Objective To explore the effects and mechanisms of urokinase-type plasminogen activator (uPA) on high glucose-induced rat mesangial cells proliferation and phenotype transformation. Methods Rat mesangial cells were cultured and incubated in media containing either 5 mmol/L D-glucose or 30 mmol/L D-glucose with or without addition of wortmannin, or uPA (105 U/L) for different time periods. At the end of the incubation period, mesangial cells proliferation was assessed by MTT assay and flow cytometric analysis. Cyclin-dependent kinase 2 (CDK2) and p27kip1 expression and activation of Akt were evaluated by Western blotting and Akt kinase assay respectively. Furthermore, the expression and distribution of α-SMA were detected with laser confocal microscopy. Results MTT assay and flow cytometric analysis demonstrated that high glucose induced mesangial cells proliferation (P＜0.05) and an incresed proportion of cells in G2/M+S stage after 24 h incubation (P＜0.01), which were attenuated by uPA or wortmannin (P＜0.01). High glucose induced the enhance of Akt activity after 3 h (P＜0.05), and the effect was inhibited by wortmannin or uPA (P＜0.01). High glucose did not alter CDK2 expression (P＞0.05),but significantly inhibited p27kip1 expression (P＜0.05), which was attenuated by wortmannin or uPA (P＜0.01). High glucose induced the up-regulation of α-SMA expression and perinucleus location in mesangial cells after 24 h (P＜0.01), which were alleviated by wortmannin or uPA (P＜0.01). Conclusion uPA up-regulates p27kip1 expression and counteracts high glucose-induced mesangial cells proliferation and phenotype transformation via blocking PI3K-Akt signaling pathway.

Using gene fusion technology, polypeptide fusion tags can be engineered into target proteins at the genetic level.The resultant recombinant proteins may possess the biochemical properties of the imported fusion tags.Therefore, it is possible to take advantage of fusion tags to improve and evaluate protein expression, to detect and track protein targets, and to purify and characterize proteins.However, it is necessary to eliminate any influence of the fusion tag in structural characterization experiments or in isolating pharmaceutical proteins.Scientists must therefore remove fusion tags prior to structural and functional analyses when fusion tags are suspected of interfering with the biological activity of a protein or influencing its behavior.The fusion tag can be removed by several methods including harsh chemical treatment, mild enzymatic cleavage by endoprotease or exoprotease, and intein-mediated self-cleavage.Here the literature is reviewed in relation to principles, applications, and approaches of each method.

Biopsies were randomly taken from 160 patients through endoscopy. Among them 46 patients suffered from chronic superfacial gastritis, 39 patients from chronic atrophic gastritis, 39 patients from gastric ulcer, and 36 patients from gastric cancer. Ras, p16, and p53 genes were analysed with polymerase chain reaction single strand conformation polymorphism (PCR SSCP). Helicobacter pylori (HP) was examined with RUT. Result: ① the positive rates of ras and mutant p53 in gastric cancer were significantly higher than that in gastritis and gastric ulcer( P

Objective To screen the HBV SPⅡ promoter DNA-binding protein, and to investigate its potential role in the regulation of replication and expression of HBV DNA. Methods By using HBV SPⅡ biotinylated promoter DNA as a selective molecule, the T7 select human liver cDNA library was biopanned and the positive clones were selected. After screening, amplification of positive plaques was performed for inserted DNA fragment and then they were cloned into the pGEM-Teasy vector. Results Four positive plaques were chosen for DNA sequencing. The binding protein of HBV SPⅡ promoter was demonstrated as nicotinamide adenine dinucleotide (NADH) dehydrogenase 4 by BLAST. Conclusion The result suggests that this approach may provide a new tool for the study of replication and expression mechanism of HBV DNA.

Objective Compared with different decellularization procedures for their potential of cell removal and the ability to preserve the matrix. Methods Specimens of bovine pericardiums were treated by 3 approaches (detergent and enzyme extraction、trypsin 、Triton-X 100 and sodium-deoxycholate). Tissue samples were then observed by HE staining and scanning electron microscopy to confirm the removal of cells. Von Gieson(VG) staining and Gomori staining were used for showing the integrity of collagen and elastin. DNA content was examined by the method of DNA extraction. Tissue shrinkage temperature and mechanical properties were also studied. Results Completely decellularization were achieved in 3 groups. While trypsin、Triton-X 100 and sodium-deoxycholate caused severe structural destruction and declined of mechanical properties of the matrix. In contrast, detergent and enzyme extraction achieved completely deeellularization and effectively preservation the matrix structure. Conclusion This research demonstrated detergent and enzyme extraction could achieve both complete decellularization and preservation of the matrix structure. This approach may provide an ideal platform for the construction of tissue-engineering heart valves.

In order to favor the growth and development of the fetus,the maternal-placental-fetal unit is characterized by profound physiologic changes.These adaptations may affect maternal and fetal absorption,distribution,placental transfer,metabolism and excretion of drugs.When evaluating drug use in pregnancy,understanding both pregnancy physiology and the gestation-specific pharmacokinetics of different drugs is necessary to achieve effective treatment and limit maternal and fetal risk.This topic will focus on the pharmacokinetics and safety of drugs during pregnancy in light of changes in maternal physiology and function of placental.

ZJ-XD-I multifunctional internal sterilization machine for medical instrumentation is applied in medical instrumentations with circulatory loop such as anesthetic machine and respirator. The machine can generate ozone and hydrogen peroxide, and then the two gases are mixed and introduced into the internal loop to eliminate adnexed hydrogen peroxide such as the virus and germ, and thus the nosocomial infection due to repeated uses of medical instrumentations. The machine is composed of the gas compression component, atomization unit, gas extraction component, control circuit and the filtrating unit.

The concentration of 5 nucleosides, uracil, uridine, guanidine, adenine and adenosine in culture of Paecilomyces hepialid was determined by the developed method of HPLC. The HPLC method was performed on a Waters SunFire C18 (4.6 mm x 250 mm, 5 μm) column with methanol-water gradient elution as the mobile phase. The detection wavelength was 260 nm and the colunmn temperature was controlled at 30 °C. The linear range was 10.00-200.00 mg · L(-1) (r = 0.9994) for uracil, 10.10-202.00 mg · L(-1) (r = 0.9992) for uridine, 10.00-200.00 mg · L(-1) (r = 0.9991) for guanidine, 10.30-206.00 mg · L(-1) (r = 0.9992) for adenine and 10.45-209.00 mg · L(-1) (r = 0.9991) for adenosine, respectively. The RSD of precision was 0.032%, 0.035%, 0.039%, 0.049%, 0.00080%, respectively. The average recoveries of uracil, guanidine, adenine, and adenosine were 97.34%, 99.10%, 101.6%, 98.61% and 100.2% with RSD of 1.3%, 2.1%, 0.96%, 0.95%, and 1.3% respectively. The method showed high sensitivity, good selectivity, linearity and repeatability, which was suitable for the content analysis of 5 nucleosides components in P. hepialid and its extracts.
Chromatography, High Pressure Liquid ; methods ; Nucleosides ; analysis ; Paecilomyces ; chemistry

Adenoviridae ; genetics ; Cell Death ; Genetic Vectors ; HT29 Cells ; Humans ; Interleukins ; biosynthesis ; genetics ; RNA, Catalytic ; biosynthesis ; genetics ; RNA, Messenger ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection ; Vascular Endothelial Growth Factor A ; metabolism

Animals ; Cell Proliferation ; Cells, Cultured ; Coxsackievirus Infections ; metabolism ; Enterovirus B, Human ; Myocytes, Cardiac ; cytology ; metabolism ; virology ; Osteopontin ; metabolism ; Rats