Objective To investigate the effect of methyltransferase inhibitor 5-Aza-dC (5-aza-2’-deoxycytidine) on proliferation and activation of myofibroblasts in arthrofibrosis of the knee. Methods The model of arthrofibrosis of rat knee was firstly established.Myofibroblasts were isolated and cultured from the posterior capsule of rat knee. Cells were identified by immunofluorescence.Myofibroblasts were treated with 2 μmol/L 5-Aza-dC. The expression levels ofα-SMA、col1A1 mRNA and protein in myofibroblasts before and after the treatment with 5-Aza-dC were detected by RT-PCR and Western blotting,respectively.The proliferation rate of these myofibroblasts were detected by MTT method,and the cell cycle was detected by flow cytometry of DNA content in each phase. Results The extension of rat knee in arthrofibrosis model was significantly decreased. α-SMA protein,the representational protein of myofibroblast,was largely expressed in the isolated cells from model knees. The expression levels ofα-SMA、col1A1 mRNA and protein in myofibroblasts were decreased after the treatment with 5-Aza-dC and the growth of myofibroblasts was also slowed down. Conclusion The methyltransferase inhibitor 5-Aza-dC may become a potential therapeutic drug in the treatment of arthrofibrosis of the knee through inhibiting the proliferation of myofibroblasts.

Toxoplasma gondii rhoptry proteins (ROPs) include dozens of proteins secreted by tachyzoites ,these ROPs are known to be critical for several processes such as invasion ,parasitophorous vacuole establishment ,proliferation ,egress and getting out of host immune protection .ROPs belong to the protein kinase family and have classic kinase domain ,however ,most of the members are designated as pseudokinases due to the lack of critical catalytic residues that are normally required for phos-phorylation of host signal molecules .Recent studies indicated that pseudokinases also have important functions ,such as regula-ting kinase activity by allosteric and promoting the interactions between proteins as scaffolds .In this paper ,we summarize the pseudokinases structural characteristics and some pseudokinases functions ,for the study of toxop lasma gondii and host interac-tion mechanisms ,molecular vaccines screening and drugs design .

Objective To explore the application of experienced teaching method in the teaching of Fundamentals of Nursing.Methods The feasibility and validity of experienced teaching method was evaluated by expounding the utilization and result of role-play method and practice experienced method.Results The experienced teaching method received good result and proved appropriate in the teaching of Fundamentals of Nursing.Conclusion The experienced teaching method overcame the disadvantages of mere traditional teaching method and aimed at the improvement of quality and ability of students.It contributed to developing the critical thinking ability and other abilities of students.

[Objective]To observe whether erythromycin and alendronate are effective in treating aseptic loosening in a murine osteolysis model. [Method]Air pouches were established in the back of BALB/c mice by injecting sterile air subcutaneously. After the air pouches were mature, grafts of calvaria from syngeneic mouse donors were implanted in the air pouches and polyethylene debris was injected into the air pouches. Mice were divided into 4 groups. In the first group, saline was injected into the air pouches.In the other three groups, polyethylene debris was used. Saline,erythromycin and alendronate were injected intraperitoneally separately.Pouch tissues were collected at 14 days after polyethylene debris administration for molecular and histologic analysis.[Result] Both erythromycin and alendronate inhibited osteoclastogenesis and bone resorption, but alendronate was not as effective as erythromycin in anti-inflammation.[Conclusion]Both erythromycin and alendronate can be promising drugs for the treatment of aseptic loosening.

Objective To investigate the transfection and expression of pcDNA3 1-VEGF 165 plasmid in mammalian cells. Methods pcDNA3 1-VEGF 165 plasmid was transfected into CHO cells by electroporation and the positive clones were screened by G418. The expression of VEGF 165 gene in the transfected CHO cells were detected by RT-PCR, ELISA and western blotting analysis. The biologic activities of VEGF 165 secreted by the transfected CHO cells were measured by Mile's assay and MTT method. Results G418-resistant CHO cell clones were obtained. RT-PCR, ELISA and western blotting analysis showed that there were the transcription and translation of VEGF 165 gene in anti-G418 CHO cells. The mile's assay showed that the expressed VEGF 165 could increase the vessel permeability, and MTT assay proved that it could promote the proliferation of endothelial cells. Conclusion pcDNA3 1-VEGF 165 plasmid could express VEGF 165 with biological activities in mammalian cells.

Objective To establish a animal model for diffusion mesangioproliferation complicating focal segmentally glomerulosclerosis.Methods The models were established in the seventieth day by repetitive injection of 4mg/kg and 3mg/kg adriamycin to the SD rats in the 7th and 30th day after removal of their right kidneys.Results Serious edema, profuse albuminuria, hypoalbuminemia, hyperlipemia and renal failure were presented. Simultaneously, diffuse medium- or serious-grade proliferation of mesangial matrix, slight-grade proliferation of mesangial cells, segmental sclerosis of 22% of glomeruli, and overexpression of PDGF-B and bFGF in glomeruli were showed by pathologic examination, computerized image analysis and immunohistochemistry staining.Conclusions The models established in the transition stage from mesangioproliferation to glomerulosclerosis are helpful to study mechanisms and take measures for prevention and cure of chronic glomerulonephritis.

BACKGROUND:It is difficult to make an exact conclusion about the correlation between atherosclerosis and hemodynamic wal shear stress because of the complexity of the hemodynamic environment in the field of atherosclerosis research.
OBJECTIVE:To analyze the relationship between characteristics of mural coronary artery hemodynamics parameters and atherosclerosis through myocardial bridge-coronary artery analog device.
METHODS:Myocardial bridge-coronary artery analog device was used to simulate experiments in vitro. In the condition of keeping any related parameters unchanged, such as system temperature, the average flow rate and heart rate, we observed and recorded the changes of proximal and distal mean value and oscil atory value of shear stress of mural coronary artery by regulating the oppression level of myocardial bridge to mural coronary artery.
RESULTS AND CONCLUSION:Under 0%oppression, there was no significant difference in distal and proximal mean value and oscil atory value of shear stress of mural coronary artery. Under 50%oppression, the proximal oscil atory value of shear stress was significantly larger than that in the distal end. Under 80%oppression, mean shear stress in the distal end was larger than that in the proximal end. Oscil atory value of shear stress in the proximal end was larger than that in the distal end. With increased oppression, the mean shear stress in the distal end was higher than that in the proximal end, but oscil atory value in the proximal end was higher than that in the distal end. These results indicated that the increase of the oscil atory value of the proximal shear stress of mural coronary artery is an important factor for the occurrence of atherosclerosis.

The Hedgehog (Hh) signaling pathway plays a key role in the embryonic development, which is formed by the Hedgehog(Hh) , membrane-receptor complex Patched (Ptch) and Smoothened (Smo), multifunctional transcription factors Glioma-associated oncogene homolog (Gli) and other signal molecules. The Hh signaling pathway controls a variety of developing processes, however, no or less expression is observed in the normal pancreas tissues.The mutational activation of the Hh signaling pathway is associated with tumorigenesis and metastasis of the pancreatic cancer. The aim of this review is to present a brief overview of the Hedgehog signaling pathway in the mechanism of the tumorigenesis of the pancreatic cancer.

OBJECTIVE:To develop a method for concentration determination of voriconazole in human plasma and apply it in the clinic. METHODS:UPLC-MS/MS method was adopted. Using ketoconazole as internal standard,the determination was per-formed on Shim-pack VP-ODS column with mobile phase consisted of water(containing 1‰ formic acid and 2 mmol/L ammonium acetate)-acetonitrile(gradient elution)at flow rate of 0.3 ml/min and column temperature of 40℃.The electrospray ion source,pos-itive ionizing pattern and multiple reaction monitoring were used;the mass transition ion-pairs of voriconazole and internal standard were m/z 351.2→282.2 and m/z 532.1→490.2. RESULTS:The linear range of voriconazole were 1-10 000 ng/ml (r=0.999 5,n=5),and the limit of quantitation was 1 ng/ml;RSDs of inter-day and intra-day were all lower than 10%;method recovery was higher than 90%(RSD<8%),and extraction recovery was higher than 70%(RSD<8%). The plasma concentrations of voriconazole in 10 patients with invasive fungal infection determined by this method were 507.33-7 011.24 ng/ml,and those of 3 patients were outside the recommended treatment concentration range. CONCLUSIONS:The established method is fast,accurate and sensitive,and can be applied for the therapeutic drug monitoring of voriconazole.

ObjectiveTo explore the effects of fluoxetine on special learning and memory and serum S100B level in depressed model rats.MethodsAdult male SD rats were divided into six groups randomly according random digits table:control group ( A ),depressed model group ( B ),group of depressed model treated with single dose of fluoxetine for one day ( C ),group of depressed model treated with fluoxetine for one week (D),group of depressed model treated with fluoxetine for two weeks (E) and group of depressed model treated with fluoxetine for four weeks (F),ten rats in each group.Except control group,others were subjected to forced-swimming for four weeks,15 min a day.Fluoxetine (10 mg/kg) was given intragastric administration to group C-F before swimming everyday.Morris water maze ( MWM ) was used to measure the spatial learning and memory of rats.ELISA was used to determine the level of serum S100B.ResultsIn the hiding platform test of MWM,there was significant longer of escape latency (EL) in B group than that in A group(P ＜ 0.05 ).And the EL in all groups treated with fluoxetine became shorter with the prolonging of treatment.In the probe test,there were significant longer time in target quadrant in D,E,F than in other quadrant (F =5.162,P ＜ 0.01 ).The levels of serum S100B were lower in E,F groups ( E group ( 0.91 ± 0.23 ) ng/ml,F group ( 0.85 ± 0.21 ) ng/ml) than that in B group (( 1.26 ±0.61 )ng/ml,P＜0.05).ConclusionChronic administration of fluoxetine could improve the impairment of spatial learning and memory and reverse the increase of S100B level in serum of depressed model rats.