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61.
Yamamoto N Yang M Jiang P Xu M Tsuchiya H Tomita K Moossa AR Hoffman RM 《Clinical & experimental metastasis》2003,20(7):633-638
We have established stable, bright green fluorescent protein (GFP)- or red fluorescent protein (RFP)-expressing HT-1080 human
fibrosarcoma clones. These cell lines showed similar cell proliferation rates and high-frequency experimental lung metastasis.
The HT-1080-GFP and -RFP clones enable simultaneous real-time dual-color imaging in the live animal. HT-1080 cells were transduced
with retroviral vectors containing GFP or RFP and the neomycin resistance gene. Stable transformants were selected stepwise
with G418 up to 800 μl/ml. Subsequently, high GFP- or RFP-expressing clones, HT-1080-GFP or HT-1080-RFP, respectively, were
selected. 3×106 cells from each clone were mixed and injected into the tail vein of SCID mice. The cells seeded the lung at high frequency
with subsequent formation of pure green and pure red colonies as well as mixed yellow colonies with different patterns visualized
directly on excised lungs. The lung metastases were also visualized by external fluorescence imaging in live animals through
skin-flap windows over the chest wall. Lung metastases were observed on the lung surface of all mice. SCID mice well tolerated
multiple surgical procedures for direct-view imaging via skin-flap windows. Real-time metastatic growth of the two different
colored clones in the same lung was externally imaged with resolution and quantification of green, red, or yellow colonies
in live animals. The color coding enabled determination of whether the colonies grew clonally or were seeded as a mixture
with one cell type eventually dominating, or whether the colonies grew as a mixture. The simultaneous real-time dual-color
imaging of metastatic colonies described in this report gives rise to the possibility of color-coded imaging of clones of
cancer cells carrying various forms of gene of interest.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
62.
Here we present the changes in cortical activity occurring within a few hours after a 1-h exposure to a 120-dB SPL pure tone (5 or 6 kHz). The changes in primary auditory cortex of 16 ketamine-anesthetized cats were assessed by recording, with two 8-microelectrode arrays, from the same multiunit clusters before and after the trauma. The exposure resulted in a peripheral threshold increase that stabilized after a few hours to on average 40 dB in the frequency range of 6-32 kHz, as measured by the auditory brain stem response. The trauma induced a shift in characteristic frequency toward lower frequencies, an emergence of new responses, a broadening of the tuning curve, and an increase in the maximum of driven discharges. In addition, the onset response after the trauma was of shorter duration than before the trauma. The results suggest the involvement of both a decrease and an increase in inhibition. They are discussed in terms of changes in central inhibition and its implications for tonotopic map plasticity. 相似文献
63.
Culture of chondrocytes in fibroin-hydrogel sponge 总被引:5,自引:0,他引:5
Aoki H Tomita N Morita Y Hattori K Harada Y Sonobe M Wakitani S Tamada Y 《Bio-medical materials and engineering》2003,13(4):309-316
Fibroin-hydrogel sponge and collagen gel were used as scaffold for in vitro cartilage regeneration. Fibroin-hydrogel sponge was formed by phase separation from freezed fibroin solution. Chondrocytes were harvested from proximal humerus, distal femur and proximal tibia of 4-week-old Japanese white rabbits and inoculated in the fibroin-hydrogel sponge and collagen gel. Those constructs were cultured in DMEM supplemented with 10% FCS and 50 ml L-ascorbate at 37 degrees C. Histological observation, measurement of sulfated glycosaminoglycan and cell density were carried out at 3, 7, and 14 days after the cultivation. Well-defined cartilage tissue can be seen both in the fibroin-hydrogel sponge and in the collagen gel. The matrix was intensely stained by safranin-O and showed a metachromatic reaction in both group. However, the quantity of sulfated glycosaminoglycan and cell density of the fibroin-hydrogel sponge group were increased more rapidly than these of the collagen gel group. Thus, the chondrocytes proliferated in the fibroin sponge without losing their differentiated phenotype. It is possible that culture environment in the fibroin sponge was suitable for chondrocytes regeneration. 相似文献
64.
An autosomal dominant posterior polar cataract locus maps to human chromosome 20p12-q12 总被引:4,自引:0,他引:4
Yamada K Tomita H Yoshiura K Kondo S Wakui K Fukushima Y Ikegawa S Nakamura Y Amemiya T Niikawa N 《European journal of human genetics : EJHG》2000,8(7):535-539
We assigned the locus for a previously reported new type of autosomal dominant posterior polar cataract (CPP3) to 20p12-q12 by a genome-wide two-point linkage analysis with microsatellite markers. CPP3 is characterized by progressive, disc-shaped, posterior subcapsular opacity. The disease was seen in 10 members of a Japanese family and transmitted in an autosomal dominant fashion through four generations. We obtained a maximum lod score (Zmax) of 3.61 with a recombination fraction (theta) of 0.00 for markers D20S917, D20S885 and D20S874. Haplotype analysis gave the disease gene localization at a 15.7-cM interval between D20S851 and D20S96 loci on chromosome 20p12-q12. Since the BFSP1 that encodes the lens-specific beaded filament structural protein 1 (filensin) has been mapped around the CPP3 region, we performed sequence analysis on its entire coding region. However, no base substitution or deletion was detected in the CPP3 patients. The mapping of the CPP3 locus to 20p12-q12 not only expands our understanding of the genetic heterogeneity in autosomal dominant posterior polar cataracts but also is a clue for the positional cloning of the disease gene. 相似文献
65.
66.
Rabbit factor P, which is involved in stabilizing the labile C3 convertase of the alternative complement pathway, was isolated from rabbit serum by a simple two-step procedure: polyethylene glycol precipitation and QAE-Sephadex A-50 column chromatography. The chemical properties, and functional activities of the rabbit P were very similar to those of human P, except that there were slight differences in the SDS-PAGE patterns of limited tryptic digests and in the amino terminal sequences. The molecular weights were estimated to be 58 000 for rabbit P and 59 000 for human P on SDS-PAGE under reducing conditions. The amino terminal 36 residues of rabbit P showed 78% homology to the equivalent region of human P. 相似文献
67.
Body-oriented therapies as relaxation training and certain forms of meditation are gaining popularity in the treatment and prevention of psychosomatic disorders. In this paper, a new method of self-control called self-regulation method (SRM), derived from autogenic training and Zen meditation, is presented. The technique of SRM is introduced. Secondly, physiological studies on SRM using skin temperature, galvanic skin response, and cortical evoked potentials are presented. Thirdly, the results of psychological tests conducted on SRM are presented. These psycho-physiological studies suggest that SRM may elicit a state of 'relaxed alertness'. Fourthly, clinical applications of SRM are discussed, and 3 cases are presented. Finally, SRM is discussed in relation to the psychology and physiology of 'relaxed alertness'. 相似文献
68.
Kuniaki Tanaka Yoshifumi Tomita Mari Tsuruta Fumiko Konishi Masao Okuda Kunisuke Himeno Kikuo Nomoto 《Immunopharmacology and immunotoxicology》1990,12(2):277-291
Chlorella vulgaris, an unicellular green algae, or its acetone-extract (Ac-Ex) were administered orally to Meth A tumor bearing BALB/c or (BALB/c DBA/2)F1 (CDF1) mice. When CDF1 mice were fed daily with 10% dried powder of Chlorella vulgaris (CVP) containing diet before and after Meth A tumor inoculation, the growth of rechallenged Meth A tumor was significantly suppressed in an antigen-specific manner. Augmentation of antitumor resistance was exhibited also by Winn assay using lymph node cells of tumor-bearing mice orally administered with CVP or Ac-Ex. Antigen-specific concomitant immunity in these mice were mediated by cytostatic T cells but not by cytotoxic T cells. Natural killer cells seemed not to contribute in antitumor resistance in this system. 相似文献
69.
70.
BACKGROUND: Our aims were to follow the longitudinal changes in lumbar spine bone mineral density (BMD) with multiple pregnancies, and to study whether polymorphisms in the vitamin D receptor (VDR) and estrogen receptor (ER) genes may influence the results. METHODS: We repeatedly measured the BMD of the lumbar spine (L2-L4) of 133 women who had undergone two successive pregnancies and 73 non-pregnant controls, and analysed the restriction fragment length polymorphisms using restriction endonucleases TaqI, ApaI and FokI for the VDR gene, and PvuII and XbaI for the ER gene. RESULTS: Cases and controls had no significant differences in the longitudinal BMD changes. The mean percentage change in lumbar BMD (DeltaBMD%) of the women with the XX/Xx genotype was significantly lower than that of the women with the xx genotype after adjusting for age at each delivery, BMD of the first scan, and interval between the scans (0.2 +/- 3.3 versus 2.0 +/- 4.2%; P=0.030, analysis of covariance). Multiple regression analyses to evaluate the contribution of the XbaI polymorphism of the ER gene on DeltaBMD% showed that the percentage decrease in BMD was greater for women lacking the XbaI restriction site (adjusted R2=0.188, P<0.0001). CONCLUSIONS: The present study suggests that the DeltaBMD% was significantly influenced by the XbaI polymorphism of the ER gene. 相似文献