Objective This study was performed to investigate the effect of hypoxia on glucose-6-phosphate isomerase (G6PI) expression and cell cycle of fibroblast-like synoviocytes from synovium of rheumatoid arthritis (RA) and osteoarthritis (OA) under hypoxia or normoxia.Methods Fibroblast-like synoviocytes were cultured with either of hypoxia (3％ oxygen) or normoxia (21％ oxygen) for 24 hours.The mRNA expression of G6PI and HIF-1α was tested by PCR quantification,while the protein levels of G6PI and HIF-1α were measured by western blot.Cell cycle was performed by FACS.T-test and Mann-Whitney U were used for statistical analysis.Results The expression levels of G6PI mRNA under hypoxia in RA were higher than those of OA (2.6±0.4 vs 1.5±0.4,P＜0.05).The protein levels of G6PI in RA were higher than those of OA (P＜0.05).The expression levels of HIF-1α mRNA under hypoxia in RA were higher than those of OA (2.9±0.8vs 1.4 ±0.4,P＜0.05).The protein levels of HIF-1α in RA were higher than those of OA (P＜0.05).The G1 phase ratio of cell cycle was decreased significantly under hypoxia than those of normoxia in RA ELs (t=1 1.31,P＜0.05).The S and G2 phase ratio of cell cycle were increased.Conclusion Hypoxia upregulates G6PI and HIF-1α expression and improves proliferation in fibroblast-like synoviocytes.

Animals ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; Cell Differentiation ; Cell Proliferation ; Humans ; Liver Cirrhosis ; pathology ; surgery ; Liver Function Tests ; Mesenchymal Stem Cell Transplantation ; Rats ; Stem Cells ; cytology

AIM To explore the role of nitric oxide (NO) mediators in seizure behavior and the related expression of interlukin 6(IL 6). METHODS The IL 6 immunoreactivity (IL 6 ir) and behaviour were observed in kainic acid (KA) induced seizure following pretreatment with L nitroarginine( L NNA), a inhibitor of nitric oxide synthase(NOS), or the same number of moles of L arginine( L Arg). RESULTS The results showed that time dependent seizures were induced on animals after administration of KA (10 mg?kg -1 ), accompanied by the immediate enhancement of IL 6 ir in hippocampal structure, piriform area and cortex. The behaviors of rats were not markedly altered by chronic pretreatment with L NNA (50 mg?kg -1 ) or L Arg (40 mg?kg -1 ). However, after the administration of KA, the seizure behaviors were obviously different in the group of L NNA and L Arg respectively. Seizure was agrevated in the animals with L NNA pretreatment and many rats died at KA 3 hours, whereas seizure was alleviated after KA in the group of L Arg. IL 6 expression was apparently up regulated in some brain areas such as hippocampus in the group of KA pretreated with L NNA, but opposite effects appeared in the group pretreated with L Arg before KA injection. CONCLUSION These results indicate that endogenous NO mediators may alleviate the seizure behaviors and may down regulated the early expression of IL 6 in KA challenged seizure, but it the mechanism of the early expression of IL 6 with the homeostasis of endogenous NO mediator merits further study.nismoftheearlyexpressionofIL 6withthehome

Adult ; Aged ; Female ; Hemodynamics ; Hepatitis B, Chronic ; complications ; Humans ; Hypertension, Portal ; diagnostic imaging ; physiopathology ; Liver Cirrhosis ; diagnostic imaging ; physiopathology ; Male ; Middle Aged ; Portal Vein ; diagnostic imaging ; physiopathology ; Ultrasonography, Doppler, Color ; Venous Pressure

AIM To study the chemical constituents from the Polygonum amplexicaule var.sinense.METHODS The n-butanol fraction of ethanol extract from P.amplexicaule var.sinense was isolated and purified by TLC,normal-phase silica,macroporous resin column,sephadex column,preparative TLC and semi-preparative HPLC column,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Ten compounds were identified as 2-(aminomethyl)-4-methoxy-phenol (1),p-methyl-hydroxybenzoate (2),p-methyl-hydroxyphenylacetate (3),2-[2-(methylamino) phenyl]-4-thiazolecarboxylic acid,methyl ester (4),p-methylphenylethanol (5),bergenin (6),arbutin (7),rhododendron-2-O-β-D-glucopyranoside (8),n-butyl gallate (9) and β-sitosterol (10).CONCLUSION Compounds 1-8 are isolated from this plant for the first time.

Adenocarcinoma ; blood supply ; physiopathology ; Capillaries ; metabolism ; ultrastructure ; Cell Separation ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; ultrastructure ; Humans ; Microcirculation ; Stomach Neoplasms ; blood supply ; physiopathology

OBJECTIVETo construct a recombinant adenovirus vector carrying antisense heat shock protein 70 (HSP70) cDNA and observe its effect on inhibiting the growth of laryngeal carcinoma Hep-2 cells.

METHODSThe HSP70 gene fragment encoding the 5' region was cloned reversely into the shuttle plasmid PAdTrack-CMV, and the resultant plasmid was recombined with the backbone plasmid PadEasy-1 in the E.coli Bj5183 cells to generate the recombinant adenovirus vector. The adenovirus were then packaged and amplified in 293 cells, and the viral titer was determined using GFP.

RESULTSThe recombinant adenovirus vector carrying antisense HSP70 cDNA was constructed successfully with a viral titer of 8 x 10(9). HSP70 expression of Hep-2 cells was obviously blocked by antisense HSP70 RNA, and Western blotting and immuohistochemistry demonstrated that cells transfected with antisense HSP70 did not express or express HSP70 at low levels. Flow cytometry presented apoptotic peak in the antisense HSP70-transfected cells, but not in the control cells.

CONCLUSIONThe recombinant adenovirus vector containing antisense HSP70 cDNA can effectively deliver antisense HSP70 gene into Hep-2 cells, suggesting the great potential of this gene therapy strategy in management of human laryngeal carcinoma.

Adenoviridae ; genetics ; Cell Line, Tumor ; DNA, Antisense ; pharmacology ; DNA, Complementary ; genetics ; Genetic Therapy ; Genetic Vectors ; biosynthesis ; HSP70 Heat-Shock Proteins ; genetics ; Humans ; Laryngeal Neoplasms ; therapy ; RNA, Antisense ; pharmacology ; Transfection

OBJECTIVETo investigate the significance of DARPP-32 protein expression in gastric carcinoma tissue and cell lines.

METHODSThe expression of DARPP-32 protein in normal gastric mucosa and gastric carcinoma tissue was evaluated by immunohistochemical staining using streptavidin-biotin complex technique. The expression in gastric carcinoma tissue and cell lines was evaluated by Western blotting.

RESULTSThe expression rate of DARPP-32 protein in gastric adenocarcinoma tissue (92.7%) was significantly higher than that in normal gastric mucosa (52.6%, P < 0.05). There was no significant association between DARPP-32 protein expression and degree of tumor differentiation, local invasion and distant metastasis. As compared with adjacent non-carcinomatous gastric mucosa, both DARPP-32 and its truncated isoform t-DARPP were overexpressed in gastric adenocarcinoma tissue (t = 2.45, P = 0.015); and t-DARPP overexpression was more frequently seen. Expression of DARPP-32 and t-DARPP could also be detected in human gastric cancer cell lines. The expression of DARPP-32 protein was obviously reduced in SGC7901 drug-resistant cell strains.

CONCLUSIONSDARPP-32 is overexpressed in gastric carcinoma. It may play an important role in gastric carcinogenesis. The underlying signal pathways in neoplastic gastric epithelium may also be related to the multi-drug resistance property of gastric cancer cells.

Adenocarcinoma ; metabolism ; Aged ; Antibiotics, Antineoplastic ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Dopamine and cAMP-Regulated Phosphoprotein 32 ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Female ; Gastric Mucosa ; metabolism ; Humans ; Male ; Middle Aged ; Nerve Tissue Proteins ; metabolism ; Phosphoproteins ; metabolism ; Stomach Neoplasms ; metabolism ; Vincristine ; pharmacology

OBJECTIVETo investigate the role of human angiopoietin-1 (Ang1) in tumorigenesis and angiogenesis of human gastric cancer cell line SGC7901 in nude mice.

METHODSRecombinant human Ang1 sense or antisense eukaryotic expression vectors were constructed, and transfected by lipofectin into human gastric cancer line SGC7901. Stable transfectants were obtained respectively, namely 7Ang1+ for sense, 7Ang1- for antisense, and 7901P for empty vector transfected cells. Semiquantitative PCR and Western blot were employed to testify the transfection efficiency. Cell growth curve and cell cycle were observed by MTT assays or flow cytometry. In in vivo study, growth of SGC7901 xeno-transplant was observed in BALB/c nude mice. Microvessel density (MVD) was analyzed by immunohistochemistry for Factor VIII staining.

RESULTSStably transfected cell lines were established and decreased expression of Ang1 protein and mRNA in the antisense transfected SGC7901 cells was achieved. Tumorigenesis of 7Ang1- cells on day 30 days was significantly inhibited with decreased MVD as compared to that in 7901P and 7Ang1+ cells (P < 0.01).

CONCLUSIONAngiopoietin-1 plays an important role in tumorigenesis and angiogenesis of gastric cancer which can be partially abrogated by antisense technique.

Angiopoietin-1 ; biosynthesis ; genetics ; Animals ; Cell Line, Tumor ; DNA, Antisense ; genetics ; Genetic Vectors ; Humans ; Mice ; Mice, Nude ; Microcirculation ; pathology ; Neoplasm Transplantation ; Neovascularization, Pathologic ; RNA, Messenger ; biosynthesis ; genetics ; Stomach Neoplasms ; blood supply ; metabolism ; pathology ; Transfection

OBJECTIVETo observe blood lead level and related risk factors among children aged 0-6 years old living in cities in China and to provide data for policy development to the prevention on environmental lead pollution.

METHODSA stratified-clustered-random sampling method was used. 17 141 peripheral blood samples of 0-6 years old children from 15 cities in China were tested. Tungsten atomizer absorption spectrophotometer was employed to determined the blood lead level. Related factors were also studied using a standardized questionnaire. Data were analyzed on related risk factors that affecting blood lead levels through multiple regression method.

RESULTSThe mean blood lead level of 0-6 years old children from 15 cities in China was 59.52 microg/L including 10.45% of those > or =100 microg/L, and 0.62% > or = 200 microg/L. However, the blood lead levels seemed to have had a trend of increase parallel to age among 0-6 years-old children and were higher for boys (59.50 microg/L) than girls (54.95 microg/L). The risk factors which influenceing children's blood lead levels would include the type of housing, parent's education levels, social status and hobby, children's behavior habit, dieting habit and nutritional condition.

CONCLUSIONThe blood lead levels of children in China were lower than data gathered from former national studies but higher than those from developed countries, suggesting that the. Government and the whole society should be aware of the problem on lead poisoning among children during their childhood.

Child ; Child, Preschool ; China ; Female ; Health Policy ; Humans ; Infant ; Infant, Newborn ; Lead ; blood ; Logistic Models ; Male ; Quality Control ; Risk Factors ; Sampling Studies ; Urban Population ; statistics & numerical data