人尿中沙丁胺醇的测定

采用GC—Ms联用技术，对尿中的沙丁胺醇(salbutamol)进行分析。实验表明，口服一片沙丁胺醇(2．4 mg)，可于24 h从尿中排泄，且以游离形式为主。样品提取时，控制溶液pH为8.8时，提取率较高。本方法对沙丁胺醇定性简单、快速、准确。     

New Insights on Adenovirus as Vaccine Vectors

Adenovirus (Ad) vectors were initially developed for treatment of genetic diseases. Their usefulness for permanent gene replacement was limited by their high immunogenicity, which resulted in rapid elimination of transduced cells through induction of T and B cells to antigens of Ad and the transgene product. The very trait that excluded their use for sustained treatment of genetic diseases made them highly attractive as vaccine carriers. Recently though results showed that Ad vectors based on common human serotypes, such as serotype 5, may not be ideal as vaccine carriers. A recently conducted phase 2b trial, termed STEP trial, with an AdHu5-based vaccine expressing antigens of human immunodeficiency virus 1 (HIV-1) not only showed lack of efficacy in spite of the vaccine''s immunogenicity, but also suggested an increased trend for HIV acquisition in individuals that had circulating AdHu5 neutralizing antibodies prior to vaccination. Alternative serotypes from humans or nonhuman primates (NHPs), to which most humans lack pre-existing immunity, have been vectored and may circumvent the problems encountered with the use of AdHu5 vectors in humans. In summary, although Ad vectors have seen their share of setbacks in recent years, they remain viable tools for prevention or treatment of a multitude of diseases.     

Purkinje cell degeneration elevates eupneic and hypercapnic ventilation in rats

Previous studies have demonstrated that among cerebellar nuclei, the fastigial nucleus (FN) plays a major role in facilitation of respiration, especially during hypercapnia. Since the FN primarily receives inhibitory afferents from Purkinje cells (PCs), we hypothesized that degeneration of PCs would increase both eupneic and hypercapnic ventilation. Experiments were carried out on 20 animals (n=10 for both normal and PC-degenerated) that were divided into three groups based on the different preparations used, i.e., four pairs for the awake, three pairs for the anesthetized, and three other pairs initially for the awake and subsequently for the anesthetized. The awake normal and PCD rats were separately placed in an unrestrained whole-body plethysmograph and ventilatory parameters measured before (room air) and during hypercapnia (5% CO₂ + 21% O₂ + 74% N₂) for 30 min. The anesthetized animals were exposed to the same level of hypercapnia applied for ∼5 min. The results showed that both eupneic breathing and hypercapnia-induced ventilatory augmentation were significantly greater in the awake PCD rat than those observed in the normal one, primarily due to a remarkable elevation in V_T with little changes in f. The same results were also observed in anesthetized preparations. A Fos protein immunocytochemical approach was employed to determine the effect of degeneration on PCs and FN neuronal activity. Fos expression of PCs was very intensive in normal rats, but absent or diminished in PCD rats. In sharp contrast, FN Fos expression was obscure in normal rats, but very apparent in PCD rats. These data suggest that PCs play an inhibitory role in modulation of eupneic and hypercapnic ventilation via inhibiting FN neuronal activity.     

Understanding inadequate pain management in the clinical setting: the value of the sequential explanatory mixed method study

Aim. The purpose of this paper is to critically explore the sequential explanatory mixed method research design and how it can enhance our understanding of pain management. Background. The general prevalence of pain after surgery has not changed significantly over several decades despite the widespread introduction of new pain relieving technologies. The majority of postoperative pain studies use quantitative methods which offer little understanding of the underlying processes of care. Understanding can be illuminated by using an explanatory mixed method research design. Design. Discursive paper. Method. This paper focuses on the methodological considerations when using a mixed method design. Two previously published mixed methods studies illustrate how findings can inform practice. In the first, 85 women undergoing surgery completed questionnaires to measure pain, anxiety and depression. Telephone interviews explored their pain experiences. The second study considered frequency and patterns of anxiety in the immediate pre and postoperative period. Semi‐structured telephone interviews, identified contributing events/situations amenable to nursing intervention. Discussion. Reasons for growing popularity, criticisms, paradigmatic considerations and epistemological roots of pragmatism are explored. The two explanatory mixed method studies provide examples of these studies and how ‘inferences’ from quantitative and qualitative data can inform practice. Conclusion. This paper connects quantitative and qualitative data, drawing on two research studies, to give greater understanding to the management of pain. Knowledge of the processes responsible for inadequate pain management can be illuminated by using explanatory mixed methods research designs. Relevance to clinical practice. Nursing requires knowledge which reflects the complexity of human health. The explanatory mixed method study can elucidate the problem under scrutiny, e.g. prevalence of pain or anxiety. The qualitative phase can generates an understanding of contributing factors and insights for care delivery. The implicit desire to change and influence practice makes it relevant for those closely aligned to practice.     

NF-kB signaling blockade abolishes implant particle-induced osteoclastogenesis.

In this study we investigated the effect of NF-kB signaling blockade on polymethylmethacrylate (PMMA) particle-induced osteoclastogenesis in vitro. We first established effective blockade of NF-kB activity as tested by electrophoretic mobility shift assays (EMSA). Particle-induced NF-kB activation in murine osteoclast precursor cells (CSF-1-dependent bone marrow macrophages) was markedly reduced by co-treatment of the cells with the NF-kB inhibitors N-tosyl-L-phenylalanine chloromethyl ketone (TPCK) and Calpain Inhibitor I (CPI). This inhibition of NF-kB activity was associated with blockade of p50 NF-kB subunit nuclear translocation. We then established a direct NF-kB inhibition approach by utilizing a TAT-bound, mutant IkB (TAT:IkB(46-317)), and demonstrated an inhibitory effect evidenced by decreased NF-kB DNA binding activity. Having established that these strategies (TPCK, CPI, TAT: IkB(46-317)) effectively block NF-kB activation, we next investigated the effect of these agents on particle-stimulated osteoclast formation. PMMA particle stimulation of mature osteoclast formation from RANKL-primed osteoclast precursor cells was blocked by all three inhibitors. To further test the efficacy of NF-kB blockade, experiments were performed with the TAT:IkB(46-317) mutant peptide in whole bone marrow cultures that contain supporting stromal cells. Again, this inhibitor efficiently blocked particle-induced osteoclastogenesis. Thus, we have shown that pharmaceutical and molecular blockade of NF-kB activation inhibits PMMA particle-directed osteoclastogenesis in vitro.     

HeartMate vented-electric left ventricular assist system: technique for intrathoracic or intraperitoneal implantation via a left thoracotomy.

The HeartMate vented-electric left ventricular assist system (Thoratec Corp., Woburn, MA) has become widely accepted as a temporary bridge to transplantation. We describe a left thoracotomy technique in 3 patients for implanting this pump intrathoracically or intraperitoneally. In all 3 cases, long-term pump function was satisfactory. For HeartMate implantation, the left thoracotomy approach may be particularly useful when previous median sternotomies, coupled with the severe debilitation posed by chronic heart failure and hepatic dysfunction with resultant coagulopathy, would greatly increase the mortality and morbidity of a redo median sternotomy.     

Regression of fibrosis and hypertrophy in failing myocardium following mechanical circulatory support.

BACKGROUND: The cellular and structural changes that occur during long-term ventricular unloading leading to cardiac recovery are poorly understood. However, we have previously demonstrated that left ventricular assist device (LVAD) support leads to a significant decrease in intracardiac tumor necrosis factor-alpha (TNF-alpha), a protein capable of producing hypertrophy and fibrosis. METHODS: To further define the beneficial effects of long-term ventricular unloading on cardiac function, we determined the effect of mechanical circulatory support on fibrosis and hypertrophy in paired myocardial samples of 18 patients with end-stage cardiomyopathy obtained at the time of LVAD implantation and removal. RESULTS: We determined total collagen as well as collagen I and III by a semiquantitative analysis of positive immune-stained areas in pre- and post-LVAD myocardial samples. We found that total collagen content was reduced by 72% (p < 0.001), whereas collagen I content decreased by 66% (p < 0.001) and collagen III content was reduced by 62% (p < 0.001). Next, we determined myocyte size by direct analysis of cellular dimensions utilizing a computerized edge detection system in pre- and post-LVAD myocardial samples. We found that myocyte size decreased in all patients studied for an average reduction of 26% (33.1 +/- 1.32 to 24.4 +/- 1.64 microm, p < 0.001). CONCLUSION: These data demonstrate that long-term mechanical circulatory support significantly reduces collagen content and decreases myocyte size. We suggest that the reduction of fibrosis and hypertrophy observed may in part contribute to the recovery of cardiac function associated with long-term mechanical circulatory support.