A model for screening the yeast which can ferment xylose to produce the ethanol was constructed.An ethanol yeast was obtained using the lignocellulose as substrate production the ethanol.By malt extract medium pre-culturing,soil samples use the plate with xylose as sole carbon source as the primary screening,then finally screen by the potassium dichromate color-displaying method.A strain named Y2-3 was screened from the soil.Phenotypic analysis including morphology and physiology and biochemical characteristics and 26D1/D2 sequence analysis were carried out.Based on taxonomy results,the Y2-3 was identified as Pichia caribbica.The strain Y2-3 ferments using xylose as sole carbon source: biomass 23.5 g/L,xylose utilization rate 94.7 %,ethanol final yield 4.57 g/L;using mixture sugar:biomass 28.6 g/L,xylose utilization rate 94.2 %,glucose utilization rate 95.6%,ethanol final yield 20.6 g/L.Pichia caribbica is a yeast which can utilize xylose and mixture sugar as substrate.It established the foundation for further research fermentation of ethanol by yeast using lignocellulose.

Objective To investigate the effects of in situ venous arterialization on extensive artery obliterans occlusion of the lower extremity. Methods Lumbar sympathetic ganglionectomy and one stage in situ arterialization of the great saphemous vein were performed in 104 ischemic limbs of 88 patients with extensive arterial occlusion. Results Eighty-two of 104 limbs were followed-up from 6 months to over 6 years. The intermittent claudication, night pain improved in all cases, with satisfactory wound healing and no swelling of the lower limbs. Conclusions Arterial blood flow through venous conduit improves and reconstructs the blood circulation of the ischemic limbs.

In order to observe the proliferative and apoptotic situation of megakaryocytes in patients with myelodysplastic syndromes (MDS). CD41 immunoenzyme labeling (alkaline phosphatase anti-alkaline phosphatase APAAP)/DNA in situ end labelling (ISEL) double stained techniques was used onto plastic cold embedded bone marrow sections in 29 MDS patients to analyse the proliferative and apoptostic characterization of megakaryocytic line with 14 cases of iron deficient diseases (IDA) as control. The results showed that the mean CD41 positive cell number in MDS group was (26.23 +/- 8.18) /mm(2) with a count of (15.64 +/- 7.11) /mm(2) in control group (p < 0.05). The small-micro megakaryocytes in MDS is much higher than that in IDA group (P<0.01). There was a positive co-relation between total megakaryocytes and small-micro megakaryocytes count in MDS (r = 0.702, p<0.01). Some megakaryocytes distributed abnormally around trabecula and formed small or large clusters. Apoptotic megakaryocytes in MDS occupied 4.40% and 9.32% of all CD14 positive cells and all apoptotic cells respectively (p > 0.5 comparing with control). Apoptosis in megakargocytic line occurred only in small-micro megakaryocytes and showed positive co-relation to the number of micro-megakaryocytes. Some apoptotic cell with morphologic characters of megakaryocytes expressed no CD41. It is concluded that overproliferation of megakaryocytes exists in MDS. Apoptosis occurring in micro-megakaryocytes may be a kind of physiological response to abnormal megakaryopoicsis in MDS. No obvious increased apoptosis of megakaryocytes in MDS was found perhaps due to lack of surface antigens CD41 in some later stages of apoptotic cell.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Apoptosis ; Cell Division ; Child ; Female ; Humans ; Male ; Megakaryocytes ; pathology ; Middle Aged ; Myelodysplastic Syndromes ; pathology

We use direct-write laser micromachining technology to fabricate the microfluidic chip, and to establish a microfluidic chemiluminescence immunoassay system based on superparamagnetic microbeads, for detecting alpha- fetoprotein (AFP). The AFP analysis can be completed in 20 minutes with 5 microl sample and 5 microl reagent, and there is a good linear correlation in the range of 1-800 ng/ml.
Equipment Design ; Humans ; Immunoassay ; instrumentation ; methods ; Luminescent Measurements ; instrumentation ; methods ; Microfluidic Analytical Techniques ; instrumentation ; alpha-Fetoproteins ; analysis

OBJECTIVETo develop a rapid and simple multiplex polymerase chain reaction (PCR) method which discriminates extended-spectrum beta-lactamases (ESBLs) genes in sporadic Shigella isolates from 1998 to 2007 in Hangzhou city, China.

METHODSAfter ESBLs screening according to the Clinical and Laboratory Standards Institute (CLSI) method, CTX-M, TEM, SHV and OXA-1 encoding genes were detected by using a multiplex PCR method, and the results were verified by 8 single gene PCR amplification.

RESULTSSeventeen isolates harbored ESBLs genes among 195 Shigella isolates (8.72%). Genes encoding CTX-M (17 strains), TEM (2 strains), OXA-1 (10 strains) and SHV (0 strains) were discriminated with multiplex PCR analysis, which coincided with eight single gene PCR analysis at 94.12%.

CONCLUSIONMultiplex PCR should be a suitable tool for initial rapid screening and discriminating ESBLs genes in Shigella isolates. With similar trend of national surveillance data, the proportion of sporadic Shigella isolates harbouring ESBLs genes might probably be on increase.

DNA, Bacterial ; analysis ; Genes, Bacterial ; Genotype ; Humans ; Microbial Sensitivity Tests ; Polymerase Chain Reaction ; methods ; Shigella ; genetics ; isolation & purification ; beta-Lactamases ; genetics

OBJECTIVETo study the mechanism involved in the control of glioma cell proliferation with transfection of connexin (Cx) 43 gene.

METHODSC6 rat glioma and TJ905 human glioblastoma cell lines without Cx43 gene expression were transfected with Cx43cDNA mediated by lipofectamine. Northern blot, in situ hybridization and immunohistochemical technology were used to detect the expression of Cx43mRNA and its protein with MTT assay and silver colloid stain for the detection of cell proliferation, TUNEL method for determination of cell apoptosis, scrape loading dye transfer (SLDT) for GJIC, Western blot and immunohistochemical technology for bFGF, PDGF, EGFR, IGF-I and IGFBP3 expression.

RESULTSCx 43 gene transfected glioma cells showed decreased proliferation, restored GJIC and decreased bFGF, PDGF, IGFBP3, except EGFR expression and cell apoptosis which showed no change.

CONCLUSIONThe mechanism of Cx 43 gene inhibiting gliomas cell proliferation is the restoration of GJIC and decreased autocrine growth factors.

Animals ; Apoptosis ; Cell Division ; physiology ; Connexin 43 ; genetics ; physiology ; DNA, Complementary ; genetics ; Glioma ; pathology ; Rats ; Transfection ; Tumor Cells, Cultured

OBJECTIVETo investigate the influence of SEPT7 on biological characters of gliomas cells TJ905.

METHODSRecombinant SEPT7 constructs was transfected to human glioblastoma cell line TJ905 in which SEPT7 expression is absent. The positive clones were identified by RT-PCR and Western blot analysis. The cell proliferation was determined by MTT assay and flow cytometry, cell apoptosis was detected with Annexin V staining and cell invasion was evaluated by motility in three-dimensional culture. Moreover, the molecules regulating the cell cycle progression were examined by immunofluorescence staining and Western blot analysis.

RESULTSWhen SEPT7 was successfully transfected to TJ905 cells, the cell proliferation activity of TJ905 cell was inhibited, the cell cycle was arrested in G0/G1 phase and S phase fraction (SPF) was lowered, the positive regulatory molecules for cell cycle progression including cyclin D1, CDk4, cyclin E and CDk2 were downregulated while the negative modulators including p16 and p21 were upregulated, apoptotic cells were increased and cell invasive ability was attenuated.

CONCLUSIONSTransfection of SEPT7 construct into the glioma cells TJ905 is able to inhibit the proliferation activity and invasive ability of TJ905 cell and to induce cell apoptosis. These results revealed that SEPT7 exerted the suppressive effect on the glioma cell growth and invasion, and induced apoptosis, and suggested that SEPT7 as a gene of glioma suppressor.

Apoptosis ; Blotting, Western ; Brain Neoplasms ; genetics ; metabolism ; pathology ; Cell Cycle ; Cell Cycle Proteins ; genetics ; metabolism ; physiology ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival ; Flow Cytometry ; Fluorescent Antibody Technique ; Glioma ; genetics ; metabolism ; pathology ; Humans ; Reverse Transcriptase Polymerase Chain Reaction ; Septins ; Transfection

BACKGROUNDHIV-1 infected and immune-activated macrophages and microglia secrete neurotoxins, such as tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), which play major role in the neuronal death. It has been shown that different HIV-1 variants have varying abilities to elicit secretion of TNF-α by peripheral blood mononuclear cell (PBMC); however, whether the difference of gp120 gene could affect the secretion of TNF-α and IL-1β by glial cells is unknown. The aim of this study was to explore the association between gene diversity and induction of neurotoxic cytokines.

METHODSIn this study, we constructed retroviral vectors MSCV-IRES-GFP/gp120 using HIV-1 gp120 genes isolated from four different tissues of one patient who died of AIDS dementia complex (ADC). Recombinant retroviruses produced by cotransfection of MSCV-IRES-GFP/gp120, pCMV-VSV-G and pUMVC into 293T cells were collected and added into U87 glial cells. Concentrations of TNF-α and IL-1β secreted by transduced U87 cells were assayed with ELISA separately.

RESULTSThe four HIV-1 gp120 were in the different branch of the neighbor-joining tree. Compared to the pMIG retrovirus (gp120-negative) or U87 cells, all the gp120-positive recombinant retroviruses induced more TNF-α (P < 0.01) and IL-1β (P < 0.01). In addition, compared with the L/MIG retrovirus, all the three brain gp120-positive recombinant retroviruses induced less TNF-α (P < 0.01) and IL-1β (P < 0.01).

CONCLUSIONSHIV-1 gp120 could induce U87 cells secret more TNF-α and IL-1β again. The more important is that difference of HIV-1 gp120, especially cell-tropism may account for the different ability in eliciting secretion of TNF-α and IL-1β, which might supply a novel idea helping understand the pathogenesis of ADC.

AIDS Dementia Complex ; metabolism ; virology ; Cell Line ; Cell Line, Tumor ; Enzyme-Linked Immunosorbent Assay ; HIV Envelope Protein gp120 ; genetics ; metabolism ; Humans ; Interleukin-1beta ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo study the anti-invasion effect of SEPT7 gene on U251MG glioma cells and its possible molecular mechanism.

METHODSRecombinant adenovirus vector carrying SEPT7 gene (rAd5-SEPT7) was transduced to human glioma cell line U251MG, and empty adenovirus vector was used as control. Tumor invasion was examined by Transwell method and 3 D-Matrigel assay, and tumor cell migration by wound-healing method and 2 D-Matrigel assay. Three major molecular events associated with cell motility and migration, including changes of expression in MMP2, MMP9, MT1-MMP, TIMP1 and TIMP2, the alteration of integrin alpha(v)beta(3) expression, and the structural change of cytoskeleton protein, tubulin-alpha, in U251 cells transduced with rAd5-SEPT7 were studied by Western blotting, immunofluorescence and laser scanning confocal microscope, respectively.

RESULTSThe invasive and migratory capabilities of cells transduced with rAd5-SEPT7 were inhibited. The expression of extracellular matrix metalloproteinases MMP-2, MMP-9, MT1-MMP and integrin alpha(v)beta(3) was significantly decreased, while the expression of matrix metalloproteinase inhibitor TIMP1, TIMP2 was upregulated. Intracellular cytoskeleton protein-tubulin-alpha in U251 cells exhibited prominent morphological changes which including the appearance of distortion and aggregation resulting from redistribution of tubulin-alpha, and this feature of alteration was similar to the tubulin-alpha structure in normal non-tumor cells.

CONCLUSIONSEPT7 gene can inhibit the invasion and migration ability of U251 glioma cells. Its molecular mechanism may include that SEPT7 gene reverses the imbalanced state of MMPs/TIMPs, downregulates the expression of integrin alpha(v)beta(3) and alters the structure of tubulin-alpha of U251MG glioma cells. It is suggested that SEPT7 gene could be a good candidate for gene therapy of gliomas.

Adenoviridae ; genetics ; Blotting, Western ; Cell Cycle Proteins ; genetics ; physiology ; Cell Line, Tumor ; Cell Movement ; genetics ; Genetic Vectors ; genetics ; Glioma ; metabolism ; pathology ; Humans ; Integrin alphaVbeta3 ; metabolism ; Matrix Metalloproteinase 14 ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Microscopy, Confocal ; Neoplasm Invasiveness ; genetics ; Septins ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism

OBJECTIVETo study the correlation between DNA fingerprinting of Mycobacterium tuberculosis (MTB) stains isolated from the Chinese army in the south and from local residents, and to investigate the molecular epidemiological characteristics of tuberculosis (TB) in the army, for the sake of TB prevention in the army.

METHODSMTB DNA was digested with restriction endonuclease PvuII and electrophoresed in agarose gel, after Southern Blotting, the membrane was hybridized with a 245 bp fragment of IS6110 which labeled [alpha(32)P]-dCTP as probe. Finally, a restriction fragment length polymorphism (RFLP) patterns was shown, and analyzed logestic with epidemiological data from the patients.

RESULTSA total number of 185 TB strains were detected and the IS6110 copy numbers ranged from 1 - 22. No significant difference was found in the IS6110 copy numbers between patients from army and local patients. IS6110 copy numbers of TB strains in army patients were centered in 6 - 20, however, with 7 - 20 copies in local TB patients. The TB strains were dispersed into 8 groups and the majority of TB strains in both army and local patients was centered in groups I, II, III. The distribution of DNA fingerprint for drug resistance TB strains was significantly different from those for sensitive strains. No different distribution of among groups was found regarding BCG history.

CONCLUSIONSThe genetics of TB stains were roughly the same between the army patients and local ones, but there was a strong correlation in the gene levels. Data suggested that a close connection should be considered on TB prevention and treatment for TB patients in the army and local residents.

China ; epidemiology ; DNA Fingerprinting ; DNA, Bacterial ; analysis ; genetics ; Humans ; Military Personnel ; Molecular Epidemiology ; Mycobacterium tuberculosis ; genetics ; isolation & purification ; Polymorphism, Restriction Fragment Length ; Tuberculosis ; epidemiology ; genetics ; microbiology