Objective To investigate the effect of 5-aza-2'deoxycytidine (5-Aza-CdR) on proliferation and expression of RASSF1 A gene in human ovarian cancer cell line SKOV3 and 3AO.Methods SKOV3 and 3AO cells were treated with different concentrations (0.5,5,50 μmol/L) of DNA methyltransferase inhibitor 5-Aza-CdR.RT-PCR and Western Blot were adopted to detect expression of mRNA and protein of RASSF1A gene before and after treatment with 5-Aza-CdR respectively.Results Compared with control group,when the 5-Aza-CdR concentration was 0.5,5,50 μmol/L after drug treatment,human ovarian cancer cells could significantly inhibit tumor cell growth; SKOV3 and 3AO cells in control group were observed weaker expression of RASSF1A mRNA.After treated with 5-Aza-CdR,the expressions of RASSF1A mRNA were observed increased with the increase of the drug concentration.After treated with different concentration of 5-Aza-CdR,the expressions of RASSF1A mRNA treated with 0.5 μmol/L 5-Aza-cdR was lower than those treated with 5 and 50 μmol/L 5-Aza-cdR (t =-8.866,P =0.01 ; t =-12.256,P =0.000).However,expressions of RASSF1A mRNA treated with 5 and 50 μmol/L 5-Aza-cdR respectively showed no statistical significance (t =0.431,P =0.689).Expressions of RASSF1A protein treated with 0.5 μmol/L 5-Aza-cdR and 5 μmol/L 5-Aza-cdR didn't show statistically significant (t =-1.586,P =0.188).Conclusion Expressions of RASSF1A mRNA and protein in SKOV3 and 3AO cells were evidently enhanced.As one kind of methyltransferases inhibitors,5-Aza-CdR can inhibit ovarian cancer cell line SKOV3,3AO growth through the RASSF1A promoter methylation,and thus promote their apoptosis.

Objective To evaluate the effectiviness and safety of rituximab in the treatment of immune thrombocytopenia .Meth-ods Standard doses and small doses of rituximab alone or combined therapy with other methods were used in 2 cases with immune thrombocytopenia who diagnosed and treated for the first time ,and 6 cases who diagnosed with refractory recurrent immune throm-bocytopenia .The recent curative effect ,adverse reactions were analyzed .Results The recent curative effect was well in the 2 cases who diagnosed and treated for the first time ,but 1 case recurrenced after 6 months .4 cases of effective in the 6 cases who diagnosed with refractory recurrent ,in which 1 case recurrenced after 2 years ,2 cases had no effect .All of the patients have no obvious adverse reaction in the near future .Conclusion Rituximab treatment of immune thrombocytopenic ,it is of good curative effect ,good safety and tolerability .

ObjectiveTo compare the effect of hospital initiative management or patients self management in asthma control and pulmonary function, and improve the control level of asthma. Methods Forty moderate asthma patients enrolled successfully from July to December in 2009 were divided into 2 groups by random digits table:hospital initiative management group (group A) and patient self management group (group B) with 20 cases each, and received the asthma treatment with hospital initiative management or patient self management for 1 year. The acute attack time, emergency visit time, hospitalization time,pulmonary function and Saint George respiratory questionnaire (SGRQ) were compared between two groups.ResultsAfter 1 year management, there were 19 patients in good compliance, satisfaction score was (9.300 ± 0.801 ) scores, 13 total control, 6 partial control and 1 uncontrol in group A, while there were 11 patients in good compliance, satisfaction score was (7.800 ± 1.542) scores, 6 total control, 8 partial control and 6 uncontrol in group B. The compliance, satisfaction and control in group A were much better than those in group B (P＜0.01 or ＜0.05). The forced expiratory volume in 1 second (FEV1)[(2.56 ±0.30) L]and peak expiratory flow (PEF)[(6.26±0.39) t/s]were elevated while SGRQ[(21.55 ±6.35) scores]in group A were better than those in group B[(2.38 + 0.31 ) L, (5.83 ± 0.52 ) L/s,(29.80 ± 12.04) scores](P ＜ 0.05 or ＜ 0.01 ). ConclusionThe compliance, asthma control, pulmonary function and respiratory quality are improved by hospital initiative management, so it is helpful to promote this management model in China via a close cooperation between general hospital and community hospital.

Objectives To evaluate the effect of N-Acetylcysteine on the airway inflammation and remodeling of obese asthma mice with high-fat diets. Methods Forty female C57BL/6J mice were randomly divided into 4 groups, asthma group (A), obese asthma group (B), treatment group (C) and normal control group (D). Group A were sensitized and challenged by ovalbum (OVA) and normal diets. Group B were sensitized and challenged as group A but fed with high-fat diets, while group C were sensitized, challenged and fed as group B, but administrated N-Acetylcysteine 200 mg/kg. d from the third week after challenge. The cells in BALF were counted and classified after staining, IL-6 and 8-iso-PGF2α(8-iso-PGF2α) in lung tissues were detected by ELISA. WAt, WAm, Pbm, as the remodeling indices, measured in lung pathological section. All parameters were compared among 4 groups. Results In comparison with group D, the leukocytes and EOS in BALF, WAt/Pbm, WAm/Pbm in lung section were increased as well as IL-6 and 8-iso-PGF2α in lung tissue elevated in group A and group B, while the maximum changes were observed in group B (P ＜0. 05). After NAC treatment, the IL-6, 8-iso-PGF2α and WAt/ Pbm were decreased significantly (P ＜0. 05). Pearson correlation analysis showed that WAt/Pbm and IL-6 were in positive correlation with 8-iso-PGF2α (r =0. 817, 0. 737, P ＜0. 01). Conclusions N-Acetylcysteine can alleviate the airway inflammation and remodel reaction of asthma by a substantial inhibition of the oxidative stress reaction in lung.

Objective To investigate the blood glucose changes in critically ill patients taken continuous renal replacement therapy(CRRT)before treatment and at different time points after treatment. Methods Sixtyseven critically ill patients were enrolled in the study,of whom 13 cases were treated conventionally and assigned as pre-treatment control group,while the other 54 cases undertook CRRT were assigned as CRRT group. The CRRT group were further sub-divided into four groups according to CRRT time((CRRT time ＜6 h group(n = 18),CRRT time of 6 - 12 h group(n = 18),CRRT time ＞ 12 h group(n = 18);or sub-divided into two groups according to circulating blood temperature(≤ 36. 5 ℃ group(n =27)and ＞ 36. 5 ℃ group(n =27));or sub-divided into two groups by sugar substitute fluid replacement solution as the formula group(the standard level of sugar content was 6. 9 mmol/L(n = 27)and sugar-free replacement fluid group(n = 27);or sub-divided into two groups according to the Glasgow Coma score(GCS)(GCS ≤ 8 points group(n = 24)and GCS ＞ 8 group(n = 30). Blood glucose level at different time points were measured. Results The longer CRRT cycle time,lower temperature,sugar substitute fluid replacement fluid worse GCS score corresponded to more dramatic blood glucose fluctuation at different time points in CRRT group,and it took longer time for them to recover to normal blood sugar level Conclusions During the CRRT process insulin secretion was suppressed and led to increase of blood sugar. In most cases the blood glucose returns to normal after CRRT treatment over time,but a small number of cases require clinical intervention.

Objective To study the anti-inflammatory effects of idazoxan (IDA) on endotoxin lipopolysaccharide (LPS) challenged mice in vivo and activated macrophages in vitro,and explore its potential molecular mechanisms.Methods To do the experiments in vivo,30 adult male C57BL/6 mice were randomly divided into control group,model group,and low,medium and high doses IDA groups (IDA-L,IDA-M,and IDA-H groups),n =6 in each group.The inflammatory model was reproduced by intraperitoneal injection of LPS 10 mg/kg,and the control group was injected with the same amount of normal saline.The IDA groups received LPS (10 mg/kg) and IDA 0.3,1.0 and 3.0 mg/kg,respectively.The blood samples of mice in each group were collected at 6 hours after the reproduction of the model.For the in vitro experiments,primary peritoneal macrophages were collected from 20 adult male C57BL/6 mouse cells and they were divided into control group,LPS group (10 mg/L) and LPS+IDA-L,IDA-M,IDA-H groups (10 mg/L LPS + 5,25,100 μmol/L IDA,respectively).Cell culture supernatants were collected at 24 hours after the reproduction of the model.Detection methods:enzyme linked immunosorbent assay (ELISA) was used to determine the levels of serum tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6),monocyte chemotactic protein-1 (MCP-1) and nitric oxide (NO).Western Blot was used to determine the effect of IDA on the expression levels of nuclear factor-κB (NF-κB) in macrophages.Results ① For the in vivo experiment,the serum levels of TNF-α and IL-6 were significantly elevated in the model group as compared with those in the control group [TNF-o (ng/L):403.96 ± 40.98 vs.17.50 ± 8.68;IL-6 (ng/L):61 400.31 ± 7 826.61 vs.2 436.30 ± 448.89;both P ＜ 0.01].IDA treatment could inhibit the elevation of inflammatory cytokines in a dose-dependent manner,with the most significant decrease in LPS+IDA-H group [TNF-α (ng/L):170.09 ± 28.53 vs.403.96 ± 40.98,IL-6 (ng/L):16 570.81 ± 1 083.65 vs.61 400.31± 7 826.61;both P ＜ 0.01].② For the in vitro experiment,the levels of TNF-α,IL-6,MCP-1,and NO secreted by LPS-stimulated macrophages were distinctly higher in the LPS group than those in the control group [TNF-α (ng/L):7 259.14 ± 320.70 vs.28.50±27.08,IL-6 (ng/L):14809.60±5852.73 vs.1 113.47±465.53,MCP-1 (ng/L):20847.37± 1 788.33 vs.447.37± 395.69,NO (μmol/L):1 900.00 ± 144.31 vs.603.03 ± 102.18;all P ＜ 0.01].However,IDA intervention could lower the secretion of TNF-α,IL-6,MCP-1 and NO in a dose-dependent manner,with the most notable decrease in the LPS+IDA-H group [TNF-α (ng/L):784.40±281.90 vs.7259.14±320.70,IL-6 (ng/L):1 802.96± 1 534.18 vs.14 809.60± 5 852.73,MCP-1 (ng/L):2005.26± 1 534.28 vs.20847.37 ± 1 788.33,NO (μ mol/L):654.54± 150.21 vs.1 900.00 ± 144.31;all P ＜ 0.05].In addition,IDA at the concentration of 100 μmol/L could promote the translocation of NF-κBp65 in macrophages into the nucleus 15 minutes early and lead to increased NF-κBp65 expression (gray value:18.70 ± 2.29 vs.1.09 ± 0.36,P ＜ 0.05),hut significantly reduce the expression levels of NF-κBp50 in the nucleus at 45 minutes after treatment (gray value:1.99 ± 0.14 vs.2.94 ± 0.54,P ＜ 0.05).Conclusions IDA could significantly reduce inflammation of mice challenged with LPS and inhibit inflammatory cytokines and mediators secreted by macrophage in a dose-dependent manner.High concentration of IDA (100 μmol/L) exhibited the greatest anti-inflammatory effects.The anti-inflammatory effect of IDA may be worked through NF-κB signaling pathway.

Objective To detect the effect of deep hypothermia on the function of mitochondria in hippocampus after global ischemia in rats and to explore the protection mechanism. Methods The animal model of cardiopulmonary bypass (CPB) was established in rats that were then randomly divided into three groups,ie,control group,normothermia ischemia group and hypothermia ischemia group,eight rats per group.The mitochondria was extracted from the hippocampus of each rats for observing the mitochondrial respiratory function,the activities of succinate dehydrogenase (SDH),the cytochrome oxidese(CCO),the lnembrane fluidity and the content of intramitochondria free calcium and MDA. Resuits The content of intramitochondria free calcium and MDA in the normothermia ischemia group was increased significantly compared to the control group and that in the hypothermia ischemia group wag decreased significantly compared with the normothermia ischemia group(P<0.05).Respiratory state Ⅲ (R3),respiratory state IV(R4),P/O ratio and oxidative phosphorylation (OPR) in the normothermia ischemia group were decreased significantly compared to the control group (P<0.05).R3,R4,P/O ratio and OPR in the hypothermia ischemia group were increased significantly compared with the normothermia ischemia group (P<0.05).Membrane fluidity in the normothermia ischemia group wag decreased significantly compared to the control group (P<0.01),while that in the hypothermia ischemia group was increased significantly compared with the normothermia ischemia group(P<0.05).The activities of SDH and CCO in the normothermia ischemia group were decreased significantly compared to the control group (P<0.01),while those in the hypothermia ischemia group were increased significantly compared with the normothermia ischemia group (P<0.05). Conclusion Profound hypothermia exerts a protective effect on the function of mitochondria in the hippocampus after global ischemia in rats.

High mobility group protein B1 (HMGB1) is the most representative substance in the alarmins family, it is actively or passively release to extracellular by the activation of monocyte/macrophage and the dead cells, and then it stimulates the production of a variety of inflammatory mediators, and increases the organism's inflammatory response through relevant receptors signaling pathways. In recent years, its concentration can reflect the severity of inflammation and injury and was related to the prognosis, HMGB1 has won more and more attention in the development of sepsis. By reviewing the study of HMGB1 in sepsis pathogenesis, signal pathway and reversal measures, it was found that HMGB1 was considered as an important inflammatory mediators and warning signal involved in the pathogenesis of sepsis, and was become a new target in the treatment of sepsis. Further research on the role of HMGB1 in the pathogenesis of sepsis is needed in the future, and the development of new drugs combined with HMGB1 will be used in the study of HMGB1 in animal experiments.

Objective To detect the expression of human epithelial growth factor receptor 2 (HER-2) in advanced lung adenocarcinoma with epithelial growth factor receptor (EGFR) mutation, and to explore the potential of HER-2 as a therapeutic target for drug resistance in patients with EGFR mutations. Methods HER-2 is commonly expressed in the advanced lung adenocarcinoma with EGFR mutations, mainly in the cell membrane. Results The overexpression rate of HER-2 protein in tissues of advanced lung adenocarcinoma with EGFR mutations was 33.3%(28/84). The overexpression rate of HER-2 protein in patients>50 years of age was 40.3%(27/67), which was significantly higher than that of patients ≤50 years of age [5.9 % (1/17)], the difference was statistically significant (χ2=7.227, P=0.007). The overexpression rate of HER-2 protein in patients with high pathological differentiation [44.4 % (8/18)] was higher than that in patients with poor pathological differentiation [30.3%(20/66)], but the difference was not statistically significant (χ2=1.273, P=0.259). The overexpression of HER-2 protein in patients with EGFR 21 exon mutation [40.5 % (17/42)] was significantly higher than that of EGFR19 exon mutation [25.0%(10/40)], but the difference was not statistical significance (χ2=2.222, P=0.136). Conclusions The overexpression rate of HER-2 protein in advanced lung adenocarcinoma patients with EGFR mutation is high, which is related to the age and tumor differentiation. HER-2 is expected to be a potential therapeutic target for drug resistance patients with EGFR mutations.

Objective To investigate the predictive values of P-selectin (P-sel),thrombus precursor protein (TpP) and D-dimer (D-D) in the early diagnosis of portal vein thrombosis (PVT) in patients submitted to splenectomy (or to devascularization).Methods The clinical data of 48 patients with portal hypertension (the PVT group,n=26; and the non-PVT group,n=22) who received operation in our hospital from 2009 to 2011 were retrospectively analyzed.Detecting the P-sel,TpP and D-D levels in the two groups of patients were done on preoperative day 1 and postoperative day 1,3,5,7,14.The SPSS software was used for statistic analysis.Results There were no significant differences on the preoperative day 1 levels of P-sel,TpP and D-D in the two groups (P＞0.05).The postoperative day1 levels of the three indicators in the PVT group were significantly higher than the non-PVT group (P＜0.05).Receiver operating characteristic (ROC) curves showed the area under curve (AUC) of P-sel was largest (0.893),followed by D-D,and TpP.The combined detection of the 3 indicators was highest,with the AUC up to 0.977.Conclusions Combined detection of P-sel,D-D and TpP levels were useful in the early diagnosis of PVT after splenectomy in patients with portal hypertension.