Objective To observe the effects of intensive insulin therapy on IL-10 level and NF-ΚB activity in peripheral blood mononuclear cells in patients undergoing cardiopulmonary bypass.Methods The non-diabetic patients undergoing cardiopulmonary bypass in our department were selected and assigned to intensive therapy group (group A,n=40) and received strict glycemic control after the initiation of surgery.And those who undergoing cardiac surgery but without strict glycemic control were assigned to routine therapy group (group B,n=40) as controls.The blood glucose in group A was maintained at 4.4～8.3mmol/L,whereas the glucose in group B was below 11.lmmol/L.The concentration of serum IL-10 and NF-ΚB activity in peripheral blood mononuclear cells was measured at different time points.Results There were no significant differences in general data between two groups.The concentration of IL-10 in group B was significantly lower than that in group A(P<0.05).compared with group B,strict glycemic control markedly suppressed NF-KB activation (P<0.05).Conclusion Intensive insulin therapy could reduce the activity of NF-ΚB and then reduce the expression of IL-10.Strict glycemic control could significantly mitigate the systemic inflammatory response.

Objective To study ultrasonic parameters characteristics of lactation mastitis,breast cancer by using color doppler flow imaging(CDFI)and ultrasonic elastography(UE),and to explore its clinical value.Methods Retrospective analysis was conducted in the clinical data of 85 female patients (a total of 85 breast lesions).All patients were confirmed by pathology,and according to the results of pathology,they were divided into non lactation mastitis group(group NLM,a total of 28 cases),breast cancer group(group Ca,a total of 57 cases).All patients accepted CDFI and UE before treatment,and compared ultrasound parameters between the two groups.Results The proportions of class 0~I(71.43%)and RI<0.7(82.14%)in group NLM were significantly higher than those in group Ca,while the proportions of classⅡ ~Ⅲ(28.57%)and RI≥0.7 (17.86%)were significantly lower than 75.44%,84.21% in group Ca(χ2 =17.185,35.217,all P<0.05).The UE ratings(1.75 ±0.97)and the strain rate ratio (1.64 ±0.83)in NLMgroup were lower than (4.19 ±0.74),(5.03 ±1.08)in group Ca(t=-12.873,-14.623,all P<0.05).The accuracy of CDFI +UE(89.41%)was higher than the accuracy of UE(76.47%)or the accuracy of CDFI(67.06%)(χ2 =12.337,P<0.05).Conclusion CDFI,UE have a certain diagnosis ability for non lactation mastitis and breast cancer,combining both can obtain better diagnostic value,which is worthy of clinical application.

Objective:To prepare and purify siRNA targeting a proliferation-inducing ligand targeted(APRIL-siRNA),so as to provxde a basis for studying the role of APRIL in human pancreatic cancer.Methods:pET-22b-APRIL was constructed to express APRIL dsRNA of human pancreatic cancer cell line CFPAC-1 in E.coli and the product was purified by chromatography using CF-11 column.APRIL dsRNA was digested by RNaseⅢto prepare APRIL siRNA,then the reaction mixture was loaded onto a DEAE ion exchange chromatography to remove RNaseⅢfrom oligonucleotides,and size exclusion chromatography was used to purify 21 bp siRNA.The purified APRIL siRNA was used to transfect Chinese hamster ovary(CHO)cells and the expression of APRIL in CHO cells was observed under fluorescence microscope Results:APRIL dsRNA was successfully expressed in E.coli after IPTG induction and was purified by CF-11 column.dsRNA was hydrolyzed with RNaseⅢand was purified by DEAE ion exchange chromatography and size exclusion chromatography.15% nondenaturing PAGE and 12% SDS- PAGE confirmed that RNaseⅢwas removed from oligonucleotides and 21 bp siRNA was purified with size exclusion chromatography.It was also found that APRIL siRNA obviously depressed APRIL expression in CHO cells.Conclusion:We have successfully constructed APRIL siRNA targeting APRIL gene of CFPAC-1 cells with in vitro transcription,which provides a basis for knock-down of APRIL gene in CFPAC-1 cells.

Objective To analyze the ischemia degree of initial electrocardiogram in the patients of acute ST-segment elevation myocardial infarction (STEMI) and investigate its value in the predicting cardiogenic shock (CS) after percutaneous coronary intervention (PCI).Methods Three hundred and ninety patients with STEMI from the onset of symptoms to admission within 12 h were divided into two groups based on grade Ⅱ ischemia (group A,248 cases) or grade Ⅲ ischemia (group B,142 cases) in the initial electrocardiogram.Clinical data,TIMI risk score,ST-segment resolution (STR),CS and cardiovascular events (hospital mortality,ventricular arrhythmias,reinfarction) were recorded in all patients.Results The gender,time from onset to balloon opening,smoking,hypertension,type 2 diabetes,hyperlipidemia,stroke,postoperative TIMI flow 3 grade,coronary artery lesions and lesions in the left main stem between two groups was not statistically significant (P ＞0.05).In group B,the rate of ST segment resolution ＞ 50％ was significantly lower than that in group A [53.2％ (132/248) vs.29.6％ (42/142)] (P ＜ 0.01).The incidence of CS,in-hospital death,malignant ventricular arrhythmias in group B was higher than that in group A,and the difference was statistically significant (P ＜ 0.05).The age,left ventricular ejection fraction,TIMI risk score ＞ 3 points,Killip grade ＞ 1 grade,anterior myocardial infarction between two groups was statistically significant (P ＜ 0.05 or ＜ 0.01).The indicators that were statistically significant in the univariate analysis were included into Logistic regression model and analyzed,with CS-related factors as independent variables and CS as the dependent variable and found that age (P =0.008),Killip class ＞ 1 grade (P =0.049),ST segment resolution rate (P =0.008) and grade Ⅲ ischemia (P =0.001) as independent predictors of CS after PCI.Conclusions Grade Ⅲ ischemia is an independent predictor of CS after PCI in STEMI patients.And it has predictive value for hospital mortality and ventricular arrhythmias.

Objective To assess the value of hemofiltration in the prevention of contrast induced nephropathy(CIN) post percutaneous coronary intervention(PCI) in severe chronic kidney disease stages(CKD) patients. Methods We evaluated 30 CKD patients who underwent PCI followed by bedside hemofiltration. We measured serum creatinine levels before PCI, 24 hours, 72 hours, and one week after PCI, and calculated creatinine clearance(CrCl) according to Cockcroft and Gault equation.We observed the incidence of CIN, and the short term clinical efficacy of hemofiltralion. Results The average age of the 30 patients was (72.87 ± 8.71) years old, with 21 (70%) male patients. The stages of CKD among the patients included CKD 3 (3 patients, 10%), CKD 4 (20 patients, 66.7%) and CKD 5 (7 patients, 23.3%). The average duration of hemofiltration was (7.5±4.1) hours. Serum creatinine before PCI, 24 hours, 72 hours and 1 week after PCI was (498.7±143.7)μmol/L, (353.2±128.0)μmol/L, (450.0±132.2)μmol/L, (488.0±145.7)μmol/L respectively,and CrCl was (20.3±10.2) ml/min, (36.5±14.3) ml/min, (28.3±10.4) ml/min, (21.0±10.3) ml/min respectively. There was no CIN. Mean follow-up was (3.1 ± 2.6) months. There was no new cardiovascular events, and no new patients need to rely on long term hemodialysis. Conclusions For patients with severe renal insufficiency, hemofiltration may reduce the incidence of CIN, It is an alternative preventive measures to prevent CIN.

Objective To investigate the expression of normal epithelial cell-specific-1(NES1) gene in normal gastric epithelial cells and different gastric cancer cell lines and the effects of 5-aza-2-de- oxycytidine(5-aza-dC)on the expression of NES1 gene.Methods Expression of NES1 mRNA in five gastric cancer cell lines(MKN-28,SGC-7901,AGS,MKN-45 and HGC-27)and normal human gastric epithelial cells were detected by real-time PCR.After treatment with 5-aza-dC,a DNA methyltransferase inhibitor,the expression of NES1 mRNA in gastric cancer cell lines was detected by real-time PCR. DNA methylation status of NES1 gene was assayed by methylation-specific PCR(MSP).Results The expression of NES1 mRNA was decreased in all gastric cancer cell lines.A strong correlation between ex on 3 hypermethylation and loss of NES1 mRNA expression in gastric cancer cell lines was noted.5-aza dC treatment of NES1-nonexpressing tumor cell lines resulted in a dose-dependent induction(SGC-7901, MKN-45,MKN-28 and AGS)and increase (HGC-27) of NES1mRNA expression in gastric cancer cells. Conclusions The study suggested that hypermethylation was a responsible factor for tumor-specific loss of NES1 gene expression in gastric cancer cells.Treatment of gastric cancer cell lines with a demethylating agent led to reexpression of NES1 suggesting an important role of hypermethylation in loss of NES1 gene expression.

Objective To evaluate efficacy and safety of fixed combination of nitrendipine and atenolol in treatment for patients with mild to moderate essential hypertension and their optimal dosage matching.Methods Totally,275 patients with essential hypertension were selcted from seven hospitals in Shanghai,Nanjing and Suzhou,China and randomized into five groups with same proportional probability in a double-blind,double-dummy,parallel active-controlled,multi-center clinical trial,receiving fixed combination of nitrendipine and atenolol at three different dosage matching (nitrendipine/atenolol 5/12.5 mg,5/10 mg,5/7.5 mg for groups 1,2 and 3),and nitrendipine (10 mg for group 4) or atenolol (25 mg for group 5),respectively for eight weeks.Results Mean reduction of diastolic blood pressure (DBP)was (17±7) mm Hg,(18±9) mm Hg and (17±7) mm Hg for groupl,2 and 3,respectively from the baseline,significantly greater than that in groups 4 and 5[(13±7) mm Hg and (12±6) mm Hg,respectively].Mean reduction of systolic blood pressure (SBP) was (21 ±11)mm Hg,(24±12) mm Hg,(23±11) mm Hg,(19±13) mm Hg and (18±9) mm Hg,respectively for the five groups from the baseline,and the reduction in group 2 was significantly greater than that in group 5,with an overall efficacy of 94.4%,98.1% and 88.2% for groups 1,2 and 3,respectively,all statistically higher than that in group 5 (71.4%) with P＜0.01,eight weeks after treatment.The ratio of patients with increased dose of antihypertensive agents in week 5 was lower in group 2 than that in the other four groups,with mild adverse reaction only,no obvious change in laboratory biochemical examinations,and no needs in special management.Conclusions Fixed combination of atenolol and nitrendipine with an optimal doses of 5 mg and 10 mg respepctively was effective and safe for mild and moderate hypertension with good tolerance.

Objective To establish a time resolved fluoroimmunoassay (TRFIA) method for detecting Glypican 3 (GPC3) and to explore the diagnostic value of serum GPC3 for hepatic carcinoma (HCC). Methods Microplate coated with anti-GPC3 monoclonal antibody 7C8 and GP9 labeled with Eu3+ were used to establish TRFIA kit. The serum concentrations of GPC3 in 41 HCC patients and 44 chronic hepatitis (CH) patients were quantitatively analyzed. AFP was detected by with lowest limit of 2.06 μg/L. The CV of inter and intra assay were 12.25% and 12.91%, respectively. The average serum concentration of GPC3 in HCC patients was (86.68±110.39) μg/L (median: 56.98 μg/L). But in CH patients it was only (14.77±29.48) μg/L, which was significantly lower than that in HCC (Wilcoxon W=1335.00, Z=-4.99, P<0.001). With diagnostic cut-off value set at 42.94 μg/L, the diagnostic sensitivity and specificity of TRFIA GPC3 for HCC were 58.5% (24/41) and 95.5%(42/44) respectively. The diagnostic sensitivity of AFP was 46.3% (19/41) in 41 HCC patients, and was raised to 78.0% (32/41) when combined with GPC3. Conclusions Serum GPC3 assay by TRFIA is established and it could increase the diagnostic sensitivity for HCC when combined with AFP.

OBJECTIVETo develop an ELISA-based method for analyzing biologic activities of HIV-1 integrase and for high throughput screening of integrase inhibitors.

METHODSAfter expression, renaturation and purification of integrase, the bioactivity of integrase and the inhibition of luffin-a were evaluated with an in vitro assay based on biotin-avidin EILSA and chemiluminescent substrates.

RESULT(1) The specific activity of the purified integrase was 54.92 units/mg of protein. (2)IC(50) (concentration causing 50% inhibition of integrase) of luffin-a was (0.63 +/- 0.026) micromol/L.

CONCLUSIONThe non-radioactive assay can be used for analysis of bioactivities and high throughput screening of inhibitors of HIV-1 integrase.

Catalysis ; drug effects ; Dose-Response Relationship, Drug ; Enzyme Inhibitors ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; methods ; HIV Integrase ; chemistry ; metabolism ; Humans ; Kinetics ; Luminescent Measurements ; Ribosome Inactivating Proteins, Type 1 ; pharmacology ; Substrate Specificity

The combination effect of high hydrostatic pressure and moderate heat on the inactivation kinetics of Bacillus coagulans spore in phosphate buffer and UHT(Ultra High Temperature)whole milk was investigated.The pressure come-up time and corresponding log-reduction of spore inactivation were considered during pressure-thermal treatment.Bacillus coagulans spore had a much higher resistance to pressure in UHT whole milk than in phosphate buffer.Survival data were modeled using the linear,Weibull and log-logistic models to obtain relevant kinetic parameters.The tailing phenomenon occurred in all survival curves,indicating the linear model was not adequate for describing these curves.The log-logistic model produced best fits to survival curves,following by Weibull model.