Aneurysm, False ; pathology ; Carotid Artery, Internal ; pathology ; Humans ; Male ; Sphenoid Sinus ; pathology ; Young Adult

Objective To evaluate Cica-Beta Test kit for detection of metallo-β-lactamase-producing Pseudomonas aeruginosa (PAE)in the clinical microbiology laboratory.Methods A total of 82 imipenem-resistant PAE clinical isolates from litera-ture[5]was dectected to metallo-β-lactamase (MBLs)by PAE-MHT and Cica-Beta Test kit.Results The sensitivity,speci-ficity and accuracy rate of PAE-MHT was 84.6%,97.2% and 97.6%,and the sensitivity,specificity and accuracy rate of Cica-Beta Test kit was 76.9%,100% and 96.3%,respectively.Two methods had a good consistency.Conclusion Two methods are simple,quick for detecting to metallo-β-lactamase-producing Pseudomonas in clinical laboratories.

Cryptosporidium andersoni ATP-binding cassette (CaABC) is an important membrane protein involved in substrate transport across the membrane. In this research, the nucleotide binding domain (NBD) of CaABC gene was amplified by PCR, and the eukaryotic expression vector of pEGFP-C1-CaNBD was reconstructed. Then, the recombinant plasmid of pEGFP-C1-CaNBD was transformed into the mouse intestinal epithelial cells (IECs) to study the iron transportation function of CaABC. The results indicated that NBD region of CaABC gene can significantly elevate the transport efficiency of Ca2+, Mg2+, K+, and HCO3 - in IECs (P<0.05). The significance of this study is to find the ATPase inhibitors for NBD region of CaABC gene and to inhibit ATP binding and nutrient transport of CaABC transporter. Thus, C. andersoni will be killed by inhibition of nutrient uptake. This will open up a new way for treatment of cryptosporidiosis.
ATP-Binding Cassette Transporters/*chemistry/*genetics/metabolism ; Adenosine Triphosphate/metabolism ; Amino Acid Sequence ; Animals ; Calcium/metabolism ; *Cloning, Molecular ; Cryptosporidiosis/parasitology ; Cryptosporidium/chemistry/genetics/*metabolism ; Humans ; Iron/metabolism ; Mice ; Molecular Sequence Data ; Protein Structure, Tertiary ; Protozoan Proteins/*chemistry/*genetics/metabolism ; Sequence Alignment

OBJECTIVETo trace the embryonic stem (ES) cells transplanted into rat brain by labeling the cells with green fluorescent protein (GFP) and by mouse neuronal specific antibody Thy-1 and compare their features.

METHODSFor GFP labeling,transfect pEGFP-N1 plasmid containing GFP and anti-neomycin sequences into embryonic stem cell and add neomycin for more than 10 passages. To test the GFP expression in vivo, the GFP-ES was transplanted into healthy rat brain, and the frozen sectioned slides were observed under fluorescence microscope and laser con-focal microscope 21 days later. For the antibody labeling,embryonic stem cells were directly transplanted into the rat brain. The specific mouse thy-1 antibody was used in immunostaining of transplanted cells. For both of the two labeling method, the slides were also examined by double labeling with the antibodies,neuronal nuclei (NeuN) or glial fibrillary acidic protein (GFAP) to identify the differentiation of transplanted cells.

RESULTSBoth single ES cell and cell pellets expressed bright green fluorescence the day after plasmid transfection, and more than 30% ES cells were labeled. The GFP-labeled cells could still be found gathered around the infusion channel at least 21 days later, but the GFP fluorescent could not be overlapped with NeuN or GFAP staining. On the contrary, Thy-1 antibody overlapped well with NeuN or GFAP staining.

CONCLUSIONSLiposome-helped plasmid GPF transfection is effective in labeling mouse embryonic stem cell in vivo,but is not effective in showing the differentiated cells. On the contrary, Thy-1 antibody can not only show the transplanted cells, but also trace the transplanted cells after their differentiation.

Animals ; Cell Differentiation ; physiology ; Cells, Cultured ; Embryonic Stem Cells ; cytology ; physiology ; transplantation ; Green Fluorescent Proteins ; Male ; Mice ; Mice, Transgenic ; Rats ; Rats, Sprague-Dawley ; Staining and Labeling ; methods

The study was aimed to observe the effect of recombinant human granulocyte-colony stimulating factors (rhG-CSF) in low dose on peripheral blood stem cell (PBSC) mobilization in unrelated healthy normal donors. G-CSF was administered at 5 microg/(kg x d) subcutaneously for successive 5 or 6 days to 56 unrelated donors. Stem cells were harvested on the fourth and fifth days or on the fifth and sixth days. The numbers of mononuclear cells (MNC), CD34(+) cells and Hb, Plt, and CD3(+), CD4(+), CD8(+) and CD20(+) cells were determined during the mobilization. The results showed that most common adverse events were bone pain (17.9%, 10/56), agrypnia (8.9%, 5/56) and lassitude (4.5%, 3/56) during rhG-CSF mobilization, but all donors were suffered less than grade II according to the WHO criteria, and did not need to stop the mobilization and not need to give special treatment. In harvest on day 4 - 5 and 5 - 6, MNC count was (5.95 +/- 1.52) x 10(8)/kg and (7.19 +/- 2.12) x 10(8)/kg; CD34(+) cells count was (3.03 +/- 1.09) x 10(6)/kg and (7.92 +/- 2.50) x 10(6)/kg. There were no significant differences in hemoglobin level and platelet count, the percentage of CD3(+) cells, CD4(+) cells, CD8(+) cells and CD20(+) cells between pre-mobilization and post-mobilization of rhG-CSF. It is concluded that the low dose of rhG-CSF 5 microg/(kg x d) for peripheral blood stem cell mobilization in unrelated healthy normal donors is safe and effective.
Adolescent ; Adult ; Blood Donors ; Female ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Recombinant Proteins

OBJECTIVETo study the chemical constituents of the aerial parts of Liusticum chuanxiong.

METHODThe chemical components were isolated by silica gel and Sephadex LH-20 column chromatography. Structures were elucidated on the basis of physico-chemical properities and spectral data.

RESULTEight chemical constituents were isolated, and identified as protocatechuic acid (1), caffeic acid (2), scopoletin (3), apigenin (4), quercetin (5), cosmosiin (6), kaempferol-3-O-beta-D-glucopyranosid (7) and glucose (8).

CONCLUSIONCompounds 1-8 were obtained from the aerial parts of the plant for the first time, compounds 3-8 were obtained from the plant for the first time.

Apigenin ; chemistry ; isolation & purification ; Kaempferols ; chemistry ; isolation & purification ; Ligusticum ; chemistry ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry ; Scopoletin ; chemistry ; isolation & purification

OBJECTIVETo compare the influences of stapedectomy and small fenestra stapedotomy on the hearing of guinea pigs.

METHODSTwenty-four (48 ears) guinea pigs were randomized equally into two groups, and the left ears were subjected to stapedectomy and total stapes replacement with a prosthesis, or sham operation (12 ears) to expose the footplate of the stapes and the round window. Each guinea pig was tested by ABR perioperatively. Four guinea pigs were chosen randomly from each group and decapitated for morphological examination by light microscopy and scanning electron microscopy after ABR test.

RESULTSIn the sham operation group, the post-operative latencies of each wave, the intervals and the hearing threshold exhibited no significant changes other than prolonged latency of wave I. In stapedectomy group, the hearing threshold increased to 23.75-/+3.77 dBSPL 1 h after operation with significantly prolonged post-operative latencies of all the waves and intervals but for III-IV interval, which was shortened. The latencies of each wave (especially waves I and III) in the stapedectomy group were increased by a greater magnitude than those in the sham operation group, but the intervals were comparable between the two groups. No significant difference was noted in the parameters of ABR either 1 h or 1 day after the operation between the two groups, in which the architecture of cochleas remained intact with similar number of spiral ganglion cells. The stereocilia of the outer hearing cells (OHC) were normal in the sham operation group while in stapedectomy group, slight stereocilia disorder occurred but became normal 1 day after operation. No obvious changes were found in the stereocilia of the inner hearing cell (IHC) in either groups.

CONCLUSIONStapedectomy can induce mild hearing loss without seriously damaging the function of the cochlea in guinea pigs.

Animals ; Auditory Threshold ; physiology ; Evoked Potentials, Auditory, Brain Stem ; physiology ; Female ; Guinea Pigs ; Hair Cells, Auditory ; cytology ; Hearing ; physiology ; Male ; Round Window, Ear ; cytology ; physiology ; surgery ; Stapes Surgery ; adverse effects ; Time Factors

OBJECTIVETo explore anti-angiogenesis effect of Mer, a member of tyrosine kinase receptor family, and its mechanism.

METHODSHuman Mer full length plasmid was transfected into HMEC-1 cells through liposome. G418 was used to select positive clone. Expression of Mer at mRNA and protein level was detected by real-time PCR and Western-blot, respectively. Transwell and Matrigel were used to evaluate the effect of overexpressed Mer on migration and angiogenesis of HMEC-1 cells. Primary angiogenesis associated factor VEGF-A, VEGF-B, VEGF-C, VEGF-D and VEGFR-1, VEGFR-2 were screened by real-time PCR.

RESULTSAfter G418 selection, the Mer expression in transfected HMEC-1 cells was increased 3.61- and 2.12 fold at mRNA and protein level, respectively. Compared with negative control, the migration of Mer-HMEC-1 was decreased (21 +/- 6 vs 36 +/- 11), and angiogenesis capability on Matrigel significantly decreased. By real-time PCR, the expression of VEGF-C and VEGFR-2 was down-regulated to 44.7% and 25.6% of the negative control.

CONCLUSIONOverexpressed Mer tyrosine kinase receptor can inhibit the migration and angiogenesis of HMEC-1 cells through VEGF-C/VEGFR-2 signal pathway.

Cell Line ; Cell Movement ; Cell Proliferation ; Endothelial Cells ; metabolism ; physiology ; Endothelium, Vascular ; cytology ; Humans ; Neovascularization, Physiologic ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Receptor Protein-Tyrosine Kinases ; genetics ; metabolism ; Receptors, Vascular Endothelial Growth Factor ; metabolism ; Transfection ; Vascular Endothelial Growth Factors ; metabolism ; c-Mer Tyrosine Kinase

BACKGROUNDNumerous studies have shown that reducing the level of tumor necrosis factor-alpha (TNFα) through the use of anti-TNF antibodies or soluble TNF receptor is a safe and efficacious treatment to inflammatory diseases such as rheumatoid arthritis. Therefore, novel approaches to achieve this outcome are desired. The aim of this study was to investigate the characterization of a small molecule inhibitor, Y316, which blocks TNF mRNA upregulation and TNF production by lipopolysaccharides (LPS) stimulated monocytes.

METHODSPeripheral blood mononuclear cells (PBMC) from healthy volunteers were plated in 24-well plates and stimulated with LPS (1 µg/ml), phorbol-12-myristate-13-acetate (PMA) (100 ng/ml), zymosan (10 µg/ml) and Tsst (100 ng/ml). Supernatants were collected after 4-hour culture at 37°C, and quantitative determination of TNFα, interleukin-1β (IL-1β), IL-6, IL-8, IL-10 and IL-2 production in the supernatants was performed by colorimetric enzyme-linked immunosorbent assay (ELISA). Total RNA of PBMC was isolated and cytokine mRNA quantitation was performed by using a RNA level measuring kit (R & D Systems). PBMC were pretreated with Y316 (10 µmol/L, 1 µmol/L, 0.1 µmol/L, 0.01 µol/L and 0.001 µmol/L) or dimethyl sulfoxide at 37°C for 10 minutes, and then stimulated with LPS or PMA, protein concentrations of p44.42, IKBα, P38 and Jun NH2-terminal kinase were determined by Western blotting. Cyclic adenosine-3',5'-monophosphate (cAMP) of PBMC was measured by enzyme immunoassay kit (Amersham Pharmacia Biotech).

RESULTSY316 blocked TNF production and inhibited the upregulation of TNF mRNA levels in response to LPS, and also prevented the production of IL-1 and IL-6. In contrast, Y316 augmented the production of IL-10 in LPS-stimulated monocytes. Y316 failed to prevent the production of IL-2 and TNF in antigen-stimulated T cells, suggesting that its effects may be cell-type specific. Y316 prevented the phosphorylation and activation of the MAPK, ERK, and therefore appeared to mediate its effects on TNF by acting at an early point in the signaling cascade induced in response to LPS. There was no effect of Y316 on cAMP levels either alone or in the presence of LPS.

CONCLUSIONSY316 appears to be a small molecule inhibiting TNF production, which may act via a novel mechanism. Identification of the target of Y316 may lead to the development of alternative strategies for achieving selective cytokine inhibition.

Anti-Inflammatory Agents ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Humans ; Interleukin-1 ; antagonists & inhibitors ; biosynthesis ; Interleukin-6 ; antagonists & inhibitors ; biosynthesis ; Lipopolysaccharides ; pharmacology ; Monocytes ; drug effects ; immunology ; Phosphorylation ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors ; biosynthesis