Objective:To observe the effect of Radix Paeoniae Bubra(RPB)on NADPH oxidase p22phox,monocyte chemotactic protein-1(MCP-1)mRNA expression and neointimal hyperplasia after carotid artery balloon injury in cholesterol-fed rabbits.Methods:The rabbit model of carotid balloon injury was established adopting Clowes method,and treated with extract of RPB.Component of neointima and expression of proliferating cell nuclear antigen(PCNA)and macrophage was determined by immunochemical stain.The collagen of typeⅠwas detected by special staining for blood vessels and the area of neointima was measured by image assay system.Expression of NADPH oxidase p22phox mRNA,MCP-1 mRNA was detected by in situ hybridization and transcription-polymerase chain reaction.Results:RPB can attenuate the neointima area and proliferation of collagen typeⅠinduced by balloon-injury,remarkable prevent the formation of restenosi,and down-regulate expression of NADPH oxidase p22 and MCP-1mRNA significantly and decrease the degree of macrophages infiltration especially in vessel wall of injuring carotid artery.Conclusions:RPB inhibited NADPH oxidase P22Phox and MCP-1 mRNA expression,and modestly reduced neointimal hyperplasia,which might be partly attributed to its antioxidant and inflammatory effects.

AIM:To establish an HPLC method for determining the contents of caffeic acid,quercetin and campherenol in Hedyotis diffusa Willd. METHODS:The samples were separated on an Alltima C 18 (250 mm? 4.6 mm,5 ?m) column with the mobile phase of MeOH(A)-0.5% glacial acetic acid solution;gradient elution(0～15 min,30%～60% A;15～30 min,60%～60% A).Flow rate was 1.0 mL/min. The detection wavelength was set at 350 nm.Column temperature was at 30 ℃. RESULTS:The contents of caffeic acid,quercetin and campherenol were 14.218～23.695 ?g/g,9.919～25.564 ?g/g and 6.229～18.160 ?g/g in Hedyotis diffusa Willd from different sources. The linear range of caffeic acid was 0.005 0～0.200 0 ?g(r=0.999 9),the average recovery was 102.35%,RSD was 2.31%(n=6);The linear range of quercetin was 0.006 2～0.244 0 ?g(r=0.999 9),the average recovery was 101.84%,RSD was 1.79%(n=6);The linear range of campherenol was 0.007 8～ 0.310 6 ?g(r=0.999 9),the average recovery was 99.04%,RSD was 2.90%(n=6). CONCLUSION:The method for quantifying of caffeic acid,quercetin and campherenol in Hedyotis diffusa Willd is accurate and reliable.

Objective To investigate the relationship between OHSS and FSHR gene polymorphism. Methods Two hundred and two women were enrolled in this study. The FSHR gene polymorphisms at position 307 and 680 were detected. Results The distribution of the allele frequency and genotype frequency of the position 680 in FSHR gene were significantly different between women with OHSS or not. No significant differences of the position 307 in FSHR gene were observed. Conclusion Themutation of Asn680Ser in FSHR might be closely related with OHSS.

Objective To evaluate the performance of enzyme linked immunosorbent assay (ELISA) kit in detection of Hepatitis B virus(HBV) markers by using improved and optimized method ,so as to provide a practical and feasible method and reagents for clinical laboratory .Methods ELISA test was used for the detection of HBV markers .The gradient dilution method was used to e‐valuate the lower limit .The verifiation of cut off value was carried out based on clinical and laboratory standards institute (CLSI) EP12‐A2 document .Samples with cut off values were collected to evaluate the precision ,including repeatability and intermediate precision .The coincidence rates were counted through comparing the results of ELISA with those of external quality assessment and those detected by using Abbott i4000SR chemiluminescence instrument .Results The lower detection limit of HBsAg ,HBsAb , HBeAg ,HBeAb and HBcAb were 0 .2 IU/mL ,20 mIU/mL ,1 NCU/mL ,0 .75 NCU/mL and 0 .05 NCU/mL respectively .The cut‐off value(C50 )± 20% concentration included the concentration range between C5 and C95 .The within‐run coefficient of variation (CV)≤15% ,in sandwich method the between‐run CV≤25% ,in competition method the between‐run CV≤35% .The positive and negative coincidence rates in accuracy and comparing with i 4000SR all were more than 95% and all κ>0 .75 .Conclusion ELISA tests for HBV markers in our laboratory could meet the requirements of the detection performance and clinical needs .

Based on the demand of nasal drug delivery high drug loadings, using the unique phase transfer of solute, integrating the phospholipid complex preparation and submicron emulsion molding process of Scutellariae Radix extract, the study obtained the preparation of the high drug loadings submicron emulsion of Scutellariae Radix extract. In the study of drug solution dispersion method, the uniformity of drug dispersed as the evaluation index, the traditional mixing method, grinding, homogenate and solute phase transfer technology were investigated, and the solute phase transfer technology was adopted in the last. With the adoption of new technology, the drug loading capacity reached 1.33% (phospholipid complex was 4%). The drug loading capacity was improved significantly. The transfer of solute method and timing were studied as follows,join the oil phase when the volume of phospholipid complex anhydrous ethanol solution remaining 30%, the solute phase transfer was completed with the continued recycling of anhydrous ethanol. After drug dissolved away to oil phase, the preparation technology of colostrum was determined with the evaluation index of emulsion droplet form. The particle size of submicron emulsion, PDI and stability parameters were used as evaluation index, orthogonal methodology were adopted to optimize the submicron emulsion ingredient and main influential factors of high pressure homogenization technology. The optimized preparation technology of Scutellariae Radix extract nasal submicron emulsion is practical and stable.
Administration, Intranasal ; Emulsions ; Plant Extracts ; Technology, Pharmaceutical ; methods

OBJECTIVETo investigate the effects of chronic stress on the spatial learning-memory and the role of glial cell line-derived neurotrophic factor (GDNF) of prefrontal cortex (PFC) and hippocampus (HP) in different age mice.

METHODSThe chronic stress model mice in 21 days with multiple chronic unpredictable stressors were applied. The spontaneous behavior and spatial learning-memory ability of mice were tested, using Open field and Morris water maze task, and the expression of GDNF in HP and PFC were detected by immunohistochemical method.

RESULTSCompared with young mice, the spontaneous behaviors were significantly decreased and the spatial learning-memory function were significantly decreased (P < 0.05, P < 0.01) in aged mice. The GDNF expression in the CA3, DG of HP and PFC were significantly reduced in aged mice (P < 0.05, P < 0.01). After chronic stress, the spontaneous behaviors were remarkably decreased and the ability of spatial learning-memory of the stress group mice were significantly decreased (P < 0.05, P < 0.01) compared with those of the control group mice. The expression of GDNF in HP and PFC were remarkably reduced (P < 0.05, P < 0.01) in stress group mice. The aged stress mice had more serious changes after chronic stress.

CONCLUSIONThe brain aging and chronic stress in mice causes behavioral changes and the damage of spatial learning-memory function, and which may be nearly related to the expression of GDNF in HP and PFC.

Aging ; Animals ; Cerebral Cortex ; metabolism ; Cognition Disorders ; metabolism ; Female ; Glial Cell Line-Derived Neurotrophic Factor ; metabolism ; Hippocampus ; metabolism ; Male ; Maze Learning ; Mice ; Mice, Inbred Strains ; Stress, Physiological

OBJECTIVETo investigate the expression of transcription factors (TF) T-bet and GATA-3 mRNA in peripheral blood mononuclear cells and its correlation with immune status in esophageal cancer patients.

METHODSSixty patients were divided into two groups according to the clinical data: group A consisting of stage I and II, group B including stage III and IV. The gene expression of T-bet and GATA-3 in 60 esophageal cancer patients and 30 healthy controls was detected by reverse transcription-polymerase chain reaction (RT-PCR). The expression of IFN-gamma and IL-4 was measured by enzyme linked immunosorbent assay (ELISA).

RESULTSExpression of T-bet mRNA in esophageal cancer patients (stage I and II: 0.27 +/- 0.05 ng/L, stage III and IV: 0.12 +/- 0.02 ng/L) was significantly lower than that in the healthy controls (1.35 +/- 0.14 ng/L), but the expression of GATA-3 mRNA in esophageal cancer patients (stage I and II: 0.45 +/- 0.06, stage III and IV: 0.55 +/- 0.03) was significantly higher than that in the healthy controls (0.09 +/- 0.10). The plasma level of Th1 cytokine IFN-gamma in the patients [stage I and II: (12.12 +/- 1.48) ng/L, stage III and IV: (8.44 +/- 0.90) ng/L] was significantly lower than that in the healthy controls, while the level of Th2 cytokine IL-4 in the patients [stage I and II: (18.64 +/- 0.77) ng/L, stage III and IV: (25.28 +/- 2.02) ng/L] was significantly higher than that in the healthy controls. However, neither in the expression of T-bet and GATA-3, nor in the plasma level of IFN-gamma and IL-4, showed a significant difference between group A and B.

CONCLUSIONIn the peripheral blood of esophageal cancer patients, the expression of T-bet decreased, while GATA-3 increased, Th1/Th2 balance is broken, and the Th2 is dominant. T-bet and GATA-3 play a part role in the regulation of Th1/Th2 balance.

Adult ; Aged ; Aged, 80 and over ; Enzyme-Linked Immunosorbent Assay ; Esophageal Neoplasms ; immunology ; metabolism ; pathology ; Female ; GATA3 Transcription Factor ; genetics ; metabolism ; Humans ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; Neoplasm Staging ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; T-Box Domain Proteins ; genetics ; metabolism ; Th1 Cells ; metabolism ; Th2 Cells ; metabolism

Objective To study the effect of TLR4 activation with LPS on BMP9-induced osteogenic differentiation of immortalized mouse embryonic fibroblasts ( iMEFs).Methods The activation of TLR4/NF-κB signaling path-way was detected by ICC.iMEFs were treated with LPS,BAY11-7082,Adnovirus GFP and BMP9.The early osteo-genic differentiation capability of iMEFs was detected by ALP staining and quantitative assay .The later osteogenic differentiation capability was detected by alizarin red S staining .The expression of later osteogenic differentiation marker gene OCN and OPN were detected by PCR and Western blot .The change of p-Smad1/5/8 was detected by Western blot.The expression of Runx2 and Dlx5 were detected by PCR and Western blot .Results LPS can effec-tively stimulate TLR4/NF-κB signaling pathway .TLR4 activation inhibited BMP 9-induced osteogenic differentiation . BMP9-induced osteogenic differentiation related gene and Smad 1/5/8 signaling activation were inhibited by TLR4 activation .The inhibition effect was partly reversed by BAY 11-7082 ( P<0.05 ) .Conclusions TLR4 activation with LPS can inhibit BMP9-induced osteogenic differentiation of iMEFs cells via NF-κB signaling pathway .

The effect of C-terminal region residues on the substrate specificity of a novel cyclic imide hydrolase (CIH), a recombinant cyclic imide hydrolase (CIH293), and its mutants deleted or substituted at C-terminus (CIH291, CIH290, KK292-293EE) was reported. The substrate specificity and kinetic parameters of the mutants were analyzed by both the spectrophotometric assay and high-performance liquid chromatography. Results show that the substrate specificity of mutants was not obviously changed, but slightly low for the affinity between the substrate and enzyme, compared with the wild-type enzyme, CIH293. In conclusion, the last three residues of CIH293 play an important role for the enzyme activity.

Objective:To prepare mesalazineβ-cyclodextrin inclusion compound and observe its properties. Methods:The struc-ture was characterized by UV, IR and DSC. The inclusion process of mesalazine andβ-cyclodextrin was simulated using molecular doc-king technique. The solubility and dissolution of inclusion compound were determined. Results:UV, IR and DSC were used to deter-mine the formation of inclusion complex. The inclusion ratio was 1: 1 and the phase solubility diagram was AL type. The solubility of inclusion compound in deionized water (25 ℃, pH 7. 0) was 7. 8 mg·ml-1 . The preparation of mesalazine β-cyclodextrin inclusion compound could significantly increase the dissolution rate and solubility of mesalazine. Conclusion:The prepared mesalazineβ-cyclo-dextrin inclusion compound can obviously improve the poor water solubility and dissolution of mesalazine.