HPLC法测定卡巴拉汀贴剂中药物含量及其体外透皮释放量

建立了一种卡巴拉汀贴片含量及其体外透皮释放量的高效液相色谱(HPLC)测定法,并用离体豚鼠皮肤为透皮屏障,采用改进Franz扩散池,研究卡巴拉汀透皮贴剂体外透皮性能.结果表明,卡巴拉汀在9.69～116.32μg/mL浓度范围内线性关系良好,检测限为0.17 μg/,mL,精密度RSD为0.12%～0.47%,低、中、高3个浓度的平均回收率分别为98.79%、98.59%、99.88%;该贴剂体外透皮释放曲线符合零级方程(Q=kt),为Q=59.91t+6.02,r=0.9987.所建立的HPLc方法灵敏、准确、操作简便,可有效测定卡巴拉汀贴剂的含量及其体外透皮释放量.     

Improvement of sexual function in testosterone deficient men treated for 1 year with a permeation enhanced testosterone transdermal system

PURPOSE: The effects of androgen replacement via a nonscrotal permeation enhanced testosterone transdermal system on the sexual function of men with hypogonadism were assessed. MATERIALS AND METHODS: An open label, multicenter study of testosterone supplementation and withdrawal was conducted with sexual function assessed by the Watts and Davidson questionnaires and RigiScan monitoring. RESULTS: When comparing results obtained during use of the testosterone transdermal system (with normalized testosterone levels) and during the androgen withdrawal period, nocturnal erections occurred more frequently with longer duration and greater rigidity, and patient assessments of sexual desire and weekly number of erections were higher. CONCLUSIONS: Sexual function improved significantly in men with hypogonadism treated with the testosterone transdermal system.     

Transient increase in circulating donor leukocytes after allogeneic transfusions in immunocompetent recipients compatible with donor cell proliferation

Donor leukocytes in therapeutic blood components are implicated in transfusion-related complications ranging from alloimmunization to graft-versus-host disease (GVHD) to viral transmission and reactivation. To further characterize the kinetics of donor leukocyte clearance after allogeneic transfusion, we developed allele-specific polymerase chain reaction (PCR) assays directed at a single-copy Y chromosome gene and HLA class II alleles. These assays enable sensitive detection and quantitation of donor leukocytes at concentrations ranging from one cell to greater than 1,000 cells per 125 microL of recipient blood. When applied to serial samples from five consecutive orthopedic surgery patients who met study criteria, we observed 99.9% clearance of donor leukocytes over the initial 2 days posttransfusion, followed by a transient, 1-log increase in circulating donor leukocytes on days 3 to 5. This phenomenon was reproduced in a canine transfusion model, where the transient donor leukocyte expansion phase was prevented by gamma irradiation of donor blood, and was not observed after transfusions into alloimmunized dogs. We hypothesize that this transient increase in circulating allogeneic donor cells represents one arm of an in vivo mixed lymphocyte reaction, with activated donor T lymphocytes proliferating in an abortive GVHD reaction to HLA-incompatible recipient cells. Further investigation of this phenomenon should provide insight into the mechanisms involved in donor-recipient leukocyte interactions posttransfusion and the relationship of these interactions to leukocyte-induced complications.     

Comparison of agar dilution, broth microdilution, E-test, disk diffusion, and automated Vitek methods for testing susceptibilities of Enterococcus spp. to vancomycin

An evaluation was undertaken to determine the optimal method for testing the susceptibilities of 100 clinical isolates and two reference strains of Enterococcus spp. to vancomycin in vitro. Six testing methods were studied by using the following media and incubation times: agar screen with the Synergy Quad Plate (Remel, Lenexa, Kans.), an in-house-prepared brain heart infusion (BHI) agar plate, and an in-house-prepared Mueller-Hinton (MH) agar plate, all incubated for 24 or 48 h; broth microdilution (Sensititre Just One Strip; AccuMed International, Inc., West Lake, Ohio) with BHI or cation-adjusted MH broth incubated for 24 or 48 h; agar dilution with BHI or MH agar incubated for 24 or 48 h; epsilometer test (E test; AB BioDisk, Solna, Sweden) with BHI or MH agar incubated for 24 or 48 h; disk diffusion with BHI or MH agar incubated for 24 or 48 h; and the automated Vitek method with the gram-positive susceptibility Staphylococcus aureus card and R02.03 software (bioMerieux, Inc., Hazelwood, Mo.). Growth failures occurred with MH media (n = 6) but not with BHI media. One growth failure occurred with the Vitek method. Results for each testing method for each Enterococcus strain were interpreted as susceptible, intermediate, or resistant according to current National Committee for Clinical Laboratory Standards (NCCLS) criteria and compared to the vancomycin resistance genotype (i.e., vanA, vanB, vanC-1, or vanC-2/3). For all methods, extension of the incubation time from 24 h to 48 h either produced no difference in the results or gave poorer results. The following methods produced no very major or major interpretive errors: broth microdilution with BHI media incubated for 24 h, agar dilution with BHI media incubated for 24 or 48 h, and E test with BHI media incubated for 24 or 48 h. Unacceptable frequencies of very major errors (> 1%) occurred with all methods for which MH media were used. Minor interpretive errors were frequent with all methods. These minor interpretive errors also occurred most frequently with Enterococcus strains with vanC genes, which encoded low-level vancomycin resistance (MIC < or = 8 microg/ml), as opposed to Enterococcus strains which possessed vanA or vanB genes, which encoded higher-level vancomycin resistance (MIC > or = 64 microg/ml). Modification of NCCLS breakpoints, especially for motile Enterococcus spp. (E. casseliflavus, E. flavescens, and E. gallinarum), may resolve this problem; however, in the current study, one E. faecalis strain and one E. faecium strain carried only the vanC gene. The agar screen method may also require reformulation. The current agar screen plate contains 6 microg of vancomycin per ml, which may not detect all low-level resistance associated with vanC genotypes. Nevertheless, the clinical significance of this low-level vancomycin resistance remains unknown.     

Electrophoretic gels of dentin matrix proteins as diffusion media for in vitro mineralization

Non-collagenous proteins of dentin and bone have important effects on mineralization which have been studied by various in vitro systems. We developed an in vitro mineralization system using electrophoretic gels as diffusion media of calcium and phosphate ions. Calcium and phosphate ions were diffused naturally or propelled by electric potential. Calcium phosphate was precipitated in the gel, and the precipitation was affected by proteins in the gel which had been separated by electrophoresis. We applied this system to analysis of non-collagenous proteins of dentin. Among the proteins, phosphophoryns promoted calcium phosphate precipitation in the natural-diffusion system. A non-collagenous protein having a molecular mass of 60 kDa inhibited precipitation. The results were different, however, in the electric-diffusion system, in which phosphophoryns had a negative effect. The present system enabled us to compare the effects of plural proteins rapidly, even using unpurified material.     

Strategies for rare-event detection: an approach for automated fetal cell detection in maternal blood

This article explores the feasibility of the use of automated microscopy and image analysis to detect the presence of rare fetal nucleated red blood cells (NRBCs) circulating in maternal blood. The rationales for enrichment and for automated image analysis for "rare-event" detection are reviewed. We also describe the application of automated image analysis to 42 maternal blood samples, using a protocol consisting of one-step enrichment followed by immunocytochemical staining for fetal hemoglobin (HbF) and FISH for X- and Y-chromosomal sequences. Automated image analysis consisted of multimode microscopy and subsequent visual evaluation of image memories containing the selected objects. The FISH results were compared with the results of conventional karyotyping of the chorionic villi. By use of manual screening, 43% of the slides were found to be positive (>=1 NRBC), with a mean number of 11 NRBCs (range 1-40). By automated microscopy, 52% were positive, with on average 17 NRBCs (range 1-111). There was a good correlation between both manual and automated screening, but the NRBC yield from automated image analysis was found to be superior to that from manual screening (P=.0443), particularly when the NRBC count was >15. Seven (64%) of 11 XY fetuses were correctly diagnosed by FISH analysis of automatically detected cells, and all discrepancies were restricted to the lower cell-count range. We believe that automated microscopy and image analysis reduce the screening workload, are more sensitive than manual evaluation, and can be used to detect rare HbF-containing NRBCs in maternal blood.     

Effect of concentration, pH, and preservative on in vitro transcorneal permeation of ibuprofen and flurbiprofen from non-buffered aqueous drops

Influence of drug concentration, pH of aqueous drops and some commonly used preservatives on in vitro transcorneal permeation of ibuprofen and flurbiprofen were investigated using goat cornea. Increase in drug concentration in the drops made in normal saline resulted in increase in quantity permeated but decrease in cumulative percent permeation of both drugs. Permeation of each drug from 0.5% drops was maximum at acidic pH (6.4) and decreased with increase in pH of the drops. Normal saline, as a vehicle, favoured permeation of each drug, hence retained in the formulation. Benzalkonium chloride and chlorobutanol enhanced cumulative percent permeation of ibuprofen while benzalkonium chloride and phenyl mercuric nitrate increased permeation of flurbiprofen. Benzalkonium chloride being incompatible with 0.5% drops (pH 6.4) of either drug, chlorobutanol appears suitable for ibuprofen drops and phenyl mercuric nitrate for flurbiprofen drops.     

Contingent monetary reinforcement of smoking reductions, with and without transdermal nicotine, in outpatients with schizophrenia.

This study was conducted to examine the effects of contingent monetary reinforcement (CM) for smoking reduction, with and without transdermal nicotine, on cigarette smoking in individuals with schizophrenia. Fourteen outpatients participated in each of 3 conditions: (a) CM combined with 21 mg transdermal nicotine, (b) CM combined with placebo patch, and (c) noncontingent reinforcement combined with placebo patch. Each condition lasted 5 days. Carbon monoxide levels were measured 3 times daily, and nicotine withdrawal symptoms were measured once daily in each condition. Results indicated that CM reduced smoking but that 21 mg transdermal nicotine did not enhance that effect. These results offer further evidence supporting the efficacy of CM for reducing smoking among people with schizophrenia, but higher doses of nicotine replacement therapy, or another pharmacotherapy, may be needed to enhance that effect. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

An analysis of concentration profiles for fluxes,diffusion depths,and zero-flux planes in multicomponent diffusion

Concentration profiles developed during isothermal, multicomponent diffusion for a single-phase, solid-solid diffusion couple are expressed on the basis of a relative concentration variable for each component and analyzed for the determination of interdiffusion fluxes. The individual concentration profiles intersect at a common cross-over composition where the relative concentrations of all components are identical. New relations are developed for describing internal consistency among the concentration profiles of the various components. A link is made between the cross-over composition and the depths of the diffusion zone on either side of the Matano plane for a diffusion couple. The cross-over composition is interpreted as the average relative concentration of each component over the diffusion zone. The identification of a zero-flux plane from concentration profiles is also described. The analysis offers several advantages in presenting as well as checking the self-consistency of results as illustrated with a single phase Cu-Ni-Zn diffusion couple annealed at 775 °C.     

An in vitro diffusion model for the study of calcification of bovine pericardium tissue

Bovine pericardium (BP) is extensively used for the production of heart valve bioprostheses. BP has excellent mechanical properties but a limited lifespan because of intrinsic subsurface calcification in vivo. In this study, the in vitro mineralization of BP was investigated by a novel diffusion cell model. In two sets of experiments, glutaraldehyde-treated BP membranes were placed between two compartments, both of which contained calcium phosphate solutions made by equilibration of octacalcium phosphate (Exp I) or dicalcium phosphate dihydrate (Exp II) in phosphoric acid. The movement of calcium (Ca), phosphate (P), and protons through the BP membrane was followed throughout the diffusion process. Histology, scanning electron microscopy, wet chemical analysis, and energy dispersive X-ray analyses provided good evidence of subsurface mineralization of BP that resembled in vivo mineral deposition. Energy dispersive x-ray microanalyses found a Ca/P heterogeneity of the early subsurface mineral that formed in the membrane. The use of a diffusion cell to model BP calcification under well-characterized conditions has led to in vitro mineralization that more closely matches that observed in vivo. The results suggest that this in vitro diffusion model can be used to study the mechanism of pathological mineralization. This model has the potential to provide rapid, inexpensive, basic information about the mineralization process.     

Use of dynamic two-dimensional transesophageal echocardiography for renal cell carcinoma with cavoatrial tumor thrombus

Previous investigations involving continuous blood pressure (BP) monitoring have shown an important alteration of the 24-hour BP profile in patients with obstructive sleep apnea syndrome (OSAS). We investigated the impact of REM sleep on the 24-hour BP cycle in 16 severe OSAS male patients (mean respiratory disturbance index = 66 +/- 16 events/hour of sleep), with hypertension (mean BP 162 +/- 21/105 +/- 11 mmHg World Health Organization (WHO) protocol). Two successive nights of polysomnography were performed, and arterial BP was monitored continuously during the second 24-hour period after brachial artery cannulation. During the daytime, subjects were kept awake and supine. At 3 p.m. BP was continuously monitored during quiet supine wakefulness for 20 minutes. Systolic, diastolic and mean BP and heart rate (HR) were analyzed and tabulated in mean values of 5 minute segments. Sleep/wake information were correlated with cardiovascular variables. Each uninterrupted REM sleep period was identified and comparison between the period of quiet supine wakefulness and REM sleep HR and BP values was performed. 8 OSAS patients presented a normal drop of the mean arterial BP during the nocturnal REM sleep periods compared to quiet supine wakefulness (mean value = -10.8 +/- 7.3 mmHg) ("dippers") while the other 8 subjects ("REM sleep non dippers"), revealed an elevated mean arterial BP during REM sleep (mean value = 18.9 +/- 10.9 mm Hg). The absence of the normal circadian BP dip seen during the nocturnal sleep period is considered as an indication of vascular risk. The REM sleep non dipping may play a role in this risk.     

Penetration, permeation, and resorption of 8-methoxypsoralen. Comperative in vitro and in vivo studies after topical application of four standard preparations

The penetration, permeation, and resorption of radioactively labelled 8-Methoxypsoralen was investigated in human skin. Siultaneously, the effects to time and ointment carrier on the penetration kinetics were ascertained. The carriers tested were: vaseline, aqueous wool-wax alcohol ointment, aqueous hydrophilic ointment and polyethylene glycol ointment. The absolute concentrations of 8-Methoxypsoralen were estimated in the horny layer, epidermis and dermis. With the most advantageous carrier, aqueous wool-wax alcohol ointment, 4-6X10(-5) M and 10(-5) M were attained in the epidermis and dermis, respectively. Moreover, it was shown that the substance penetrates rapidly (10 min) into the epidermis and dermis and the high concentrations reached constant over a period of 16 h. Only with a formulation of aqueous wool-wax alcohols is any accumulation at all achieved in the deeper areas of the horny layer. A uniform decrease in drug concentration with increasing depth of the horny layer is found with the other 3 vehicles, whereby slight variations in concentrations pertain from carrier to carrier. 4 h after local application, 8-Methoxypsoralen can be detected in the urine. Regardless of the ointment base employed, 8-Methoxypsoralen is no longer detectable in the urine 40 h after application. In comparison to the oral therapy, the same magnitude of percutaneous resorption into the central compartment is to be derived from the data, if half the body surface is treated locally.     

Novel ventricular apical cannula: in vitro evaluation using transparent, compliant ventricular casts

The geometric configuration of the cannula connection to the left ventricular (LV) apex was studied with respect to several characteristics defining functionality and compatibility. The authors had previously determined, through in vivo studies in sheep, that the design of the cannula used with a dynamic blood pump for LV circulatory support can significantly affect the hemodynamics by improving both the bypass flow rate and the fluid dynamics within the ventricle. The tip of the cannula can aid in preventing wall to wall ventricular collapse, as well as septal shift, due to reduced LV pressure. Proper surgical placement of the cannula with respect to the endocardial surface of the LV can also be simplified by the tip geometry. To investigate the anatomic interaction and fluid dynamics of apical cannulation, transparent compliant casts of bovine LVs were fabricated for in vitro flow visualization. Two different heart geometries were cast, end systolic and end diastolic. The latter was fitted with a pericardial mitral valve and pressurized in a pulsatile fashion to simulate the wall movement of a beating heart. The internal flow and anatomy were visualized with fluorescent particle tracking velocimetry. These studies were performed with conventional cannula tips, as well as a novel, trumpet mouth cannula. The visualization clearly shows the dramatic differences in flow between the geometries tested, and strongly advocates a trumpet mouth design. This novel tip demonstrated excellent placement, beneficial stenting, and improved blood flow by reducing apical stasis and recirculation. Ongoing evaluation of these and future geometries include the application of in vitro endoscopy, quantitative velocimetry, and extension to dilated human ventricles.     

Mutational evidence for control of cell adhesion through integrin diffusion/clustering, independent of ligand binding

Previous studies have shown that integrin alpha chain tails make strong positive contributions to integrin-mediated cell adhesion. We now show here that integrin alpha4 tail deletion markedly impairs static cell adhesion by a mechanism that does not involve altered binding of soluble vascular cell adhesion molecule 1 ligand. Instead, truncation of the alpha4 cytoplasmic domain caused a severe deficiency in integrin accumulation into cell surface clusters, as induced by ligand and/ or antibodies. Furthermore, alpha4 tail deletion also significantly decreased the membrane diffusivity of alpha4beta1, as determined by a single particle tracking technique. Notably, low doses of cytochalasin D partially restored the deficiency in cell adhesion seen upon alpha4 tail deletion. Together, these results suggest that alpha4 tail deletion exposes the beta1 cytoplasmic domain, leading to cytoskeletal associations that apparently restrict integrin lateral diffusion and accumulation into clusters, thus causing reduced static cell adhesion. Our demonstration of integrin adhesive activity regulated through receptor diffusion/clustering (rather than through altered ligand binding affinity) may be highly relevant towards the understanding of inside-out signaling mechanisms for beta1 integrins.     

Flavonoids, dietary-derived inhibitors of cell proliferation and in vitro angiogenesis

Consumption of a plant-based diet can prevent the development and progression of chronic diseases associated with extensive neovascularization, including solid malignant tumors. In previous studies, we have shown that the plant-derived isoflavonoid genistein is a potent inhibitor of cell proliferation and in vitro angiogenesis. In the present study, we report that certain structurally related flavonoids are more potent inhibitors than genistein. Indeed, 3-hydroxyflavone, 3',4'-dihydroxyflavone, 2',3'-dihydroxyflavone, fisetin, apigenin, and luteolin inhibited the proliferation of normal and tumor cells, as well as in vitro angiogenesis, at half-maximal concentrations in the low micromolar range. We have previously demonstrated that genistein concentrations in the urine of subjects consuming a plant-based diet is 30-fold higher than in subjects consuming a traditional Western diet. The wider distribution and the more abundant presence of flavonoids in the plant kingdom, together with the present results, suggest that flavonoids may contribute to the preventive effect of a plant-based diet on chronic diseases, including solid tumors.