神经症的性压抑与心理因素的相关性研究

研究神经症患者潜在人格与性压抑和情绪的相关性.方法:对124例神经症患者进行了婚姻质量问卷(性生活、婚姻质量部份)、抑郁自评量表(以下简称SDS)、焦虑自评量表(以下简称SAS)及自编的性生活调查问卷、主题统觉测验(以下简称TAT)测评.结果:1、神经症伴性障碍组TAT潜在人格与对照组相比,在支配、性、援助、躯体社会攻击、财产破坏欲求、自我攻击、情绪变化欲求和情绪言语攻击性、躯体社会攻击、引诱、缺乏、丧失、灾害、丧失支持、躯体伤害压力上均较对照组显著增高,而成就欲求下降(P<005-001);与神经症无性障碍组相比,神经症伴性障碍组的成就、支配、教养、性欲求得分显著增高,情绪言语攻击欲求得分下降(P<001～005);2、神经症伴性障碍组的性生活质量低,性频度、性后快感均低.3、SDS、SAS与TAT中的负性欲求呈明显的相关性,性行为调查与婚姻质量量表中的性行为呈正相关.结论:神经症的性压抑与其潜在人格有密切的相关性.     

31P solid-state NMR based monitoring of permeation of cell penetrating peptides into skin

The main objective of the current study was to investigate penetration of cell penetrating peptides (CPPs: TAT, R₈, R₁₁, and YKA) through skin intercellular lipids using ³¹P magic angle spinning (MAS) solid-state NMR. In vitro skin permeation studies were performed on rat skin, and sections (0–60, 61–120, and 121–180 μm) were collected and analyzed for ³¹P NMR signal. The concentration-dependent shift of 0, 25, 50, 100, and 200 mg/ml of TAT on skin layers, diffusion of TAT, R₈, R₁₁, and YKA in the skin and time dependent permeation of R₁₁ was measured on various skin sections using ³¹P solid-state NMR. Further, CPPs and CPP-tagged fluorescent dye encapsulate liposomes (FLip) in skin layers were tagged using confocal microscopy. The change in ³¹P NMR chemical shift was found to depend monotonically on the amount of CPP applied on skin, with saturation behavior above 100 mg/ml CPP concentration. R₁₁ and TAT caused more shift in solid-state NMR peaks compared to other peptides. Furthermore, NMR spectra showed R₁₁ penetration up to 180 μm within 30 min. The results of the solid-state NMR study were in agreement with confocal microscopy studies. Thus, ³¹P solid-state NMR can be used to track CPP penetration into different skin layers.     

Tamoxifen induces resistance to activated protein C

Introduction

The estrogen antagonist tamoxifen (TAM) increases the thrombotic risk similar to estrogen containing oral contraceptives (OC). In OC users this risk is attributed to alterations of hemostasis resulting in acquired resistance to activated protein C (APC). TAM-induced APC resistance has not been reported yet.

Materials and Methods

Blood samples were collected prospectively from women with breast cancer before (n = 25) and monthly after start of adjuvant TAM treatment (n = 75). APC resistance was evaluated on basis of the effect of APC on the endogenous thrombin generation potential. To detect increased in vivo APC generation APC plasma levels were measured using a highly sensitive oligonucleotide-based enzyme capture assay. Routine hemostasis parameters were measured additionally.

Results

APC sensitivity decreased by 41% (p = 0.001) compared to baseline after one month of TAM application and remained significantly decreased during the study period. Free protein S increased (p = 0.008) while other analyzed procoagulant factors, inhibitors, and activation markers of coagulation decreased or did not change significantly. In five patients the APC concentration increased to non-physiological levels but an overall significant increase of APC was not observed.

Conclusions

This is the first study showing acquired APC resistance under TAM therapy. Acquired APC resistance might explain the increased thrombotic risk during TAM treatment. Observed changes of hemostasis parameters suggest different determinants of TAM-induced APC resistance than in OC-induced APC resistance. The presence of acquired APC resistance in TAM patients warrants further evaluation if these patients may benefit from antithrombotic prophylaxis in the presence of additional thrombotic risk factors.     

穿膜肽TAT修饰载丹酚酸B脂质体的制备及其抑制人皮肤成纤维细胞增殖与迁移初步研究

目的制备具有防治增生性瘢痕(HS)作用的载丹酚酸B的穿膜肽TAT修饰脂质体(SAB-TAT-LIP),建立其质量评价方法,并初步考察其对体外人皮肤成纤维(HSF)细胞增殖和迁移的影响。方法采用pH梯度逆向蒸发法制备脂质体,超滤法测其包封率,以包封率为评价指标,采用Box-Behnken响应面法优化脂质体的处方工艺;考察其形态、粒径、Zeta电位、体外释放、体外透皮吸收和稳定性等理化性质;在此基础上采用MTT法考察其对HSF细胞增殖的作用,采用划痕法和Transwell小室法考察其对HSF细胞迁移和侵袭的作用。结果 SAB-TAT-LIP的药物包封率为(86.70±0.85)%,平均粒径为(219.90±5.09)nm,Zeta电位(-9.25±0.92)m V,体外24h累积释放率为62.49%,无突释效应,体外32h皮肤累积透过率为17.21%,透过速率为(28.33±4.9)μg/(cm2·h),真皮层滞留量为(44.39±6.87)μg/cm2,4℃放置10d稳定性良好。SAB-TAT-LIP能够显著地抑制HSF细胞的增殖、迁移和侵袭,与对照组相比,差异显著(P0.01)。结论优化得到的SAB-TAT-LIP包封率较高、粒径较小,体外释放和透皮行为均满足局部透皮给药制剂的体外释放和透皮规律,对体外HSF细胞的增殖、迁移和侵袭具有抑制作用。     

Conjugates of TAT and folate with DOX-loaded chitosan micelles offer effective intracellular delivery ability

The key for better antitumor efficacy is to improve the specificity of antitumor drugs for tumor cells and diminish their cytotoxicity to normal tissues. Targeted nanoparticles as antitumor drug delivery system can resolve this problem. In this study, we prepared folate and TAT (arginine-rich cell-penetrating peptide) modified N-PEG-N′-octyl-chitosan to form the folate/TAT-PEG-OC micelles. Then, the molecular structure, morphology, size distribution and bio-safety of the micelles were characterized. In order to investigate the drug-loading capacity of folate/TAT-PEG-OC micelles, doxorubicin (DOX) was used as model drug to prepare DOX-loaded chitosan micelles. Here, the confocal microscopy was used to evaluate the cellular uptake of DOX/folate/TAT-PEG-OC micelles, while the self-built NIR imaging system was used to evaluate the dynamic behavior of ICG-Der-01/folate/TAT-PEG-OC micelles in vivo. Our results demonstrate that the dual-modified PEG-OC micelles not only have good morphology, uniform size distribution and excellent drug loading capacity, but also show a strong capability for the efficient intracellular uptake and enhanced targeting behaviors in a folate receptor positive tumor model (Bel-7402 human hepatocellular cells). All these results suggest the potential application of folate/TAT-PEG-OC micelles in the targeted diagnosis and therapy to different kinds of folate receptor positive tumors.     

Characterization of two novel mutations of the antithrombin gene observed in Japanese thrombophilic patients

We investigated the molecular basis of reduced functional levels of antithrombin (AT) in two individuals suffering from thromboembolic events. In each case direct sequencing of amplified DNA revealed 13,260-13,262 del in one patient and 2511C>A in the other patient, predicting a heterozygous E381del and P16H, respectively. Both patients had no 20210A allele and factor V Leiden mutation. To understand the molecular mechanism responsible for antithrombin deficiency, stable expression experiments were performed using HEK293 cells transfected with the expression vector containing the wild-type or the mutated recombinant cDNA. In these experiments, the media levels of the two mutated antithrombins were the same as that of wild type, but the specific activity of the E381del mutant decreased significantly compared with that of wild type. These results showed that the E381del mutation was responsible for type II deficiency, whereas the other mutation, P16H, did not produce any definite abnormality which could contribute to antithrombin deficiency.     

A pilot investigation of mild hypothermia in neonates receiving extracorporeal membrane oxygenation (ECMO)

OBJECTIVE: To investigate the safety and feasibility of using mild hypothermia in neonates receiving extracorporeal membrane oxygenation (ECMO).Study design A prospective, nonrandomized pilot study of 25 neonates referred for ECMO. Whole body cooling was achieved by adjustment of the temperature of the extracorporeal circuit water bath. Five groups (N=5 per group) were each studied for the first 5 days of ECMO. The first group was maintained at 37 degrees C throughout the study period. Subsequent groups were cooled to 36 degrees C, to 35 degrees C, and, finally, to 34 degrees C, respectively, for 24 hours and the final group to 34 degrees C for 48 hours before being rewarmed to 37 degrees C. Patients were carefully assessed clinically and biologically. In addition to routine laboratory tests, cytokines (IL-6 and IL-8), complement (C3a), and molecular markers of coagulation (thrombin/antithrombin III [TAT], antithrombin III, and plasmin-alpha2plasminogen) were measured. RESULTS: No major clinical or circuit problems were noted during cooling or rewarming. In particular, there were no problems of bleeding or cardiac arrhythmia. No significant difference was found between groups in terms of molecular markers of coagulation, complement, cytokines, and platelet transfusions. CONCLUSIONS: Applying mild hypothermia (34 degrees C) for 24 or 48 hours to neonates receiving ECMO is both feasible and safe.     

The mechanisms of cell death in focal cerebral ischemia highlight neuroprotective perspectives by anti-caspase therapy

A number of studies have validated the importance of caspase activation in ischemia-induced brain damage. Caspases participate in both the initiation and execution phases of apoptosis, and play a central role in neuronal death after global cerebral ischemia. In focal ischemia, apoptosis occurs in the penumbra during the secondary phase of expansion of the lesion. However, ultrastructural and biochemical analysis have also shown signs of apoptosis in the initial lesion, or infarct core, which is traditionally considered necrotic. Specific caspase pathways are activated in the core and in the penumbra, and participate in both cytoplasmic and nuclear apoptotic events, notwithstanding their initial classification as activator or initiator caspases. This confirms previous suggestions that caspase inhibition holds tremendous neuroprotective potential in stroke and other apoptosis-related degenerative diseases. Consequently, two new approaches, aimed at treating stroke-induced brain damage by anti-apoptotic molecules, are being developed in academic and industrial laboratories. These are based, respectively, on the use of small peptide sequences corresponding to the preferred cleavage site of a caspase, and on genomic constructions derived from the fusion of endogenous anti-caspase molecules with a protein transduction domain from the human immunodeficiency virus-1. Fusion proteins containing endogenous caspases inhibitors efficiently counteract apoptosis in vitro. In in vivo models of focal cerebral ischemia, fusion proteins successfully cross the blood brain barrier and protect cells from ischemic death. This new approach by protein therapy could prove to be an interesting alternative for the reduction of the dramatic consequences of stroke, provided that the long-term efficiency of this protection in terms of functional recovery is demonstrated.     

Total testing process: appropriateness in laboratory medicine

BACKGROUND: The need to reduce costs in Laboratory Medicine is often related to the possibility of reducing test requests without taking into account patients' outcomes. Therefore, the term "appropriateness" in Laboratory Medicine as referred to the specific steps (pre-analytical, analytical, post-analytical) and related to the clinical process could allow the improvement of clinical effectiveness and economic efficiency. METHODS: Our experience has shown an improvement in analytical appropriateness (reorganization and re-engineering by Laboratory automation) and pre-analytical appropriateness (critical revision of the panel for cardiac markers) by evaluating the workload and errors rate in the pre-analytical phase. RESULTS: We obtained an economic saving (119,580 euro/year) in cardiac markers request (analytical appropriateness: 60%, pre-analytical appropriateness: 40%) and also an improvement in clinical appropriateness (diagnosis and therapy). CONCLUSIONS: Our data confirm the need to improve communications between physicians and Laboratory Medicine as regards the pre-analytical step and to implement educational programs for defining criteria and procedures. Appropriateness in analytical and post-analytical steps contribute to achieve economic saving (Core lab, POCT) and improvement of the turn-around time (TAT).