Objective To construct eukaryctie expression vectors stably expressing Smad-7 and study its function as TGF-β1 inhibi- tor. Methods Smad-7 genes were obtained by PCR from fetal liver cDNA library and inserted in to eukaryotic expression plasmid pcD- NA3. 1/myc-his-B (-). The constructed plasmids containing right sequence of target genes were transfected into human mesangial cells by lipofectamine. The ceils were selected by G418, the expression of Smad-7 was detected by Western-blotting. Cell proliferation was detected by MTT method, and cell cycle was detected by Flow Cytometer to study its function as TGF-β1 inhibitor. Results We successfully con- struct Smad-7 expression plesmid. Smad-7 can inhibit the effects of TGF-[M induced GI cell cycle block and apoptosis. Conclusion The eukaryotic expression vector of Smad-7 has been successfully constructed. Over-expression of exogenous Smad-7 can inhibit the effects of TGF-β1 induced GI cell cycle block and apoptosis.

Peripheral blood mononuclear cells from 32 cases of osteosarcoma and 20 normal controls were separated and induced by lipopolysaccharide, followed by 48 hour incubation in vitro, then the supernatant were collected. The levels of IL-6 were measured by enzyme linked immunosorbent assays, the concentrations of NO were measured by Griess methods. The results were as follows: The concentrations of IL-6 and NO were significantly higher than those in normal contrls (P<0.01). There were positive correlation between the levels of IL-6 and NO in patients with osteosarcoma (r=0.652, P<0.01). The results suggest that the immune function of peripheral mononuclear cell in patients with osteosarcoma was disordered. It may activate peripheral mononuclear cells to produce high levels of IL-6 and NO, which may take part in the pathogenesis of osteosarcoma.

Clinical laboratory biological safety is one of whole society safety. This paper introduced briefly the current situation of clinical medical laboratory biosafty in the hospital. and set forth common biological hazards specifically for whose characteristics. Combining the biosafety administration measures from abroad, the issue of laboratory biological safety administration was considered, and put forward some suggestions according to related law and regulation of national laboratory safety administration in order to strengthen clinical laboratory biosafety administration.

OBJECTIVE: To explore the effect of hepatocyte growth factor (HGF) on inducible nitric oxide synthase (iNOS), NO and interleukin-1β (IL-1β) in the cerebrum of rats subjected to cerebral ischemia/reperfusion (I/R). METHODS: Sprague-Dawley rats were randomly divided into 5 groups: a sham group, an I/R group,an HGF1 group, an HGF2 group, and an HGF3 group. The latter 3 groups were respectively injected 15, 30 and 60 μg/kg HGF. The focal cerebral I/R model was established by sutureoccluded method. After 1.5 h ischemia followed by 24 h reperfusion, the iNOS activity and NO content in the ischemic cerebral tissue were assessed. The expression of iNOS mRNA and IL-1β mRNA was detected. The level of iNOS protein and IL-1β content were determined. In addition, cultured cerebral cortical neurons in vitro were exposed to I/R. Then the expression of iNOS and IL-1β protein in the neurons was detected, and NO content was assessed. RESULTS: The iNOS activity and NO content in the ischemic cerebral tissue were increased. The expression of iNOS mRNA and IL-1β mRNA was upregulated. The level of iNOS protein and IL- 1β content were increased. Administration of HGF decreased the iNOS activity and NO content, and downregulated the expression of iNOS mRNA, IL-1β mRNA, iNOS protein and IL-1β content in the ischemic cerebral tissue. HGF decreased the expression of IL-1β, iNOS protein and NO content in the cortical neurons exposed to I/R in vitro. CONCLUSION HGF can inhibit the expression of IL-1β and decrease the expression of iNOS and content of NO, which is probably one of the mechanisms mediating the protection of HGF against cerebral ischemia injury.
Animals ; Brain Ischemia ; metabolism ; Cerebrum ; metabolism ; pathology ; Down-Regulation ; Hepatocyte Growth Factor ; pharmacology ; Interleukin-1beta ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; Up-Regulation

Objective To observe the protective effect of electroacupuncture(EA)on ankle joint synovitis and HIF-1 α/MDM2/p53 signaling pathway in adjuvant arthritis(AA)rats.Methods SD rats were randomly divided into control group,model group,sham EA group,agonist group,EA group,EA+antagonist group,antagonist group,with 10 rats in each group.Preparation of AA rat model by injecting Freund's complete adjuvant into bilateral foot pads of rats.EA(2 Hz,3 V)was applied to bilateral ST36 and GB39 or two control points(5 mm left to ST36 and GB39)for 15 min,once every other day for a total of 8 times.HIF-1 α agonists and antagonists were injected into the tail vein of rats in the corresponding groups.The inflammatory conditions of the ankle joint were observed by HE staining,and the contents of serum HIF-1 α was assayed by ELISA.mRNA expression of HIF-1α,VEGF,MDM2 and p53 were detected by RT-PCR,protein expression of HIF-1α,VEGF,MDM2 and p53 were detected by Western blot in synovium of AA rats.Results The results of HE staining showed that compared with the control group,the synovial cells in the model group had significant proliferation and inflammatory cell infiltration(P<0.01);Compared with the model group,the arthritic reaction of the rats in the EA group,the EA+antagonist group and the antagonist group was significantly reduced(P<0.01),while the arthritic reaction of the rats in the sham EA group and the agonist group was still serious(P>0.05).The results of ELISA showed that compared with the control group,the serum HIF-1αexpression of the model group rats was significantly increased(P<0.01);Compared with model group,serum HIF-1αexpression of rats in EA group,EA+ antagonist group and antagonist group was significantly decreased(P<0.01);There was no significant difference in serum HIF-1αexpression among the model group,sham EA group and agonist group(P>0.05).RT-PCR results showed that compared with the control group,the mRNA expression of HIF-1α,VEGF,MDM2 and p53 of model group rats increased significantly;Compared with model group,the mRNA expression of HIF-1α,VEGF and MDM2 of rats in EA group,EA+antagonist group and antagonist group decreased significantly,while the mRNA expression of p53 increased significantly.Western blot results showed that compared with the control group,the protein expression of HIF-1α,VEGF,MDM2 and p53 of model group rats increased significantly;Compared with model group,the mRNA expression of HIF-1α,VEGF and MDM2 of rats in EA group,EA+antagonist group and antagonist group decreased significantly,while the mRNA expression of p53 increased significantly.Conclusion Electroacupuncture intervention has significant protective effect on ankle inflammation in AA rats,HIF-1 α/MDM2/p53 signaling pathway plays a key role in this process.

To explore the effect of epigallocatechin gallate (EGCG) on oxidative stress and Nrf2/HO-1 pathway in neurons subjected to oxygen-glucose deprivation/reperfusion (OGD/R).
 Methods: Primary cultured cerebral cortical neurons were prepared from Sprague-Dawley rats, and the OGD/R cell model was established. After pretreatment with EGCG at different concentrations (12.5, 25.0, 50.0 or 100.0 μmol/L), the neurons were subjected to OGD/R. The cell viability, reactive oxygen species (ROS) level and malondialdehyde (MDA) content were assessed after reperfusion. The superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities were measured. The expression of Nrf2 protein in nucleus, HO-1 mRNA and protein were detected.
 Results: OGD/R treatment reduced the cell viability, elevated ROS level and MDA content, decreased SOD and GSH-Px activities. The expression of Nrf2 protein in nucleus, HO-1 mRNA and protein were increased (P<0.01). Pretreatment with EGCG promoted the survival of neurons exposed to OGD/R, decreased ROS level and MDA content while increased SOD and GSH-Px activities. The levels of Nrf2 protein in nucleus, HO-1 mRNA and protein were upregulated (P<0.01).
 Conclusion: EGCG can reduce the oxidative stress of neurons subjected to OGD/R, which may be related to activation of Nrf2/HO-1 signal pathway and enhancement of the antioxidant ability of neurons.
Animals ; Catechin ; analogs & derivatives ; pharmacology ; Cell Survival ; drug effects ; Cells, Cultured ; Gene Expression Regulation ; drug effects ; Glucose ; Heme Oxygenase-1 ; genetics ; metabolism ; NF-E2-Related Factor 2 ; genetics ; metabolism ; Neurons ; drug effects ; Neuroprotective Agents ; pharmacology ; Oxidative Stress ; drug effects ; Oxygen ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control