, and reached its peak 6 h after reoxygenation (P<0.01),then began to decrease,but the expression in OGD 4 h/R 12 h group was significantly different from that of the control group (P<0.05); there was no significant difference between OGD 4 h/R 24 h group and control group. (3) The expression of NF-κB c-Rel protein was similar between OGD 4 h group and the control group; the expression increased in OGD 4 h/R 2 h and OGD 4 h/R 12 h groups and reached its peak 12 h after reoxygenation(P<0.01),and did not recover to the normal level in OGD 4 h/ R 24 h group (P<0.05). Conclusion: Oxygen glucose deprivation/reoxygenation can activate NF-κB subunits P50 and c-Rel in the primary cortical neurons in rats in a time-associated manner.

Aortic Coarctation ; complications ; Ductus Arteriosus, Patent ; complications ; Humans ; Infant ; Pulmonary Artery ; abnormalities

Objective:To develop the uniformity models for felodipine sustalned-release tablets from 3 manufacturers by NIRS in order to study the difference in the preparation technology and detect and screen the tablets quickly by the robust, accurate and repre-sentative models. Methods:The uniformity models for the tablets from 3 manufacturers among 6 manufacturers with evaluative casual inspection were established by NIRS. Region 4 000-9 000 cm-1 was chosen as the modeling section, and the first derivative plus vector normalization was used as the preprocessing method. Results:The uniformity models for the tablets from the three manufacturers was established and used to predict the samplings from the six manufacturers. The prediction success rate was 100%. Conclusion: NIRS can be used to identify felodipine sustalned-release tablets from different manufacturers quickly and study the preparation technology.

Objective To investigate the relationship between cysteine-rich protein 61 ( Cyr61 ) and phosphatidylinositol 3-kinase ( PI3K ) signal pathway on cell proliferation and apoptotic in human ovarian carcinoma cells.Methods Recombinant human Cyr61 (rhCyr61) was pretreated with ovarian carcinoma cells.The expression of Cyr61 protein was detected by confocal spectral microscopy.Then treated the ovarian carcinoma cells with PI3K transduction inhibitors (LY294002) for 24 hours.Cell apoptosis was detected by flow cytometry (FCM).Cell viability was determined by methyl thiazolyl tetrazolium (MTT) method.The mRNA expressions of Cyr61,the protein levels of protein kinase B ( PKB),phospho-PKB and Cyr61 were assaved by real time-PCR and western blot analysis,respectively.Results The Cyr61 and phospho-PKB protein expression in two ovarian carcinoma cells (OV2008 and OVCAR-3 ) were increased in rhCyr61pretreated group.The decreasing of cell apoptosis [ ( 1.4 ±0.9)％,(2.1 ± 1.0)％ ] and increasing of cell proliferation [ ( 124.0 ± 1.8)％,( 133.0 ±2.2)％ ] was detected in the same time,compared with negative control group,there were significant difference ( P ＜ 0.05 ).After exposed to LY294002 for 24 hours,the apoptosis rate of OV2008 and OVCAR-3 in pretreated with rhCyr61 group exposed to LY294002 were (21.1 ± 1.6)％ and (26.4 ± 1.5 )％,respectively.Cells viability [ (59.0 ± 2.3 )％,(51.0 ± 2.0)％ ]was also significantly decreased in OV2008 and OVCAR-3 pretreated with rhCyr61 cells.Meanwhile,the mRNA expressions of Cyr61 (3.2 ± 0.8,6.2 ± 1.1 ) and the protein levels of phospho-PKB and Cyr61 were greatly decreased.Compared with negative control group,there were significant difference in OV2008 and OVCAR-3 cells (all P ＜ 0.0l ).Conclusions The activation of PI3K intracellular signaling pathways may lead to up-regulation of Cyr61 expression.Block PI3K signal pathway could significantly inhibit the expression of Cyr61,and may promote the apoptotic effects and inhibit the cell growth of ovarian carcinoma cells.

OBJECTIVE:To systematically review the effectiveness and safety of methotrexate(MTX)in the treatment of rheuma-toid arthritis,and provide evidence-based reference for clinical treatment. METHODS:Retrieved from Medline,EMBase,Cochrane Library,PubMed,CJFD,CBM,VIP and Wanfang Database,randomized controlled trials(RCT)about MTX in the treatment of rheu-matoid arthritis were collected,Meta-analysis was performed after data extraction and quality evaluation. RESULTS & CONCLU-SIONS:33 effectiveness evaluation were included,involving 8 253 patients,and 53 safety evaluation were included,involving 4 803 patients. Results of analysis show,the efficacy of MTX was not higher than leflunomide in the treatment of rheumatoid arthritis,superi-or to cyclosporine A,and similar to sulfasalazine. In addition,MTX shows similar efficacy with etanercept(7.5-20 mg per week),goli-mumab,adalimumab or rituximab,but less effective than trastuzumab. The incidence of adverse reactions of MTX is high,but mainly mild or moderate,and the most common adverse reactions are gastrointestinal symptoms. Oral administration of MTX is more secure than intramuscular injection and subcutaneous injection.

Objective Chiral recognition for the glimepiride and glimepiride-cis-isomer by applying three amino acid (D-Lysine,L-Glutamine and L-Tyrosine) as chiral probes based on electrospray ionization mass spectrometry (ESI-MS) was achieved.Methods glimepiride/glimepiride-cis-isomer solutions were mixed with three amino acid solutions.The complex was extracted by ESI-MS and then the fragmentation abundance was investigated applying collision induced dissociation (CID) by MS/MS,which is the basis of chiral recognition for glimepiride and glimepiride-cis-isomer.Results Chiral recognition effect was achieved with the recognition rate (R) 1.61,2.92 and 2.17 for D-Lysine,L-Glutamine and L-Tyrosine respectively.Conclusion 3 kinds ofchiral amino acids were used as probes to distinguish between stereoisomers,and rapid identification of glimepiride and glimepiride cis isomer by mass spectrometry come true for the first time.

Objective To explore the effect of Curcumae on acute hepatic injuried mice.Methods The model mice of acute hepatic injury were established with intraperitoneal injecting 0.2% carbon tetrachloride (CCl4) 0.2 ml/10mg. 48 mice were randomized into 6 groups: model group, control group, Ganlixin group (25 mg/kg) and three treating groups of Curcumae which were administrated by gavage at the doses of 2.5 g/kg, 5 g/kg, 10 g/kg once everyday.7 d later, all mice were put to death. The liver index, the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) in serum, the activities of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in hepatic tissue were observed. Pathological change was observed under HE staining. Results Compared with the model group, Curcumae could decrease the liver index and the activities of ALT and AST in serum, MDA in hepatic tissue significantly (P<0.05), and increase the activities of SOD (P<0.05). The pathological changes were less in three treating groups than that of the models. Conclusion Curcumae can protect the liver from acute hepatic injuried induced with CCl4 in mice.

Objective To investigate the effect of great omentum combined with medical obturation glue on preventing thoracic cavity anastomotic leakage.Methods From August 2008 to September 2012,560 patients with esophageal gastric cardial carcinoma were enrolled and divided into two groups:the regular group (n =280) and the experimental group (n =280).In the regular group,anastomosis was reinforced with interrupted mattress sutures after esophageal gastric anastomosis was stapled.In the experimental group,anastomosis was covered with great omentum and medical obturation glue was sprayed to conglutinate after reinforced with interrupted mattress sutures.After that,gastric corpus was fixed upon the thoracic aorta and posterior chest wall.The clinical effects of the two groups were compared.Results Intrathoracic anastomotic leakage occurred in 8 cases (2.86％(8/280)) of the regular group,including 7 cases with symptomatic leakage and 1 case with asymptomatic loculate leakage.Seven patients were cured with conservative treatment and 1 patient with severe infection left hospital without cure.Average length of hospital stay was (55.6 ± 30.5) days postoperatively.Anastomotic stenosis occurred in 11 patients (3.93％,11/280).In the experimental group,one patient (0.36％,1/280) with asymptomatic loculate leakage was hospitalized for 20 days,and finally cured and discharged.8 cases with anastomotic stenosis occurred in the experimental group (2.86％,8/280).There was statistic difference in the rate of intrathoracic anastomotic leakage between the two groups (P =0.044),but there was no statistic difference in anastomotic stenosis between the two groups (P =0.484).Conclusion The technique of great omentum combined with medical obturation glue for preventing thoracic cavity anastomotic leakage,which is easy to perform,can obviously decrease the occurrence and attenuate the symptom of intrathoracic anastomotic leakage,and anastomotic stenosis increases unobviously.It also can shorten the length of hospital stay and is worthy of clinical promotion.

Objective To investigate the occurrence circumstance of acute stress disorder(ASD),clinical features and related risk factors,in the officers and soldiers who had gone to Beichuan county to provide disaster relief.Methods We used the self-general questionnaire,post-traumatic stress symptoms self-rating scale(PCL-C),perceived social support scale(PSSS),and coping style questionnaires(SCSQ)to assess the 126 subjects,combining with semi-structured interviews to determine and compare the ASD group and control group,and then analyzed the related factors.Results The prevalence rate of ASD officers and soldiers was 13.49%,with main symptoms of repeatedly breaking into painful memories,nightmares,reproducing the scenes of traumatic events,strong trouble of psychological pain,decreased interest,sleep disorders and emotional instability.These main symptoms of ASD interacts each other,and were reduced with positive response and good social support.Conclusion In the phase of acute stress(one month),the officers and soldiers taking part in earthquake relief have symptoms of ASD,so we should strengthen the necessary psychological assessment and comprehensive psychological intervention means,in order to slow down psychological trauma after disaster,prevent and reduce the possibility that ASD turns into post-traumatic stress disorder(PTSD).

Aim To study the effects of cycle arrest and molecular mechanism in human leukemia HL-60 cells induced by diallyl disulfide ( DADS ) . Methods Cell count, colony formation in soft agar experiments and flow cytometry analysis were employed to observe the DADS-induced cell growth inhibition and the effect of cycle arrest in HL-60 cells. The expressions of Chk1/2 and its downstream element in HL-60 cells were detected by Western blot. Results Cell count revealed that population doubling time increased to 35. 03 h and 71. 82 h, respectively, from 19. 14 h in HL-60 cells treated with 60 and 120 μmol·L-1 DADS ( P<0. 05 ) . Colony formation in soft agar experiments showed that colony formation inhibition rate of HL-60 cells exposed to 30, 60, 90 and 120μmol·L-1 DADS increased to 35. 06%, 62. 10%, 93. 79% and 99. 35%, respectively ( P<0. 05 ) . Flow cytometry a-nalysis exhibited that HL-60 cells treated with 60 and 120 μmol · L-1 DADS for 24 h and 48 h arrested in G2/M phase in a concentration-and time-dependent manner ( P <0. 05 ) . Western blot disclosed that the expression of p-Chk1 increased in a time-dependent manner ( P <0. 05 ); however, Chk1, Chk2 and p-Chk2 were not changed in HL-60 cells treated with 60μmol·L-1 DADS (P >0. 05). The expression of Cdc25C, CyclinB1 and CDK1 decreased after treated with 60 μmol·L-1 DADS in a time-dependent manner ( P<0. 05 ) , but the expression of 14-3-3 protein did not change ( P>0. 05 ) . Conclusion DADS can in-hibit the proliferation of HL-60 cells, and induce G2/M arrest through Chk1/Cdc25 C/CyclinB1/CDK1 path-way.