The growth and pigment production of 47 strains on casein medium were studied comparatively.T4 and Neurospora crassa AS 3.1602 were selected for their capacity of producing melanin on five different culture media. Melanin produced by T4 was investigated, and T4 was identified as Proteus mirabilis primarily.

Objective To evaluate the influential factors of iron acquisition during Francisella tularensis LVS infection of mouse macrophages.Methods F.tularensis LVS expressing green fluorescent protein was used to infect murine macrophage J774A.1 cells.Transferrin receptor 1(Tfr1)was detected with mono-antibody and visualized with a goat-anti mouse IgG conjugated to Alexa 594.The expression profile of 5 iron metabolism related genes of J774A.1 murine macrophages uninfected or infected with F.tularensis LVS was determined with real-time PCR.Immunoblot analysis was used to compare the Tfr1 expression of live Francisella infected macrophage with dead bacteria.Tfr1 knock-off in J774A.1 cells was performed with siRNA.The transfected cells were infected with Francisella for immunoblotting and microscopy and infection assay.Results It was revealed that the live vaccine strain of F.tularensis induced the expression of Tfr1 in host macrophages.Gene expression analysis indicated that F.tularensis LVS drove an active iron acquisition program with induction of Tfr1 and iron regulatory proteins(Irp1 and Irp2).It was shown by Western-blotting that the siRNA-Tfrc-1 could knock off about 75% of Tfr1 in J774A.1 cells.It was determined by infection assay that,Tfr1 was knocked off,the bacteria number at 1h infection with Francisella was not different from that of control(F=1.06,P=0.326 5),while it was decreased significantly after 24h of infection(F=24.12,P=0.000 6).Conclusions It is demonstrated that upregulation of the Tfr1 may be mediated by post-transcriptional regulation during early infection,but sustained later through increased expression of Irp 1 and Irp 2.Increased expression of Tfr1 expands the intracellular iron pool through transferrin-mediated delivery and may thus be readily available for uptaking by Francisella.Knocking off the expression of Tfr1 does not affect bacterial invasion.Francisella,however,may fail to proliferate in macrophages in which the expression of transferrin receptor has been suppressed.

A white-rot fungi Rigidoporus sp.W-1 which could produce laccase was isolated. The fermentation conditions in shaking flasks were investigated. The optimal carbon source was wheat bran and (NH 4) 2SO 4 was the optimal nitrogen source. The components of the medium were optimized by orthogonal experiment. When W-1 was cultured under the optimum conditions, the activity of laccase could get to 7.1U/mL in 7 days.A great amount of crude laccase could be obtained by adding fresh medium to the 7 days old mycelium.

Multimedia computing technology teaching is a new kind teaching method. It conquers many shortcomings such as poor video, poor expression in traditional teaching of microbiology. But it can also strangle the improvising creation of different teachers in teaching and then lead it to be typical " computer teaching" . So we should obey the teaching discipline in making multimedia computing technology courseware. It can lead many sections part from the class teaching if just emphasize the full use of video and audio. We should make the purpose obviously, and make it as easy as possible in teaching, and we should also explore the fixed regularity to make the teaching and studying integrated perfectly.

Dendritic cells (DCs) derived from bone marrow cells are specialized cells for the uptake, processing, and presentation of foreign and self-antigens. The study indicated that re-transfusion of DCs pulsed with tumor-associated antigen can induce an vigorous specific anti-tumor response in clinic. The present study was aimed to investigate the enhancing effect of DCs derived from human cord blood on T cells in killing tumor cells. Human cord blood mononuclear cells were isolated from human cord blood by density gradient centrifugation using lymphocyte separating medium, and cord blood mononuclear cells were obtained by adherence and cultured in a liquid culture system with GM-CSF and IL-4 for 15 days. Then the cells were analyzed for phenotypes of CD1a by indirect immunofluorescence. The capacity of DCs to initiate T cell-dependent anti-tumor immune responses was assayed by MTT kit. The ratios of DCs to tumor cells in experimental groups were 20:1, 50:1 and 100:1 respectively. The DCs were not added in control group. The results indicated that in the presence of GM-CSF and IL-4, the DCs with typical morphological features at days 15 were observed. At that time, (43.12 +/- 5.83)% CD1a(+) cells were obtained. In addition to these phenotypic properties, the DC of experimental groups could remarkably initiate T cell-dependent anti-tumor immune responses with different ratios compared with control group (P < 0.01), there were no significant difference of killing effects between 100:1 and 50:1 groups (P > 0.05), and killing effect of DC in 20:1 group was higher than that in 100:1 or 50:1 groups (P < 0.05). It is concluded that human cord blood mononuclear cells can serve as a better source of DC, which can promote the capacity to initiate T cell-dependent anti-tumor immune responses.
Apoptosis ; immunology ; Brain Neoplasms ; pathology ; Cell Differentiation ; Cells, Cultured ; Dendritic Cells ; cytology ; immunology ; Fetal Blood ; cytology ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Interleukin-4 ; pharmacology ; Leukocytes, Mononuclear ; cytology ; Neuroblastoma ; pathology ; Recombinant Proteins ; T-Lymphocytes ; immunology ; Tumor Cells, Cultured

OBJECTIVETo evaluate the therapeutic effect of endovascular placement of iodine-125 seed strand and stent combined with transcatheter arterial chemoembolization (TACE) to treat hepatocellular carcinoma (HCC) with tumor thrombus in the main portal vein (MPVTT).

METHODSFifty patients with HCC complicated by MPVTT were enrolled into this study. There were 46 men and 4 women with a mean age of 53.9 years. TACE was performed after the iodine-125 seed strand and self-expandable stent placement in the obstructed segment of the main portal vein (MPV).

RESULTSTechnical success rate was 100% for placement of iodine-125 seed strand and stent in the target segment of MPV. No serious procedure-related complications occurred. The mean follow-up duration was 208.5 d. The mean and median survival time was 370.1 d and 223.0 d, respectively. The 90-, 180-, 360-day cumulative survival rates were 97.5%, 59.3%, and 38.4%, respectively. The mean and median patent time of stent was 524.2 d and 407.4 d, respectively. The 90-, 180-, 360-day cumulative patency rates of stent were 94.9%, 75.2%, and 64.5%, respectively.

CONCLUSIONEndovascular placement of iodine-125 seed strand and stent combined with TACE is an effective therapy for HCC with tumor thrombus in the main portal vein.

Adult ; Aged ; Carcinoma, Hepatocellular ; pathology ; therapy ; Chemoembolization, Therapeutic ; methods ; Combined Modality Therapy ; Female ; Follow-Up Studies ; Humans ; Iodine Radioisotopes ; therapeutic use ; Liver Neoplasms ; pathology ; therapy ; Male ; Middle Aged ; Neoplastic Cells, Circulating ; Portal Vein ; pathology ; Stents ; Survival Rate

5' and 3' flanking region of ovine BLG were amplified from sheep genomic DNA according to the published whole sequence of ovine BLG and cloned to pGEM-T vector correspondently. By partially sequencing, the sequences of BLG 5' and 3' flanking were the same as that of publication completely. The recombinant structure used to direct exogenous gene especially to express in mammary gland was constructed by joining 4.2 kb 5' flanking with 2.1 kb 3' flanking. In order to assess the efficiency of BLG regulatory elements, green fluorescent protein (GFP) gene as a reporter was fused with BLG construct and transfected the mammary epithelial cells (TD47). Through observation under UV microscope and detection by fluorometer, it is demonstrated that the GFP has been successfully expressed in TD47 cell line. By virtue of direct observation and quantitative analysis, the BLG-GFP construct can be served as a model for the quick assessment of mammary gland expression construct.
3' Flanking Region ; genetics ; 5' Flanking Region ; genetics ; Animals ; Breast ; cytology ; Cell Line ; Cloning, Molecular ; Gene Expression Regulation ; Genes, Reporter ; Green Fluorescent Proteins ; Lactoglobulins ; biosynthesis ; genetics ; Luminescent Proteins ; biosynthesis ; genetics ; Sheep

Leber's hereditary optic neuropathy (LHON) is a maternally inherited disorder leading to rapid, painless, bilateral and usually permanent central vision loss in young adults, males are preferentially affected. The maternal transmission of this visual dysfunction in LHON families suggested that mutations in the mitochondrial DNA (mtDNA) are the molecular bases of the disorder. The ND1 G3460A, ND4 G11778A and ND6 T14484C mutations in the genes encoding the subunits of respiratory chain complex I, account for more than 50% of LHON families worldwide. These three mutations are designated to be primary mutations because they impart a high risk for LHON expression. However, matrilineal relatives within and among families, despite carrying the same LHON-associated mtDNA mutation(s), exhibit a wide range of onset, severity, and the progression of visual impairment. These findings strongly indicated that the LHON-associated primary mutation(s) are the primary factors underlying the development of vision loss, but they themselves are insufficient to produce a clinic phenotype. The prone to male, incomplete penetrance, and phenotypic variability of vision loss suggest that other modifier factors including personal factors, environmental factors, nuclear modifier genes and mitochondrial haplotypes contribute to the phenotypic expression of these mtDNA mutations. In particular, the mitochondrial haplotypes may play a synergic role in the development of vision loss in the families carrying the LHON-associated primary mtDNA mutation(s).
DNA, Mitochondrial ; genetics ; Genome, Human ; genetics ; Genomics ; Haplotypes ; Humans ; Mitochondria ; genetics ; Optic Atrophy, Hereditary, Leber ; genetics ; pathology

Three-dimensional (3D) culture of cells could closely mimic the in vivo situation with regard to cell function and microenvironment compared with plane monolayer cultured cells. In this paper, we established 3D culture of rat WB-F344 cells with rotary cell culture system (RCCS) to simulate microgravity environment, and examined cells proliferation, morphology, microstructure, E-cadherin protein quantity and mRNA expression of adhesion molecules by count the number of cells, optical microscope, transmission electron microscope and reverse transcriptase-polymerase chain reaction (RT-PCR). The results demonstrated that cells were polyhedron with lots of micovilli and mitochondria, which grow well and packed together densely to form irregular aggregates. Adjacent cells were connected with desmosome and tight junction. With the regard, the aggregates behaved 3D growth characteristics. Moreover, compared with control, mRNA level of Fibronectin and E-cadherin protein were increased, the changes maybe is the part mechanism in this microgravity simulated cells culture models which strengthened cells junction. This rotating 3D model might facilitate the study of interactions of cell-cell, cell-matrix and the mechanisms.
Animals ; Cadherins ; genetics ; Cell Adhesion ; Cell Culture Techniques ; methods ; Cell Proliferation ; Fibronectins ; genetics ; Rats ; Spheroids, Cellular ; ultrastructure ; Weightlessness Simulation

OBJECTIVETo explore the mechanism of tau hyperphosphorylation and the effect of LiCl on tau phosphorylation and the memory retention deficits in streptozotocin-induced diabetes mellitus (DM) rats.

METHODSThe rats were randomly divided into control, DM, DM + NaCl, and DM + LiCl groups and diabetes was induced by streptozotocin. The activity of glycogen synthase kinase-3 (GSK-3) was measured by 32P-labelling. The level of tau phosphorylated and changes of memory retention were examined by Western blotting and step down test, respectively.

RESULTSCompared with control group, the activity of GSK-3 and tau phosphorylation was increased, and the memory retention was impaired in DM group. When the rats were treated with LiCl, the activity of GSK-3 and hyperphosphorylation of tau were significantly arrested (P < 0.05, P < 0.01), and the memory retention deficit was significantly improved (P < 0.05).

CONCLUSIONThe hyperphosphorylation of tau can be induced by activation of GSK-3 in diabetic rats. Lithium protects tau from hyperphosphorylation and may rescue memory retention in the rats by inhibiting GSK-3 activity.

Animals ; Cerebral Cortex ; metabolism ; Diabetes Mellitus, Experimental ; complications ; Glycogen Synthase Kinase 3 ; metabolism ; Lithium Chloride ; therapeutic use ; Male ; Memory Disorders ; drug therapy ; metabolism ; Phosphorylation ; drug effects ; Rats ; Rats, Sprague-Dawley ; tau Proteins ; metabolism