Objective To compare the effects of isoflurane or sevoflurane in combination with remifentanil anesthesia on blood amyloid beta protein (Aβ) in the elderly patients undergoing abdominal surgery.Methods Two hundred patients of both sexes,aged 65-75 yr,weighing 51-76 kg,of ASA physical status Ⅰ or Ⅱ,scheduled for elective abdominal surgery under general anesthesia,were randomly divided into 2 groups (n =100 each) using a random number table:isoflurane combined with remifentanil anesthesia group (IR group) and sevoflurane combined with remifentanil anesthesia group (SR group).Fifty healthy elderly subjects served as control group (group C).After anesthesia was induced with iv penehyclidine,sufentanil,propofol and vecuronium,the patients were endotracheally intubated and mechanically ventilated.In group IR,anesthesia was maintained with inhalation of isoflurane (end-tidal concentration 1.68 ％,in IR group) or sevoflurane (end-tidal concentration 1.71％,in SR group),and target-controlled infusion of remifentanil (target plasma concentration 2-6 ng/ml).At l day before surgery and 3 days after surgery,the patients' cognitive function was assessed using Mini-Mental State Examination (MMSE),the development of postoperative cognitive dysfunction (POCD) was recorded,and blood samples were taken for determination of serum Aβ40 and Aβ42 concentrations.Results The incidence of POCD was 5％ (in C group),56％ (in IR group) or 22％ (in SR group),and there was no significant difference among the three groups.There were no significant differences in the serum Aβ42 and Aβ40 concentrations after surgery among the three groups.Conclusion The mechanism by which sevoflurane or isoflurane in combination with remifentanil anesthesia results in POCD is not related to the levels of blood Aβ40 or Aβ42 in the elderly patients undergoing abdominal surgery.

The different components of crude mycelium of the predominant endophytic Colletotrichum gloeosporioides of Artemisa annua have been extracted by the methods of acid hydrolysate. We compared the effect of the isolated components on artemisin biosynthesis in hairy root cultures. Therefore, the oligosaccharide elicitor from C. gloeosporioides has been partially purified by column chromatography of Sephadex G25. The isolated oligosaccharide B II (elicitor, MW < 2500) has been revealed to promote the production of artemisinin in Artemisia annua hairy root cultures. When hairy roots of 23-day old cultures (later growth phase) were exposed to the elicitor at 0.4 mg/mL for 4 days, the maximum production of artemisinin reached to 13.51 mg/L, a 51.63% increase over the control. This is the first report on the stimulation of artemisinin production in hairy roots by the oligosaccharide elicitor from an endophytic fungus of A. annua.
Artemisia annua ; drug effects ; growth & development ; metabolism ; Artemisinins ; metabolism ; Bioreactors ; Colletotrichum ; chemistry ; Mycelium ; chemistry ; Oligosaccharides ; isolation & purification ; pharmacology ; Plant Roots ; drug effects ; growth & development ; metabolism

To explore the role of regulatory peptides in the secretion of bronchial epithelial cells (BECs), we observed the effects of four peptides, i.e.vasoactive intestinal peptide (VIP), epidermal growth factor (EGF), endothelin-1 (ET-1), and calcitonin gene-related peptide (CGRP), on the secretion of ILs from unstimulated or O3-stressed BECs. The results of the experiments showed that VIP exerted an inhibitory effect on the secretion of IL-1 and IL-8 from unstimulated and O3-stressed BECs, VIP also decreased the secretion of IL-5 from O3-stressed BECs; EGF promoted secretion of IL-1 and IL-8 from unstimulated BECs, but decreased the secretion of ILs from O3-stressed BECs; ET-1 and CGRP enhanced the secretion of IL-1, IL-5, and IL-8 from unstimlated BECs, CGRP also increased the secretion of ILs from O3-stressed BECs. The results obtained demonstrate that intrapulmonary regulatory peptides modulate the secretion of ILs from BECs, and may play an important part in transduction of inflammatory signals.
Animals ; Bronchi ; cytology ; Calcitonin Gene-Related Peptide ; pharmacology ; Cells, Cultured ; Endothelin-1 ; pharmacology ; Epidermal Growth Factor ; pharmacology ; Epithelial Cells ; drug effects ; secretion ; Female ; Interleukins ; secretion ; Male ; Rabbits ; Vasoactive Intestinal Peptide ; pharmacology

OBJECTIVETo study the effects of a small interfering RNA targeting CXCR-4 (shRNA-CXCR4) on angiogenesis of human breast cancer cells.

METHODSThe expression of CXCR4 mRNA and protein in 3 breast cancer cell lines with CXCR-4 silencing mediated by shRNA-CXCR4 was detected by RT-PCR and Western blotting, respectively. The morphological changes of human umbilical vein endothelial cells (HUVECs) were observed in co-culture with human breast cancer cells after CXCR4 gene silencing.

RESULTSCXCR4 mRNA and protein expressions decreased significantly in MCF-7, MDA-MB-231 and MDA-MB-435s breast cancer cells after the gene silencing (P<0.05). Gene silencing with shRNA-CXCR4 in human breast cancer cells significantly inhibited the ability of HUVECs to form tubular structures in the co-culture (P<0.05).

CONCLUSIONGene silencing by shRNA-CXCR4 can obviously lower the angiogenesis-inducing ability of human breast cancer cells.

Breast Neoplasms ; blood supply ; genetics ; Cell Line, Tumor ; Coculture Techniques ; Endothelial Cells ; cytology ; Female ; Humans ; Neovascularization, Pathologic ; genetics ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Receptors, CXCR4 ; genetics ; metabolism ; Umbilical Veins ; cytology

BACKGROUNDHyaluronidase (Hyase) is an enzyme which hydrolyses hyaluronan (HA), a large nonsulfated glycosaminoglycan. Several genes have been identified to code for hyaluronidases in humans. Its role has only recently been underlined in the invasion of prostate cancer, colonic cancer, and breast cancer. Moreover, the findings were in agreement with some experimental results which showed that HA-derived oligosaccharides had angiogenesis-promoting activity. All these findings prompted us to investigate factors that had been characterized as putative invasive factors in different human breast cancer-derived cell lines.

METHODSWe selected two series of human breast cancer-derived cell lines whose expression of estrogen receptors (ER) was previously published. Hyaluronidase secretion in culture medium and expression of matrix metallo-proteinase (MMP)-9, cathepsin-D (cath-D) and vascular endothelial growth factor (VEGF) by cells were determined. We also investigated cell invasiveness in the Matrigel invasion assay, and studied the capability of cancer cells to promote in vitro formation of tubules by endothelial cells.

RESULTSER(-) cells secreted significantly more hyaluronidase (P < 0.001) and expressed significantly more VEGF (P < 0.01), MMP-9 (P < 0.05) and cath-D (P < 0.0001) than ER(+) cells. Invasion through Matrigel by ER(-) Hyase(+) cells was significantly higher than that by ER(+) Hyase(-) cells (P < 0.05). In both cases, invasion was decreased by heparin (P < 0.05). When ECV-304 endothelial cells were co-cultivated in millicell chambers with cancer cells, ECV-304 cells were induced to form tubules. Tubule formation was demonstrated to be more prominent with ER(-) Hyase(+) cells than with ER(+) Hyase(-) cells (P < 0.05).

CONCLUSIONInvasive features of ER(-) breast cancer cells can be characterized in vitro by an invasive Matrigel assay, as the induction of tubule formation by ECV-304 endothelial cells, higher secretion of hyaluronidase, and higher expression of proteinases MMP-9, cath-D, and the angiogenesis promoting factor VEGF.

Breast Neoplasms ; metabolism ; Cathepsin D ; metabolism ; Cell Line, Tumor ; Humans ; Hyaluronoglucosaminidase ; metabolism ; Immunohistochemistry ; Matrix Metalloproteinase 9 ; metabolism ; Neoplasm Invasiveness ; genetics ; Receptors, Estrogen ; genetics ; Vascular Endothelial Growth Factor A ; metabolism

Objective To establish a Juema minipig model of myocardial infarction, to evaluate the clinical indi?ces in the model pigs, and to explore the relationship between gene expression and metabolic decompensation. Methods 13 male Juema minipigs were randomly divided into control (Sham, n=5), myocardial infarction (MI, n=5) and normal control (for evaluating the recovery condition after surgery, n=3) groups. In the MI group, the ligation was done at the left descending coronary artery around the 1/3 distance to heart apex. Four weeks after the surgery, the cardiac function and serum biochemistry was analyzed. The histological changes and gene expression profiles in the myocardium in the peri?infarct area were exanimated. Results Ultrasonic images showed that the infarction was formed, the ejection fraction and fraction shortening were significantly reduced in the MI group ( ~32% and ~40% less than those of the sham group). Histological examination showed that myocardial fibers at the peri?infarct area were broken, dissolved, and there was con?nective tissue hyperplasia with increased neutrophil and lymphocyte infiltration. Microarray analysis revealed that two myo?cardial remodeling and pathology mediating pathways, three inflammation?related pathways, and 8 metabolic pathways ( in?cluding fatty acid, amino acid, and glucose metabolic pathways) were significantly changed. Conclusions We have suc?cessfully established a Juema minipig model of myocardial infarction. The less branches of the left descending coronary ar?tery allow us to establish a stable model by surgery with comparable characteristics in the clinic indices. The results of this study provides useful reference characteristics of an animal model with characteristic changes in the peri?infarct area.

OBJECTIVETo improve the chemically-activated luciferase expression (CALUX) bioassay for detection of dioxin-like chemicals (DLCs) based on the toxicity mechanisms of DLCs.

METHODSA recombinant vector was constructed and used to transfect human hepatoma (HepG2). The expression of this vector was 10-100 folds higher than that of pGL2 used in previous experiments. The transfected cells showed aromatic hydrocarbon receptor (AhR)-meditated luciferase gene expression. The reliability of luciferase induction in this cell line as a reporter of AhR-mediated toxicity was evaluated, the optimal detection time was examined and a comparison was made by using the commonly used ethoxyresoufin-O-deethylase (EROD) activity induction assay.

RESULTSThe results suggested that the luciferase activity in recombinant cells was peaked at about 4 h and then decreased to a stable activity by 14 h after TCDD treatment. The detection limit of this cell line was 0.11 pmol/L, or 10-fold lower than in previous studies, with a linear range from 1 to 100 pmol/L, related coefficient of 0.997, and the coefficient of variability (CV) of 15-30%.

CONCLUSIONThe luciferase induction is 30-fold more sensitive than EROD induction, the detection time is 68 h shorter and the detection procedure is also simpler.

Biological Assay ; methods ; Carcinoma, Hepatocellular ; pathology ; Cytochrome P-450 CYP1A1 ; biosynthesis ; Environmental Pollutants ; adverse effects ; pharmacology ; Enzyme Induction ; Gene Expression Regulation ; Humans ; Luciferases ; biosynthesis ; Polychlorinated Dibenzodioxins ; adverse effects ; pharmacology ; Sensitivity and Specificity ; Transfection ; Tumor Cells, Cultured

OBJECTIVETo explore the origin and differentiation of gastrointestinal stromal tumors (GISTs).

METHODSImmunohistochemistry staining and electron microscopy were adopted.

RESULTSIn 212 cases of primary GISTs, the positive rates of CD117, CD34, alpha-SMA, MSA, desmin, S-100, PGP9.5 were 96.7%, 77.3%, 19.3%, 15.6%, 1.9%, 16.3%, and 12.3% respectively. Among them, GISTs showed a diffuse and strong positivity for CD117. Electron microscopy of tumor cells demonstrated numerous mitochondria, prominent perinuclear Golgi complex, smooth and rough endoplasmical reticulum and intermediate filaments. Irregular caveolae, dense plaque, incontinuous basal lamina were observed occasionally. Cytoplasmic processes were often observed accompanying with local adhesion present between the processes or between the processes and the cell membrane.

CONCLUSIONSData from both immunophenotype and electron microscopy suggest that GIST might originate from the mesenchymal cells, differentiating to be ICC afterwards, and possessing myoid characteristics in various extent.

Cell Differentiation ; Gastrointestinal Stromal Tumors ; chemistry ; ultrastructure ; Golgi Apparatus ; ultrastructure ; Humans ; Immunohistochemistry ; Microscopy, Electron ; Proto-Oncogene Proteins c-kit ; analysis ; S100 Proteins ; analysis ; Stromal Cells ; chemistry ; ultrastructure ; Ubiquitin Thiolesterase ; analysis

OBJECTIVENucleic acid amplification (PCR) fluorogenic quantitative assay is used for the diagnosis of respiratory syncytial virus (RSV) infection. This study was designed to explore the sensitivity of PCR fluorogenic quantitative assay for ascertaining respiratory RSV infection and RSV infection conditions by detecting the presence of RSV-RNA related sequences in children.

METHODSBronchial and nasopharyngeal secretions specimens from 261 hospitalized children with respiratory tract infections from January 2007 to October 2008 were collected. Respiratory syncytial virus nucleic acid (RNA) in the specimens was measuredby PCR fluorogenic quantitative assay. Blood RSV-IgM was detected by enzyme linked immunosorbent assay (ELISA). The sensitivity for ascertaining respiratory RSV infection was compared between the two assays.

RESULTSThe RSV-RNA positive rate ascertained by PCR fluorogenic quantitative assay (38.7%) was significantly higher than blood RSV-IgM positive rate (21.1%) (p<0.01). The RSV-RNA positive rate (43.6%) in children at ages of less than 6 months was significantly higher than that in children at ages of 1 to three years (32.1%) (p<0.01). The RSV-RNA positive rate in children with bronchiolitis (58.5%) was the highest, followed by bronchopneumonia (38.2%) and acute bronchitis (20.0%).

CONCLUSIONSThe sensitivity of PCR fluorogenic quantitative assay for ascertaining respiratory RSV infection is higher. RSV is a major pathogen of lower respiratory tract infections in infants and young children. A higher rate of RSV infection is associated with a younger age. RSV infection is the most common in children with bronchiolitis.

Antibodies, Viral ; blood ; Child ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Female ; Fluorescence ; Humans ; Immunoglobulin M ; blood ; Infant ; Male ; Polymerase Chain Reaction ; methods ; RNA, Viral ; analysis ; Respiratory Syncytial Viruses ; genetics ; immunology ; isolation & purification ; Respiratory Tract Infections ; virology ; Sensitivity and Specificity ; Sputum ; virology

To enhance immunogenicity of HIV-1 cross neutralizing epitopes , three HIV-1 cross neutralizing epitopes (ELDKWA, NWFDIT, GPGRAFY) were fused to 3' end of HBV S gene by PCR cloning technology, respectively. Three vaccinia virus (Tiantan strain) recombinants expressing separately the three fusion genes were subsequently constructed, named as RVJ1175S-2F5 (ELDKWA), RVJ1175S-4E10 (NWFDIT) and RVJ1175S-447-52D (GPGRAFY), respectively. From the supernatants of CEF cells infected by these vaccinia recombinants, three subunit vaccines (PS-2F5, PS-4E10 and PS-447-52D) were prepared after purification. Biology and immunology characteristics of these fusion antigens in vaccinia recombinants and subunit vaccines were comparatively studied. It was confirmed by PCR and sequencing that the fusion genes were inserted into the TK locus of vaccinia virus (Tiantan strain) correctly. The Fusion proteins were expressed efficiently and secreted into supernatant of the infected cells, which was demonstrated by HBsAg ELISA test. Two typical HBsAg bands of 23kD and 27kD were detected in all the purified samples by SDS-PAGE. These two bands were reacted well to HBsAb and corresponding HIV-1 monoclonal antibodies 2F5, 4E10 and 447-52D. BALB/c mice were immunized with subunit and vaccinia recombinant vaccines by intraperitoneal injection. High levels of HBsAb and anti-HIV-1 cross neutralizing epitope antibody in peripheral blood of immunized mice were tested by ELISA, and all the antibody titers induced by three subunit vaccines were higher than that induced by correlated vaccinia recombinants in mice. This work provides a basis for future study on neutralizing activity of these immunized sera and enhancing immune effect through the combined immunization with different type of vaccines.
AIDS Vaccines ; immunology ; Amino Acid Sequence ; Animals ; Antibody Formation ; Blotting, Western ; Cross Reactions ; Epitopes ; Female ; HIV-1 ; immunology ; Hepatitis B Surface Antigens ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Recombinant Fusion Proteins ; immunology ; Vaccination ; Vaccines, Synthetic ; immunology