Objective: To establish the quatity standard of Sedum aizoon L..Methods: The micro-method and TLC were used for qualitative identification,and a HPLC analysis was applied for quantitative determination of quercetin.Results: A qualitative analysis for Sedum aizoon L.was set up,a good linear relationship was obtained over the range of 1.216-2.16?g/ml and regression equation was Y=40386X-14138(r=0.9991).The average recovery rate was 102.08%(RSD=0.92%).Conclusion: The method is simple,accurate and suitable for quality control of Sedum aizoon L.

BACKGROUND: Changes of elastic fibers in middle cerebral artery(MCA)is close related withthe aged cerebrovascular disease.OBJECTIVE: To observe the changes of elastic fibers of MCA in different aging rats.DESIGN: A descriptive and controlled study based on experimental animals.SETTING: Department of anatomy and central laboratory in a university.MATERIALS: Totally 36 healthy Wistar rats with either gender, weighing 200 - 280 g, were selected from the Animal Institute of the third medical military university of Chongqing[certification SCXX (army) 2002-007].INTERVENTIONS: Changes of elastic fibers of MCA of different aging rats were observed with light microscope, transmission electron microscope and image analysis system. MAIN OUTCOME MEASURES: ①) Major outcome: changes of elastic lamella in MCA of different aging rats; ②) Secondary outcome: ultramicrostructural changes of internal lamella under the transmission electron microscope. RESULTS: With the increase of age, the folded extent and quantity of internal elastic lamella were decreased, and the content of elastic fibers were also decreased significantly( P ＜ 0.01 ). However, the ratio of collagen fibers to elastic fibers was increased significantly( P ＜ 0.01 ) . In the aging group above 24 months, the internal elastic lamina thinned, delaminated and disrupted, and the lipid deposited in it. Endothelial cells and smooth muscle cells passed through the internal elastic lamina. CONCLUSION: Changes of elastic fibers may be related with the increased susceptibility to the cerebrovascular disease in aged people.

OBJECTIVETo explore the effects of hepatitis B virus X gene and p53 on hepatocellular growth.

METHODSTwo kinds of plasmids containing sense and antisense human wild p53 gene respectively were constructed. SMMU-7721 cells were transfected with HBx, sense-wtp53 antisense-wtp53 separately or cotransfected with either HBx and sense-wtp53 or HBx and antisense-wtp53. Flow cytometry was adopted to measure the apoptosis rates and the effects of HBx on cell cycle progression. The activity of p21(Waf1) promoter-luciferase construct was detected. Growth curves for SMMU-7721 stably transfected with pcDNA3 and pcDNA3HBx were analyzed.

RESULTSAfter doxorubicin administration, HBx was noticed able to initiate apoptosis of the liver cells. The apoptosis rate was 5.32% in the pcDNA3 transfected and 12.66% in the pcDNA3HBx transfected groups respectively. HBx could also abrogate p53-mediated apoptosis. The apoptosis rate in groups transfected with pcDNA3, pcDNA3wtp53 and pcDNA3HBx + pcDNA3wtp53 was 5.32%, 11.72% and 4.67% respectively. In compared with the normal group, the number of cells in transiently HBx-expressed group and HBx-transfected group decreased 4.79% and 10.25% respectively. HBx inhibited the activity of p21(Waf1) promoter-luciferase constructed (P < 0.05) and promoted cell growth. The growth rate of HBx expression cells was faster.

CONCLUSIONUnder DNA damage, HBx reduced expression of p21(Waf1) by repressing the activity of p53 protein, followed by disturbing the regulation of G(0)-G(1) cell cycle checkpoint, and promoted the growth rate of hepatoma cells.

Apoptosis ; Carcinoma, Hepatocellular ; pathology ; virology ; Cell Division ; Cell Line, Tumor ; Genes, p53 ; Hepatitis B Antigens ; biosynthesis ; genetics ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; pathology ; virology ; Trans-Activators ; biosynthesis ; genetics ; Transfection ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics

Animals ; Carcinoma, Hepatocellular ; genetics ; metabolism ; virology ; Genes, p53 ; Hepatitis B ; genetics ; metabolism ; virology ; Hepatitis B virus ; Humans ; Inhibitor of Growth Protein 1 ; Intracellular Signaling Peptides and Proteins ; metabolism ; Liver Neoplasms ; genetics ; metabolism ; virology ; Loss of Heterozygosity ; Nuclear Proteins ; metabolism ; Point Mutation ; Signal Transduction ; Trans-Activators ; genetics ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Tumor Suppressor Proteins ; metabolism

Most plants can form a symbiosis in root with microorganisms for mutual benefit, Nonlegumes mainly form the symbiotic mycorrhiza with arbuscular fungi. The interaction is initiated by invasion of arbuscular mycorrhizal (AM) fungi into the plant root, and follows by production of several special signal molecules, such as the symbiosis receptor-like kinase (SYMRK) from plant. SYMRK has an extracellular domain comprising three leucine-rich repeats (LRRs), a transmembrane domain and an cytoplasmic protein kinase domain. Symrk is required for a symbiotic signal transduction pathway from the perception of microbial signal molecules to the rapid symbiosis-related gene activation. Study of symrk may set up a solid foundation for giving further insight on the function and mechanism of plant-fungi symbiosis.
Amino Acid Sequence ; Host-Pathogen Interactions ; Lycopersicon esculentum ; Molecular Sequence Data ; Mycorrhizae ; physiology ; Phosphotransferases ; classification ; genetics ; Phylogeny ; Plant Proteins ; classification ; genetics ; Plant Roots ; enzymology ; genetics ; microbiology ; Sequence Homology, Amino Acid ; Signal Transduction ; genetics ; Symbiosis ; genetics

Cell Line, Tumor

Objective To explore the effects of different anesthesia methods on postoperative TLR4 expression and the level of serum S100β protein in children.Methods Totally 122 children with elective surgery were randomly divided into two groups.The control group was treated with ketamine intravenous anesthesia,while the observation group was treated with inhalation anesthesia with sevoflurane.Expression of TLR4,the level of serum S100β protein and the adverse reactions of anesthesia were observed.Results During the period of anesthesia,the incidence rates of physical and mental symptoms in the observation group were significantly lower than the control group (P ＜ 0.05).The levels of serun S100β protein at 2 h and 24 h after operation in the observation group were significantly lower than those in the control group (P ＜ 0.05).Compared with those before operation,there was no significant difference in the expression of TLR4 at the time of skin incision,2 h and 24 h after operation (P ＞ 0.05).The expressions of TLR4 at the time points of after skin incision,2 h and 24 h after operation in the control group were significantly lower than those before operation and the observation group (P ＜ 0.05).Conclusion The sevoflurane inhalation anesthesia can reduce the adverse reactions of anesthesia in children with surgery.

Objective To explore the effects of different anesthesia methods on postoperative TLR4 expression and the level of serum S100β protein in children.Methods Totally 122 children with elective surgery were randomly divided into two groups.The control group was treated with ketamine intravenous anesthesia,while the observation group was treated with inhalation anesthesia with sevoflurane.Expression of TLR4,the level of serum S100β protein and the adverse reactions of anesthesia were observed.Results During the period of anesthesia,the incidence rates of physical and mental symptoms in the observation group were significantly lower than the control group (P ＜ 0.05).The levels of serun S100β protein at 2 h and 24 h after operation in the observation group were significantly lower than those in the control group (P ＜ 0.05).Compared with those before operation,there was no significant difference in the expression of TLR4 at the time of skin incision,2 h and 24 h after operation (P ＞ 0.05).The expressions of TLR4 at the time points of after skin incision,2 h and 24 h after operation in the control group were significantly lower than those before operation and the observation group (P ＜ 0.05).Conclusion The sevoflurane inhalation anesthesia can reduce the adverse reactions of anesthesia in children with surgery.

OBJECTIVESTo study the interaction of hepatitis virus B (HBV) and tumor suppressor p53.

METHODSPlasmid pCMVp53 was transfected or cotransfected with pCMVHBVa (wild type HBV) or PCMVHBVb (mutant type HBV) into the hepatoma cell line SMMC-7721 by lipofectamine. Apoptosis cells were labeled by annexin V-FITC and confirmed by flow cytometry. Reporter plasmids PG13-CAT or p21-luc were cotransfected respectively in each group to indicate transactivation activity of p53 and it's effect on p21 promoter. Western blot was performed to observe p53 expression in each group.

RESULTSThe group transfected by pCMVp53 alone exhibit higher luciferase activity and higher apoptosis rate, otherwise, p53 expression, enzyme activity of PG13-CAT or p21- luc and cell apoptosis rate were much higher in the group cotransfected by pCMVp53 and pCMVHBVa, but not in the other cotransfected group; HBV replication was enhanced in p53 cotransfected group.

CONCLUSIONp53 expression and effects could be enhanced by HBV and p53 had positive regulation effect on HBV replication.

Apoptosis ; Carcinoma, Hepatocellular ; genetics ; pathology ; virology ; Cell Line, Tumor ; Hepatitis B virus ; genetics ; physiology ; Humans ; Liver Neoplasms ; genetics ; pathology ; virology ; Luciferases ; metabolism ; Promoter Regions, Genetic ; Proto-Oncogene Proteins p21(ras) ; genetics ; Trans-Activators ; genetics ; Transfection ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Virus Replication

OBJECTIVETo explore the effects of hepatitis B virus X protein (HBx) on hepatoma cell growth through p14(ARF)-dependent and p14(ARF)-independent pathways.

METHODSHBx and p14(ARF) were transfected either separately or in combination into HepG2 cells containing wt-p53 but not expressing p14(ARF). The cells were divided into 4 groups, namely pcDNA3 (control), pcDNA3HBx, pcDNA3p14(ARF), and pcDNA3HBx + pcDNA3p14(ARF) groups. Flow cytometry was used to examine the apoptosis rates and cell cycle progression of HepG2 cells in different groups. The expression of p14(ARF), MDM2, p53, and p21(WAF1) proteins were investigated by detecting the activity of p21(WAF1) promoter-luciferase and using Western blotting.

RESULTSThe apoptosis rates of HepG2 cells in pcDNA3HBx and pcDNA3p14(ARF) groups were significantly higher than that in the control group (14.11%, 13.72% vs 10.66%). Compared with the control group, pcDNA3HBx and pcDNA3p14(ARF) groups also showed significantly higher cell percentages arrested at G(0)/G(1) phase (63.62%, 61.75% vs 57.42%), luciferase activity of p21 promoter (1.25-/+0.05, 1.09-/+0.06 vs 0.77-/+0.03) and expressions of p53 and p21(WAF1). The cell apoptosis rate, percentage of cells in G(0)/G(1) phase and expression level of p14(ARF) were even higher in pcDNA3HBx+pcDNA3p14(ARF) group (18.61%, 66.74%, and 3.53-/+0.43, respectively) than in either p14(ARF) or HBx group.

CONCLUSIONHBx induces p53 expression through p14(ARF)-dependent and independent pathways to activate p21(WAF1) promoter, leading to G(0)/G(1) arrest and apoptosis of HepG2 cells.

Carcinoma, Hepatocellular ; genetics ; pathology ; virology ; Cell Line, Tumor ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; Humans ; Liver Neoplasms ; genetics ; pathology ; virology ; Promoter Regions, Genetic ; Trans-Activators ; genetics ; Transfection ; Tumor Suppressor Protein p14ARF ; genetics ; Tumor Suppressor Protein p53 ; genetics