Objective To observe the effect of qixiantang decoction on asthma model mice and to explore its mechanism of phosphatase gene ( PTEN)-up-regulation. Methods A total of 28 healthy female BALB/c mice were divided into 4 groups according to the random number table ( n=7 ): normal control group, model control group, qixiantang decoction group, and dexamethasone group. The mice were sensitized with ovalbumin ( OVA) for asthma model. Qixiantang decoction group was treated with drug after OVA sensitization. Hematoxylin-eosin ( H-E) staining was applied to observe the pulmonary inflammation in mice, and periodic acid Schiff ( PAS) staining was used to examine airway mucus secretion. ELISA was used to detect the concentration of serum IgE. Real-time quantitative PCR was used to examine IL-13 and IL-5 gene expression changes in lung tissues of mice. Western blotting was used to detect the expression of PTEN and SIRT1 protein in lung tissues. Results The lung tissue inflammatory infiltration and mucus secretion in model control group were higher than normal control group (P<0. 01), and that in the qixiantang decoction group. The level of serum IgE in model control group [(6. 67 ± 2. 59) pg·mL-1)] was significantly higher than normal control group [(0.27 ± 0.05) pg·mL-1, P <0.01] ,and that in the qixiantang decoction group [(3.52 ±1.44) pg·mL-1,P<0.05]. The expression of PTEN and SIRT1 in lung tissue of model control group were significantly lower than normal control group, and that of qixiantang decoction group. The expression of IL-5 and IL-13 mRNA of qixiantang decoction group was significantly lower (P<0. 05). Conclusion Qixiantang decoction could significantly ameliorate inflammation in asthmatic mice by regulate IgE、IL-5、IL-13 expression, and might up-regulate PTEN expression via SIRT1 signal.

Objective To explore the in vitro antibacterial effect of tanreqing injection combined with cefuroxime sodium injection against staphylococcus aureus. Methods The MIC of tanreqing injection or cefuroxime sodium injection against staphylococcus aureus was detected by microamount dilution method.The antibacterial activity of tanreqing injection combined with cefuroxime sodium injection was determined by a chess board dilution method and assessed according to FIC index. Results The MIC of tanreqing injection and cefuroxime sodium injection against staphylococcus aureus was 1∶256 and 2 μg . mL-1 , respectively. While combined with each other, the MIC of tanreqing injection and cefuroxime sodium injection against staphylococcus aureus was 1∶4 096 and 0. 125 μg . mL-1 , respectively. The FIC index of tanreqing injection combined with cefuroxime sodium injection against staphylococcus aureus was 0. 125. Conclusion Tanreqing injection has a synergistic antibacterial effect against staphylococcus aureus when it was combined with cefuroxime sodium injection.

Objective To investigate the expression of connexin (Cx) 43, Cx26 gene in the thyroid tissue in patients with autoimmune thyroid diseases (AITD).Methods The thyroid tissue specimens of 76 cases were selected, including 30 cases of Graves disease (GD group), 30 cases of Hashimoto thyroiditis( HT group ), and 16 cases of normal tissues from thyroid adenoma (control group), which were all confirmed by pathologic diagnosis.The expression of Cx43 and Cx26 mRNA among three groups was determined by reverse transcriptase polymerase chain reaction (RT-PCR), and GAPDH,which was used as the internal control.Results The staining of Cx43 and Cx26 mRNA in all the cases from human thyroid tissue were strongly positive.The expression levels of Cx43,Cx26 mRNA in GD group were significantly higher than those in control group (0.9307 ± 0.0716 vs.0.5938 ± 0.0484 and 0.7371 ± 0.0463 vs.0.3431 ± 0.0399 ) (P ＜ 0.01 ).And the expression levels of Cx43,Cx26 mRNA in HT group were significantly lower than those in control group (0.3581 ±0.0458 vs.0.5938 ±0.0484 and 0.3150 ±0.0218 vs.0.3431 ±0.0399)(P＜0.01 or ＜0.05).Conclusion The Cx43 and Cx26 mRNA expression in the human thyroid tissue and a remarkable deferral expressions of Cx43 and Cx26 mRNA in three groups of AITD suggests that gap junction is associated with AITD.

Objective To explore the hierarchical chain management model of type 2 diabetes and determine its evaluation.Method Based on the hierarchical chain management of the three community health service institutions and Dahua hospital in Shanghai Xuhui district,215 cases of type 2 diabetes had been involved in the study.Results Compared with the baseline before management,lasting blood glucose (FBG),2 h postprandial glucose (2hPBG),glycosylated hemoglobin (HbA1c),low density lipoprotein cholesterol (LDL-C),systolic blood pressure (SBP) and diastolic blood pressure (DBP) of the diabetes after 12 months' management declined [(8.50 ±2.81) mmol/L,(11.09 ±4.01) mmol/L,(8.56 ±2.41)% ,(3.31 ± 1.06) mmol/L,(139.06 ±20.68) mm Hg (1 mm Hg = 0.133 kPa),(78.20 ± 12.11) mm Hg vs.(7.41 ±2.04) mmol/L,(9.03 ±2.46) mmol/L,(7.34 ± 1.59)% ,(3.00 ± 1.06) mmol/L,(135.48 ± 17.82) mm Hg,(77.27 ±11.83) mm Hg],and the differences were statistically significant(P＜0.01 );control rate of FBG,2hPBG,HbA1c,LDLC,SBP,DBP had improved significantly [19.5% (42/215),20.9% (45/215),24.7%(53/215),20.0%(43/215),27.4%(59/215),30.2%(65/215) vs.50.7%(109/215),53.0% (114/215),54.0%(ll6/215),42.3%(91/215),47.0%(101/215),45.6%(98/215)](P＜0.01).Conclusion Primary and secondary-care hospital based hierarchical chain management model is valid and can be implemented for type 2 diabetes.

Objective To investigate the relationship and mechanism of the heme oxygenase-1 expression in peripheral blood mononuclear cells (PBMC) and the oxidative stress in diabetic nephropathy. Methods Two groups of diabetic patients with or without diabetic nephropathy and a normal control group were enrolled in this study. Fasting blood glucose (FBG), HbA1c, serum MDA level, ROS level, HO-1 mRNA level and HO-1 protein expression in PBMC were determined. Results In control group, diabetic group and diabetic nephropathy group , the MDA levels significantly increased[(14.23±5.07)nmol/ml vs (24.90±7.12)nmol/ml vs (43.83±16.97)nmol/ml](F＝37.022,P＜0.01), the ROS levels significantly increased (113.18±58.59 vs 364.54±88.67 vs 524.35±162.51)(F＝68.369,P＜0.01) and the HO-1 protein expression also increased significantly (22.84±9.98 vs 36.72±15.85 vs 58.1±15.93)(F＝31.302,P＜0.01). There was a positive correlation among the HO-1 mRNA, protein expression and MDA level(r＝0.407,0.429,P＜0.05). Conclusions There existed a severer oxidative stress condition in patients with diabetic nephropathy compared with the patients without diabetic nephropathy. HO-1 could be a potential pathway to ameliorate oxidative stress in diabetic kidney disease patients.

Objective To explore the hierarchical chain management model in blood glucose control and its influence factors in patients with diabetes mellitus.Methods Health management database of diabetic patients was established in 2007 and managed by hierarchical chain management.The number of the patients reached to 1010 till 2011.The blood glucose control of diabetic patients was analyzed and its influence factors were analyzed by multivariate unconditional Logistic regression method.Results The concentration of glycosylated hemoglobin( HbA1c ) of 1010 patients with type 2 diabetes was (8.21 ±:2.70)％.Four hundred and eighty-seven cases (48.22％) reached the blood glucose standard,303 cases (30.00％)reached the blood pressure standard,245 cases (24.26％) reached the blood lipids standard,and 76 cases (7.52％) reached all three standards.Multivariate analysis showed that occupation (OR =2.521,95％ CI:1.871 - 3.397),education level (OR =1.890,95％ CI:1.642 - 2.174),disease course (OR =1.035,95％CI:1.016 -1.055),systolic pressure (OR =1.016,95％ CI:1.007 -1.025) and triglyceride (OR =1.204,95％CI:1.063 - 1.365) were the risk factors of blood glucose control (P ＜0.01).Conclusions Hierarchical chain management model is helpful for the blood glucose control in patients with type 2 diabetes.The comprehensive control and treatment of type 2 diabetes should be taken combined with related risk factors,such as blood pressure,blood lipids and diabetes disease course.

Objective To evaluate the potential use of a probe melting analysis (PMA) assay in detecting the embB mutations which confer resistance against ethambutol in Mycobacterium tuberculosis. Methods The analysis sensitivity and specificity of PMA were investigated by detecting a serially diluted H37 Rv DNA and a reference panel from National Institute for the Control of Pharmaceutical and Biological Product. Six hundred and thirteen sputum samples were collected from the Xiamen Center for Disease Control and Prevention, Xiamen First Hospital and Center for Zhangzhou Disease Control and Prevention from September 2009 to April 2010. The PMA assay was then evaluated by detecting 613 clinical isolates and the results were compared with the sequencing results. Results The PMA assay could specifically detect Mycobacterium tuberculosis and had a limit of detection of 3 copies per reaction. The assay results with 613 clinical isolates showed that PMA gave a 100% concordance with sequencing in the 583 qualified samples, among which 34 were mutations at embB 306,23 at embB 378-380, 3 at embB 406 and 3 at embB 497. Conclusions PMA assay is a sensitive and specific method enabling efficient detection of common embB mutations causing ethambutol-resistance. The rapidness of this method together with its reliability would facilitate its use in routine testing.

Objective To analysis the influence factors of standardization in the hierarchical chain management of type 2 diabetes and to enhance the hierarchical chain management of type 2 diabetes.Methods ( 1 ) Six hundred and ninty patients with type 2 diabetes completed 1 years management were divided into well-controlled glycosylated hemoglobin ( HbAlc ) group (＜7.0％ ) and bad-controlled glycosylated hemoglobin (HbAlc) group ( ≥ 7.0％ ).The conditions of diet,physical activity,medication,self-blood sugar monitoring and participation in health seminars were investigated and analyzed.(2) The patients were divided into standardized management group and not standardized management group.Their age,sex,educational background,occupation,monthly income per person,medical security,the course,cognition for glycuresis,two-way transfer,and chronic complications were investigated and statistically analyzed.Results ( 1 ) The proportions of physical activity (70.1％ vs 54.2％,x2=6.163,P=0.018),self-blood sugar monitoring(60.4％ vs 43.8％,x2=6.268,P=0.016) and participation in health seminars (56.0％ vs 41.7％,x2=4.577,P=0.045) in the well-controlled HbAlc group were significantly higher than those in the bad-controlled HbAlc group.(2) Their age [(61.08 ±10.04) years old vs ( 57.75 ± 9.89 ) years old,t=2.539,P=0.012],educational background ( ratio of low educational attainment:8.3 ％ vs 17.2％,x2=6.426,P=0.041 ),medical security (own expense ratios:4.6％ vs 11.5％,x2=3.543,P=0.048 ),awareness of diabetes ( ratio of poor awareness of diabetes:19.4％ vs 41.0％,x2=17.518,P=0.000 ),two-way transfer ( ratio of not transfer treatment:4.6％ vs 14.8％,x2=7.662,P=0.022) and chronic complications ( ratio of chronic complication:41.7 ％ vs 26.2％,x2=6.130,P=0.017) were significantly different between the standardized management group and not standardized management group.(3) Logistic regression analyses indicated that the age ( OR=0.954,P=0.006),monthly income per person ( OR=4.101,P=0.018 ),medical security ( OR=7.617,P=0.003 ),cognition for glycuresis ( OR=0.030,P=0.000),two-way transfer ( OR=9.079,P=0.000) and chronic complications ( OR=0.456,P=0.031 ) were the risk factors of standardized management.Conclusion We should focus on the impact factors affecting the standardized management of patients including age,monthly income per person,medical security,awareness of diabetes,ratio of not transfer treatment,positive strategies for chronic complications,improve the hierarchical chain management of type 2 diabetes,and then make the diabetic patients to early participate in standardization management of diabetes mellitus and delay the appearance of complications.

OBJECTIVETo prepare streptavidin-tagged human interferon-inducible T cell alpha chemoattractant bifunctional fusion proteins (SA/hI-TAC) and evaluate its biological activity.

METHODSpET24a-SA-hI-TAC/pET21a-hI-TAC-SA plasmids were constructed and expressed in BL21. SA-hI-TAC and hI-TAC-SA fusion proteins were purified by Ni-NTA affinity chromatography, refolded by dialysis and identified by Western blotting. The bifunctionality of the fusion proteins (biotin-binding function and hI-TAC activity) was analyzed by flow cytometry and lymphocyte chemotaxis experiment, respectively.

RESULTSSA-hI-TAC/hI-TAC-SA fusion proteins were expressed at about 12% and 25% of the total bacterial protein, respectively. The two fusion proteins had a purity of about 85% and 90% after purification, and their purity reached 98% after purification with S-100 gel filtration chromatography. Both of the fusion proteins were efficiently immobilized on the surface of biotinylated mouse bladder cancer MB49 cells (91.3% for SA-hI-TAC and 98.8% for hI-TAC-SA). SA/hI-TAC induced lymphocyte chemotaxis in a dose-dependent manner, and hI-TAC-SA showed a stronger chemotactic effect than SA-hI-TAC.

CONCLUSIONSWe successfully obtained SA/hI-TAC bifunctional fusion proteins, which may potentially be used in local treatment of tumor and as a tumor vaccine.

Animals ; Biotinylation ; Blotting, Western ; Cancer Vaccines ; Cell Line, Tumor ; Chemokine CXCL11 ; chemistry ; Chromatography, Affinity ; Humans ; Interferons ; chemistry ; Mice ; Plasmids ; Recombinant Fusion Proteins ; biosynthesis ; chemistry ; Streptavidin

Equine interferon-gamma (eIFN-gamma) expressed both in E. coli and baculovirus were evaluated for antiviral activity against recombinant Vesicular Stomatits Virus expressing green fluorescence protein (rVSV-GFP) in EFK-78 cells. The assays were conducted in 96-well plate. Virus infectivity was measured by quantifying GFP-positive cells, instead of quantifying the CPE reduction. Prior to infection of EFK-78 cells with rVSV-GFP, the cells were incubated with eIFN-gamma. The GFP expression in the EFK-78 cells dramatically decreased in the cells treated with eIFN-gamma in a dose-dependent manner, comparing with the mock-treated cells. The titers of antiviral activity were 1 x 10(3) AU/mL and 1 x 10(5) AU/mL of eIFN-gamma expressed from E. coli and baculovirus, respectively. The antiviral activities of the recombinant eIFN-gamma were highly efficient and specific, as it was blocked by mAbs against eIFN-gamma.
Animals ; Antibodies, Monoclonal ; immunology ; Antiviral Agents ; metabolism ; pharmacology ; Baculoviridae ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Green Fluorescent Proteins ; metabolism ; Horses ; Interferon-gamma ; biosynthesis ; genetics ; pharmacology ; Recombinant Proteins ; Vesicular stomatitis Indiana virus ; drug effects ; metabolism