Objective To isolate and identify the antitumor bioactive components from the fermentation of the fungus Penicillium auratiogriseum Sp-19 derived from sponge Mycale plumose.Methods The antitumor components were isolated by bioassay-guided fractionation and using solvent extraction,silica gel column chromatography and preparative HPLC.Their structures were established by physicochemical properties and spectral analyses.The cytotoxicities of compounds were evaluated by SRB method.Results and Conclusion Five alkaloid compounds were isolated and identified as fructigenines A(1),deacetyl fructigenines A(2),fructigenines B(3),1,4-benzodiazepine-2,5-diones(4) and cyclopenin(5),respectively.Compound 4 and 5 showed cytotoxicity at the concentration of 3 ?g?mL~(-1)against tsFT210 cell line.

OBJECTIVETo evaluate the effect of concentrated growth factor extract (CGFe) on the proliferation and differentiation of MC3T3-E1 osteoblasts attached to sandblasted and acid etched titanium surfaces.

METHODSTrials were divided into experimental and control groups. The experimental group used a-MEM that contained CGFe (10% FBS), whereas the control group only used a-MEM (10% FBS). MTT assay was employed to detect the number of osteoblasts on the first, third, and fifth days. Alkaline phosphatase (ALP) activity and scanning electron microscope (SEM) were used to detect the osteoblast differentiations on the third and fifth days and to observe the osteoblast extensions on titanium surfaces for 12 h, respectively. Meanwhile, the levels of the osteogenetic biomarkers Runt-related transcription factor-2 (Runx2) and Osterix (Osx) on the third and seventh days were quantified via real-time polymerase chain reaction (PCR).

RESULTSMTT assay indicated that on the first, third, and fifth days, the absorbance in the experimental group significantly increased than that in the control group (P < 0.05). ALP activity: on the third and fifth days, the absorbance of the experimental group was significantly higher than that of the control group (P < 0.05). SEM: at 12 h, the extension of the experimental group cells was larger than that of the control group. Real-time PCR: given the standardization in the group, the gene expression level of the control group on the third day was 1, and the Runx2 and Osx gene expressions in the experimental group were larger than those of the con- trol group.

CONCLUSIONCGFe can efficiently stimulate the proliferation, differentiation and extension of MC3T3-E1 cells.

?Objective: To study the effects of endogenous bone morphogenic protein 2(BMP 2) and 6(BMP 6) in rat facial nerve after injury. Methods: Facial nerve trunk was cut and then anastomosed in 30 adult male Sprague Dawley rats,another 6 rats without operation were used as the controls. Every 6 rats were killed 6 ,72 h,1, 2 and 4 weeks respectively after operation and nerve specimens were immunohistochemically examined for BMP 2 and BMP 6 expression. Result:In the control nerve the gray level of BMP 2 and BMP 6 were 210.89?8.21 and 232.03?9.25 respectively. 6, 72 h,1, 2 and 4 weeks after operation the gray level of BMP 2 were 242.83?11.01,240.67?7.91,234.46?5.28,232.12?7.27 and 220.71 ?10.19;that of BMP 6 210.75?5.19,204.08?10.85,198,91?8.58,186.37?4.11 and 184.62? 8.45 ,respectively. Conclusion: BMP 2 may play a role in the early irritable actions when facial nerve was injured. However, BMP 6 might have functions in the regeneration of facial nerve during later period after injury.

OBJECTIVE:To determine the plasma mycophenolic acid concentration by HPLC, and study the multidoses pharmacokinetics character of mycophenolic acid in Chinese kidney transplantation patients. METHODS: The samples were precipitated with acetonitrile before injection. Diamonsil C18 column was used. The mobile phase consisted of acetonitrile-10 mmol?L-1 KH2PO4 (5∶6) at a flow rate of 1.1 mL?min-1. The detection wavelength was 254 nm and the column temperature was 40 ℃. This method was used to determine the multidoses pharmacokinetics in 12 kidney transplantation patients. RESULTS: MPA was well-separated from internal standard in chromatography, and endogenous foreign substance in plasma had no interference on the determination. The liner range for MPA was 0.38～59.00 ?g?mL-1,and the lowest detectable concentration of MPA was 0.38 ?g?mL-1. The recovery rate stood at 89.32%～97.63%; Both intra-day and inter-day RSD were less than 8.00%. Significant individual difference was noted among the patients treated with MMF in pharmacokinetic results, which was in line with the literature. CONCLUSION: This method is accurate and simple and applicable for the pharmacokinetics study of mycophenolic acid.

Objective To evaluate the antidepressant efficacy and adverse reactions of 3 different stimulus dosage of modified electroconvulsive therapy ( MECT) in patients with depressive episode. Methods 120 patients with depressive episode were randomized into low dosage group ( n=40) ,medium dosage group ( n=40) and high dosage group ( n=40) . Low dosage were 50%× age,medium dosage was 70%×age,high dosage was 80%×age . All patients received 6 treatments. Hamilton depression scale?17(HAMD?17) was used to evaluate the clinical symptoms at baseline,3 and 6 treatments. Effects and adverse reactions were compared among three groups.Results Compared with baseline the HAMD?17 scores of the 3 groups were significantly improved after 6 treatments( t=24.026, P=0.000;t=26.541, P=0.000;t=25.904, P=0.000) , but there were no statistically significant differences among the three dosage groups(F=0.409, P=0.665). Compared with low dosage group((27.2±5.4)%),the HAMD?17 scores reductive ratio of medium dosage group((34.3±6.8)%) and high dosage group((33.9±6.9)%)) were significantly improved after 3 treat?ments ( t=-5.513, P=0.000;t=-4.785, P=0.000). Compared with the low dosage group,the incidence rate of headache,nausea and vomiting,delirium were significantly higher in high dosage groups( headache:χ2=14.532, P=0.000;nausea and vomiting:χ2=13.333, P=0.000;delirium:χ2=14.907, P=0.000) . The re?covery time was significantly longer in medium dosage group ( ( 10. 5 ± 1. 6 ) min ) and high dosage group ((11.2±1.8)min) than that in low dosage group((8.8±1.2)min)( t=-5.144,=0.000;t=-6.889, P=0.000).Conclusion Different stimulus dosage of MECT for depressive episode has equivalent curative effect after 6 treatment . Medium dosage and high dosage treatments appear to have an early onset of efficacy,but may also be associated with more adverse reactions.

Objective To explore the expression of small GTP-binding protein TC10 during nerve injury and construct eukaryotic expression vector carrying rat TC10 cDNA. Methods Total RNA was extracted from rat facial nucleus, and the change of rat TC10 full-length cDNA was investigated by RT-PCR. PCR product was inserted into eukaryotic expression vector pcDNA3 to construct the pcDNA 3-TC10. The plasmid was identified by restriction enzyme analysis and sequencing analysis. Results After injury, the expression of TC10 rapidly increased, Restriction enzyme assay and sequencing assay showed that the construct we got was rat TC10 full-length cDNA construct.

Objective:In order to investigate the role of microRNA in the pathogenesis of ankylosing spondylitis patients,we detected the peripheral blood of patients with ankylosing spondylitis in miR-17,miR-181,miR-106,miR-30 and miR-495 expression, thereby to study the role of microRNA regulation of clinical and diagnostic value in the pathogenesis of ankylosing spondylitis provide new ideas.Methods:Collection of patients with ankylosing spondylitis and normal peripheral blood,peripheral blood mononuclear cells were isolated and extracted PBMC small RNA,using primers specific stem-loop reverse transcribed into cDNA,and build a mature miR-17,miR-181,miR-106,miR-30 and miR-495 T-carrier,standard curve,the use of stem-loop method by Real-time PCR technology to detect miR-17,miR-181,miR-106,miR-30 and miR-495 expression level.Results: In this study,92 cases of clinical samples were collected,of which 61 patients with AS,normal 31 cases.By Real-time PCR detection showed that compared with normal subjects,there was upregulation of miR-106 (P>0.05) and miR-30 (P>0.05) in the peripheral blood of patients with ankylosing spondylitis;down-regulation were miR-181 ( P<0.05 ) , miR-495 ( P<0.05 ) and miR-17 ( P>0.05 ) , in which the miR-181 and miR-495 was statistically significant.Conclusion:Compared with normal, there are differences in the peripheral blood of patients with ankylosing spondylitis expression of miR-495 and miR-181,which may be targeted to regulate TLR-4,HLA-B,DVL,GSK/3βgenes,this may be found in the pathogenesis of ankylosing spondylitis studies provide new ideas,to become a new clinical diagnosis markers.

Objective To investigate the relations between adiponectin(APN) and lipoprotein associated phospholipase A2 (LP‐PLA2) with the development and progress of essential hypertension .Methods 60 patients with essential hypertension were collect‐ed and divided into 3 groups of the hypertension grade 1 ,2 ,3 groups according to the levels of blood pressure ,20 cases in each group .Contemporaneous 20 healthy controls were selected .The peripheral venous blood samples were collected from the cubital vein and the serum levels of APN and LP‐PLA2 were measured by ELISE .The related biochemical indicators were simultaneously detected .Results The serum APN levels in the hypertension grade 1 ,2 ,3 groups were significantly lower than that in the control group ,moreover which in the hypertension grade 3 group was significantly lower than that in the hypertension grade 1 group(P<0 .05);on the contrary ,serum levels of LP‐PLA2 in the hypertension grade 1 ,2 ,3 groups were significantly higher than that in the healthy control group ,moreover which in the hypertension grade 3 group was obviously higher than that in the hypertension grade 1 group(P<0 .05);serum APN was significantly negatively correlated with LP‐PLA2(r= -0 .772 ,P<0 .05) .Conclusion Serum levels of APN and LP‐PLA2 are closely related with the development and progress of essential hypertension .

Objective: To investigate the effect of matrine (Ma) on telomerase activity and cell cycle in hepatoma cell line HepG 2 cells. Methods: TRAP ELISA method was used to determine the telomerase activity in HepG 2 cells which were treated with different concentrations of Ma. Plasmid inserted with 800 bp of the human telomerase reverse transcriptase (hTERT) promoter was transiently transfected into HepG 2 cells by lipofect. Different concentrations of Ma were added into culture media 2 h later, and the activity of the hTERT promoter was tested 48 h after transfection. In addition, the percentages of HepG 2 cells in different cell cycle were determined by the flow cytometry on the 24, 48 and 72 h respectively after adding the different concentrations of Ma. Results: The telomerase activity of HepG 2 was suppressed by Ma at the dose of 750 ?g/ml and the expression of hTERT promoter was also inhibited. The percentage of G 0/G 1 stage cells increased and the percentage of S and G 2/M stages cells decreased in both 500 ?g /ml and 750 ?g /ml groups 48 and 72 h after Ma was added. Conclusion: Ma may have inhibitory effect on hTERT promoter expression, which is related to the telomerase activity and cell cycle regulation.

The purpose of this study is to design push-pull osmotic pump (PPOP) tablets of famotidine using the expert system for the formulation design of osmotic pump of poor water-soluble drug which had been established by the authors. Firstly, the parameters which were requisite of the system input were obtained from literatures and experimental tests. Then the parameters were input into the system, and the program was run. The system displayed the designed formulations sequential. Finally, famotidine PPOP was prepared according to the designed formulations and the in vitro dissolution was carried out. It was found out that the target formulation of famotidine PPOP which could release for 24 hours was obtained in a very short period. Meanwhile, the practicability of the established expert system was proved.