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Cream, Carlos L., Aihua Li, and Eugene E. Nattie. RTNTRH causes prolonged respiratory stimulation. J. Appl.Physiol. 83(3): 792-799, 1997.We injectedthyrotropin-releasing hormone (TRH; 10 nl; 0.25, 0.5, 1.0, or 10 mM),its inactive free acid form (TRHOH; 1 mM), or a metabolite with lowTRH-receptor binding affinity, histidine-proline diketopiperazine (cHP;1 mM), into the retrotrapezoid nucleus of anesthetized rats. Injectionlocation was verified by anatomic analysis. Lower doses (0.25-0.5mM) significantly increased both the product of integrated phrenicamplitude and frequency(Phr · f) and f for 20-30min compared with artificial cerebrospinal fluid control injections. Higher doses (1.0-10 mM) produced greater and long-lastingstimulation of Phr · f,Phr, and f and of blood pressure. Thisstimulation reached values 150% of baseline and durations of 270 minafter a single injection. TRHOH (1 mM ) or cHP (1 mM) had no effect onPhr but increased f, as did 1 mM TRH. We concludethat TRH has a very powerful stimulatory effect in the retrotrapezoidnucleus region on Phr · f, withthe Phr response seemingly specific for TRHreceptors. Similar responses of f to TRHOH and cHP suggest it may benonspecific.

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Cells disseminated from primary epithelial tumors into peripheral blood, called circulating tumor cells (CTCs), can be monitored to assess metastases and to provide a surrogate marker of treatment response. Here, we demonstrate how the flexible micro spring array (FMSA) device—a novel microfluidic device that enriches CTCs by two physical parameters: size and deformability—could be used in the rational development of treatment intervention and as a method to study the fundamental biology of CTCs. Cancer cells of different origins were spiked into healthy samples of donor blood to mimic blood samples of metastatic cancer patients. This spiked human blood was filtered using the FMSA device, and the recovered cells were successfully expanded in vitro and in a novel in vivo system. A series of experiments were performed to characterize these cells and to investigate the effect of chemotherapy on the resulting cultures. As few as 20 colon cancer cells in 7.5 mL blood could be isolated with the FMSA device, expanded both in vitro and in vivo and used at 25 cells per well to obtain significant and reliable chemosensitivity data. We also show that isolating a low number of viable patient CTCs and maintaining them in culture for a few weeks is possible. The isolation of viable cancer cells from human blood using the FMSA device provides a novel and realistic means for studying the biology of viable CTCs and for testing drug efficacy on these rare cells—a hypothesis that can be tested in future clinical trials.  相似文献   
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Thyrotropin-releasing hormone (TRH) injected into the retrotrapezoid nucleus (RTN) of anesthetized rats produces a large, prolonged stimulation of ventilatory output (C. L. Cream, A. Li, and E. E. Nattie. J. Appl. Physiol. 83: 792-799, 1997). Here we inject or dialyze TRH into the RTN of conscious rats. In 6 of 17 injections (200 nl, 3.1 +/- 1.7 mM), ventilation (VE) increased 31% by 10 min, with recovery by 60 min. With dialysis, each animal of one group (n = 5) received, in random order, 10 mM TRH, 10 mM TRHOH (a metabolite of TRH), and artificial cerebrospinal fluid (aCSF); each animal of a second group (n = 5) received aCSF and 1 mM TRH. TRHOH and aCSF had no sustained effects. TRH (1 mM) increased VE (32%, P < 0.02, by 10 min, with recovery by 60 min), O(2) consumption (VO(2); 19%, P < 0. 03), and body (rectal) temperature (T(re); 0.5 degrees C, P < 0.09). TRH (10 mM) increased VE (78%, P < 0.01, by 10 min, with no recovery at 60 min), VO(2) (48%, P < 0.01), and T(re) (1.0 degrees C, P < 0. 01). TRH also induced arousal. The tissue volume affected in dialysis, estimated by spread of dialyzed fluorescein (332.3 mol wt, mol wt of TRH = 362.4), was 1,580 +/- 256 nl for 10 mM (n = 5) and 590 +/- 128 nl for 1 mM (n = 5). We conclude that 1) the RTN is involved in the integration of VE, VO(2), T(re), and arousal and 2) TRH may establish the responsiveness of RTN neurons.  相似文献   
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