Acute Disease ; Adult ; Benzene ; poisoning ; Female ; Humans ; Leukemia ; chemically induced ; Occupational Exposure ; adverse effects

Through mechanical analisis,it is discovered that the twined structure decides the amplification coefficient of the catgut tension.Many troubles of production can be explained with this mechanics model,and it could be used as a reference for medical sulures. 　　The paper gives an accurate calculating formular for catgut design and its direct tensile strength through improving mechanics model. It can raise the material strength from 10 to over 31% depending on its twined structure.

This study was to improve the way for selecting ura5 mutants of Cryptoccocus neoformans Cap59 capsule-deficient strains.They were induced by Diethyl Sulfate. Ura5 mutants were screened by 5-fluoroorotic acid counter selection method. Using the new method, we obtained two ura5 mutants of Cryptoccocus neoformans Cap59 capsule-deficient strain.A easy method that was used to screen ura5 mutants of Cryptoccocus neoformans has been established.

OBJECTIVESBased on the immunologic character of Pichia pastoris yeast, a new therapeutic vaccine, whole recombinant yeast, was used to explore a new way to activate cell-mediated anti-viral immunity.

METHODSThe recombinant plasmids, pPIC9K/S and PIC9K/hsp(1-370)-S, were constructed by inserting the gene encoding HBsAg, HSP70 (1-370) -HBsAg into vector pPIC9K and then the recombinants were transfected into Pichia pastoris yeast,GS115, respectively. Then that recombinant yeast immunized BALB/C mice were detected for humoral and cellular immunity to HBsAg.

RESULTSRecombinant yeast successfully activated the humoral immunity to HBsAg in mice, but failed to activate the cellular immunity.

CONCLUSIONThe whole recombinant yeast can be used as vaccine, but need further study for optimal way of immunization.

Animals ; Female ; HSP70 Heat-Shock Proteins ; genetics ; immunology ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; genetics ; immunology ; Hepatitis B Vaccines ; immunology ; Mice ; Mice, Inbred BALB C ; Pichia ; genetics ; Plasmids ; T-Lymphocytes, Cytotoxic ; immunology ; Vaccines, Synthetic ; immunology

OBJECTIVETo evaluate the application of fluorescence in-situ hybridization (FISH) in detection of gene translocation in paraffin-embedded tissue samples of synovial sarcoma.

METHODSInterphase FISH was carried out in paraffin-embedded tissue of 42 cases of synovial sarcoma and 9 cases of non-synovial sarcoma, using a LSI SYT (18q11.2) dual color break-apart probe. In all of the cases studied, the gene fusion product SYT-SSX was also analyzed by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSPositive signals were detected in 37 cases (88.1%) of synovial sarcoma by FISH, as compared with 35 cases (83.8%) by RT-PCR and 39 cases (92.9%) by both techniques. Of the 39 positive cases, 33 cases (78.5%) revealed SYT gene translocation.

CONCLUSIONSFISH may serve as an adjunctive diagnostic tool in problematic cases of synovial sarcoma and can be applied in paraffin-embedded tissue samples. As compared with RT-PCR, FISH is also sensitive and reliable. The methodology is less labor intensive and time consuming. FISH has great potential in molecular diagnosis of soft tissue tumors.

Adolescent ; Adult ; Biomarkers, Tumor ; genetics ; Child ; Child, Preschool ; Chromosome Aberrations ; Female ; Head and Neck Neoplasms ; genetics ; metabolism ; Humans ; In Situ Hybridization, Fluorescence ; Lower Extremity ; pathology ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; Paraffin Embedding ; Reverse Transcriptase Polymerase Chain Reaction ; Sarcoma, Synovial ; genetics ; metabolism ; Soft Tissue Neoplasms ; genetics ; metabolism ; Young Adult

Acetyl-N-glucosaminyltransferase gene (nodC) was successfully cloned to Escherichia coli from Mesorhizobium loti. The recombinant E. coli harboring nodC gene was able to synthesize chitooligosaccharides (COs) in MMYNG medium. In optimized condition, a yield of 526 mg/L was obtained after 26 h cultivation in 10 L bioreactor. COs concentration reached up to 4.5% of the cell dry weight. The COs products were purified by charcoal adsorption and Bio-gel P4 chromatography, penta-N-acetylchitopentaose (m/z, 1034[M + H]+) and tetra-N-acetylchitopentaose (m/z, 831 [M + H]+) were identified as the dominating COs product using the method of liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS).
Bacterial Proteins ; genetics ; metabolism ; Chitosan ; isolation & purification ; metabolism ; Chromatography, Gel ; Chromatography, High Pressure Liquid ; Escherichia coli ; genetics ; metabolism ; N-Acetylglucosaminyltransferases ; genetics ; metabolism ; Oligosaccharides ; biosynthesis ; isolation & purification ; Recombinant Proteins ; metabolism ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo observe the status of AKT and phospho-AKT (pAKT) in three diffuse large B cell lymphoma (DLBCL) cell lines, and to investigate the effects of AKT activation on biologic behavior of DLBCL cells.

METHODSThree DLBCL cell lines, ly1, ly8 and ly10 were maintained in 10% FBS or serum free culture medium. The expression of AKT and status of pAKT were detected by Western blotting. LY294002, an inhibitor of PI3K, was used to suppress the level of pAKT. Flow cytometry combined with PI staining, AnnexinV-FITC assay and Brdu incorporation assay were used to analyze the parameters of the cell cycle, apoptosis and proliferation respectively.

RESULTSThere was constitutive activation of AKT in three DLBCL cell lines and the levels of pAKT were altered in the different environments. In 10% FBS culture medium, pAKT was higher than that in serum free culture medium in ly8 and ly10, however, pAKT in ly1 maintained in serum free culture medium was mildly higher than that in 10% FBS culture medium. When the cell lines ly1, ly8, ly10 were maintained in 10% FBS culture medium, the inhibitor LY294002 suppressed the level of pAKT efficiently in three DLBCL cell lines. The percentage of cells at S phase and the proliferation index were significantly decreased (P < 0.05) without an increase of apoptosis (P > 0.05).

CONCLUSIONSActivation of AKT may play an important role in the development of DLBCL. It is closely related to the control of cell cycle and proliferation, but is not associated with apoptosis. LY294002 can inhibit cell growth by decreasing the levels of pAKT in DLBCL cell lines.

Apoptosis ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chromones ; pharmacology ; Culture Media, Serum-Free ; Enzyme Activation ; Humans ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; Morpholines ; pharmacology ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors ; Phosphorylation ; drug effects ; Proto-Oncogene Proteins c-akt ; metabolism

OBJECTIVECT120B gene is a splicing variant of CT120A, which deletes 96 nucleotides and leads to an in-frame loss of 32 amino acids between the codon 136 and 167 as compared with CT120A. This study was undertaken to assess the effects of CT120B expression on lung cancer cell growth and to explore the gene expression profiles.

METHODSCT120B cDNA was transfected into the human lung adenocarcinoma SPC-A-1 cells, and stable cell lines overexpressing CT120B were established. CCK-8 assay and tumorigenecity in a xenograft model were performed to analyze cell proliferation in vitro and in vivo. The differential gene expression induced by overexpressed CT120B was investigated using Atlas cDNA expression array. Flow cytometry was performed to analyze cell cycle and cell apoptosis.

RESULTSOverexpression of CT120B in SPC-A-1 cells resulted in a reduced cell growth rate in vitro, and decrease of the tumorigenicity in nude mice. A total of 38 genes were identified as differential expressions with more than a 2.0-fold change by Atlas cDNA expression array analysis, including downregulated cyclin E1, cdk 2, c-kit, CXCR4 and upregulated caspase 8 gene. Overexpression of CT120B also induced G1 phase arrest, but had no effect on cell apoptosis.

CONCLUSIONThe G1 cell cycle arrest, but not apoptosis, underlay the growth inhibitory activities of CT120B. The down-regulation of c-kit and CXCR4 expression might also contribute to the suppressive effects on cell growth of CT120B.

Adenocarcinoma ; metabolism ; pathology ; Animals ; Cell Line, Tumor ; Cell Proliferation ; DNA, Complementary ; genetics ; G1 Phase ; Gene Expression Profiling ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Male ; Membrane Proteins ; genetics ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Proteins ; genetics ; metabolism ; Neoplasm Transplantation ; Oligonucleotide Array Sequence Analysis ; Proto-Oncogene Proteins c-kit ; metabolism ; Receptors, CXCR4 ; metabolism ; Transfection

OBJECTIVETo explore whether bone marrow stem cells (MSCs) from adult mice can be induced to differentiate into hepatocytes by hepatocyte growth factor (HGF) alone and the time phase characteristics in the differentiation progress.

METHODSAdult mouse MSCs were treated with or without 100 ng/ml HGF, on days 0, 7, 14, 21, and 28. The morphologic characteristics of the cells were examined; the albumin (ALB), AFP mRNA was analyzed sub-quantively using reverse transcription polymerase chain reaction (RT-PCR) and immumohistochemistry techniques. The expression of ALB, AFP and CK19 were detected by using anti-ALB, AFP and CK19 antibodies.

RESULTSFreshly isolated adult mouse MSCs expressed ALB and AFP mRNA weakly; in the group without HGF, no ALB mRNA was detected on day 7. The expression of AFP mRNA was reduced significantly on day 7, and could not be detected anymore after day 14. In the HGF treated group, ALB mRNA was not detected on day 7, but the positive lane appeared again on day 14, and the expression of ALB mRNA was increased on day 21 but reduced in the following days. The AFP mRNA was positive at all times, however it tended to decrease after day 14 in the HGF treated groups. The result of immumohistochemistry was consistent with that of RT-PCR, and CK19 was always negative.

CONCLUSIONAdult mouse MSCs can be induced into hepatocyte differentiation in vitro. The optimal time for the induction was 2 to 3 weeks.

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Hepatocyte Growth Factor ; pharmacology ; Hepatocytes ; cytology ; Male ; Mice ; Stem Cells ; cytology ; Time Factors

OBJECTIVETo detect beta-catenin mRNA levels in sporadic colorectal cancers (SCRC) and adjacent normal colorectal mucosa, and to investigate the association between the beta-catenin mRNA level and its aberrant expression and clinicopathological parameters.

METHODSThe concentration of beta-catenin mRNA in 81 SCRCs and 28 adjacent normal colorectal mucosa specimens was determined by TaqMan real-time quantitative RT-PCR. The ratio of beta-catenin cDNA copies/GAPDH cDNA copies was used to represent the mRNA expression level in different tissues. The beta-catenin protein expression was determined by the EnVision two-step immunohistochemical method.

RESULTSbeta-catenin mRNA levels in SCRCs (2.527 +/- 2.284) were lower than those in the adjacent normal colorectal mucosa (5.003 +/- 3.326), P < 0.05. In addition, beta-catenin mRNA levels in lymph node-positive cases and tumors with ulcerative and infiltrating growth types were significantly lower (1.827 +/- 1.288, 2.202 +/- 2.035) than those in lymph node-negative cases and polypoid growth type tumors (3.359 +/- 2.881, 3.108 +/- 2.610), P < 0.05. No significant difference of beta-catenin mRNA level was found between cases with aberrant beta-catenin cytoplasm or nuclear expression and those without.

CONCLUSIONSSCRCs express lower levels of beta-catenin mRNA than normal colorectal mucosa. Such lower level expression is associated with lymph node metastasis and tumors with ulcerative and infiltrative growth pattern. Aberrant cytoplasmic and nuclear expression of beta-catenin appears unrelated to the lower mRNA levels. Quantitative detection of beta-catenin mRNA may be a useful approach to monitor the biological behavior of SCRCs.

Adenocarcinoma ; genetics ; metabolism ; pathology ; Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; genetics ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Intestinal Mucosa ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; beta Catenin ; biosynthesis ; genetics