Isolation and subfractionation of human peripheral blood mononuclear cells (PBMC) by density gradient centrifugation on Percoll

The use of Percoll for isolation and subfractionation of PBMC and T-lymphocytes by discontinuous and continuous density gradient centrifugation is described: PBMC were isolated from human peripheral blood by discontinuous density gradient centrifugation on Percoll. The use of Percoll instead of Ficoll-Isopaque has the advantage that Percoll, in contrast to Ficoll-Isopaque, does not alter the density of monocytes. Therefore, a better separation of lymphocytes and monocytes was achieved after subsequent continuous density gradient centrifugation on Percoll. E-RFC were isolated by discontinuous density gradient centrifugation after a first low speed centrifugation step banding lymphocytes and SRBC on a Percoll-Ficoll cushion, and a subsequent high speed centrifugation step separating high density rosettes and SRBC from low density non-E-RFC. The advantage of this procedure is the short time of performance and that there is no need to resuspend the lymphocyte/SRBC pellet. PBMC, nph.PBMC T-lymphocytes were further subfractionated by continuous density gradient centrifugation on Percoll. The method described here resulted in a good separation of lymphocytes and monocytes. However, to obtain lymphocyte fractions with minute numbers of contaminating monocytes, a depletion of monocytes prior to further subfractionation of the lymphocytes by continuous density gradient centrifugation is recommended. A marker analysis of T-lymphocytes subfractionated by continuous density gradient centrifugation on Percoll shows that high density T-lymphocytes are enriched in ANAE positive lymphocytes of type 1 and depleted of ANAE positive lymphocytes of type 2. Low density T-lymphocytes are enriched in ANAE type 2 cells and depleted of ANAE type 1 cells. On the other hand, no considerable differences were found when analyzing the T-cells from different fractions for differentiation antigens by means of monoclonal antibodies (anti Lyt 3, OKT4, and OKT8). The results may indicate that subfractionation of T-lymphocytes by continuous density gradient centrifugation on Percoll provided T-cells in different functional states rather than T-cells of distinct subclasses.     

SKY椎体后凸成形术与PVP对骨质疏松多椎体压缩骨折责任椎体选择性治疗的比较

目的探讨SKY椎体后凸成形术与PVP对骨质疏松多椎体压缩骨折责任椎体选择性治疗的疗效对比,评价治疗骨质疏松多椎体压缩骨折责任椎体的有效方法。方法分别于术前、术后2周、术后6月、18个月进行疼痛强度视觉类比评分,观察患者的疼痛症状及生活治疗改善程度。结果 52例手术均成功完成,每个椎体的骨水泥灌注量为3-6mL。两组手术方法术前、术后VAS评分P值分别<0.05,余比较不具统计学意义,两组手术方法术前、术后SF-36评分P值分别<0.05,余比较不具统计学意义。结论无论是SKY椎体后凸成形术或PVP对骨质疏松多椎体压缩骨折选择性治疗均可取得良好的效果,相比PVP,SKY椎体后凸成形术更具优势,但价格昂贵,限制了它的使用。     

Evaluation of Tableting and Tablet Properties of Kollidon SR: The Influence of Moisture and Mixtures with Theophylline Monohydrate

The aim of the study was firstly to investigate the influence of moisture on the tableting and tablet properties of Kollidon SR and secondly to investigate the influence of theophylline monohydrate on the tableting behavior and tablet properties produced from binary mixtures with Kollidon SR. In comparison to Kollidon SR, microcrystalline cellulose (MCC) was used. The glass transition temperature (T_g) of the powder over the whole range of RH (0–90%), and in addition, the T_g of tablets of Kollidon SR were measured. Densities and flowability of the powders were analyzed. The tablets were produced at five different maximum relative densities (ρ_{rel, max}) on an instrumented eccentric tableting machine. They were produced at three different relative humidities (RH), 30%, 45%, and 60% RH for the pure substances and binary mixtures with different ratios of drug and excipient were tableted at 45% RH. The tableting properties were analyzed by 3D modeling, force-displacement profiles, and compactibility plots. First, the T_g of the powder decreased with increasing RH and the T_g of the tablet was 4–8 K lower than the powder. The predominant deformation of Kollidon SR is plastic deformation and Kollidon SR showed a higher compactibility than MCC. The parameters of the 3D model showed an extreme change between 45 and 60% RH, and at higher RH more and more particles deformed elastically. This was confirmed by analysis of force-displacement profiles. At 60% RH, the radial tensile strength of the Kollidon SR tablets was half of the radial tensile strength at 45% RH. The reason is a higher relative energy of plastic deformation than for MCC. This results in a better utilization of the energy to deform the powder into a tablet and the exceeding of the glass transition temperature at higher RH. In conclusion, at 60% RH at the same ρ_{rel, max}, tableting and tablet properties of Kollidon SR are extremely changed since plasticity is significantly higher. In the second part of the study, the insufficient flowability of theophylline monohydrate can be compensated by using Kollidon SR in a mixture with up to 20% theophylline. Further, pressure plasticity ε of MCC and Kollidon SR was lowered in the mixture with theophylline monohydrate. The same is valid for the compactibility. The influence of theophylline monohydrate on the pressure plasticity e of the mixtures was better compensated in the mixture with MCC than in a mixture with Kollidon SR. This compensation was also visible by analyzing the force-displacement-profiles. However, hardly any influence on the radial tensile strength could be detected. Kollidon SR and Kollidon SR mixtures exhibited a higher compactibility than MCC and MCC mixtures. The differences became smaller with increasing theophylline content.     

聚维酮在阿霉素诱导膀胱癌EJ细胞凋亡模型中的作用机制

目的:探讨聚维酮(PVP)在体外加阿霉素(ADM)后诱导EJ细胞发生凋亡的情况以及一些相关机制。方法:应用流式细胞仪(FCM)检测ADM诱导EJ细胞的凋亡率、PVP EJ细胞的凋亡率以及PVP ADM对EJ细胞的诱导凋亡率的情况。结果:PVP对ADM诱导膀胱癌EJ细胞凋亡无影响。结论:PVP在整个诱导过程中并没有协同作用,只起到粘附作用,使得药物停留在EJ细胞表面的作用时间延长,从而达到大量杀伤EJ细胞的作用。     

高黏度骨水泥在骨质疏松性胸腰椎体压缩骨折患者PVP手术中的效果

目的探讨高黏度骨水泥在骨质疏松性胸腰椎体压缩骨折患者PVP手术中的效果。方法回顾性分析2011~2012年本院收治的367例骨质疏松性胸腰椎体压缩骨折患者的临床资料,依据治疗的方法分为高黏度骨水泥椎体成形系统治疗组179例和传统骨水泥椎体成形系统治疗组188例,对比观察两组患者手术前后视觉模拟疼痛评分(Visual Analogue scale,VAS)、压缩椎体前缘高度、Cobb角、骨水泥渗漏情况及术后并发症。结果术后随访1~6个月,两组患者术后VAS评分、压缩椎体前缘高度、cobb角与术前相比均有显著差异(P<0.05),2组患者术后VAS评分无显著差异(P>0.05),术后高黏度骨水泥组患者压缩椎体前缘高度、cobb角与传统骨水泥组患者相比有显著差异(P<0.05);高黏度骨水泥组患者术后并发症发病率与传统骨水泥治疗组比较差异有统计学意(P<0.05)。结论高黏度骨水泥及其椎体成形系统治疗骨质疏松性胸腰椎体压缩骨折具有瞬间高黏度、骨水泥渗漏风险小、手术安全性高、操作简便、预后良好,值得在临床推广应用。     

Identification of five pyrrolidinyl substituted cathinones and the collision‐induced dissociation of electrospray‐generated pyrrolidinyl substituted cathinones

This article reports on the analytical properties of five pyrrolidinyl substituted cathinones: α ‐pyrrolidinononaphenone (α ‐PNP, 1 ), 4‐chloro‐α ‐pyrrolidinopropiophenone (4‐Cl‐α ‐PPP, 2 ), 4‐chloro‐α ‐pyrrolidinovalerophenone (4‐Cl‐α ‐PVP, 3 ), 5‐dihydrobenzofuranpyrovalerone (5‐DBFPV, 4 ), and 2‐(pyrrolidin‐1‐yl)‐1‐(5,6,7,8‐tetrahydronaphthalen‐2‐yl)hexan‐1‐one (β ‐THNPH, 5 ). These identifications were based on liquid chromatography–quadrupole time‐of‐flight‐mass spectrometry (LC–QTOF–MS), gas chromatography–mass spectrometry (GC–MS) and nuclear magnetic resonance spectroscopy (NMR). To our knowledge, no analytical data about α ‐PNP, 4‐Cl‐α ‐PPP, 4‐Cl‐α ‐PVP, and β ‐THNPH have appeared until now, making this the first report on these compounds. Moreover, in order to study the collision‐induced dissociation (CID) characteristic fragmentation routes of pyrrolidinyl substituted cathinones, a total number of 13 pyrrolidinyl substituted cathinones were selected and discussed. The major fragmentation pathways under CID mode are produced, leading to the formation of characteristic ions. Product ions of [M‐C₄H₉N]⁺ and C_nH_2nN⁺ indicate the presence of pyrrolidinyl substitution. Characteristic fragments are also produced via the cleavages of the CH–N(CH₂)₄ bond and the CO‐CHN bond. Copyright © 2016 John Wiley & Sons, Ltd.     

单侧与双侧椎弓根入路椎体成形术治疗高龄骨质疏松性椎体压缩骨折的临床疗效比较

目的探讨单侧与双侧椎弓根入路经皮穿刺椎体成形术治疗骨质疏松性椎体压缩性骨折的临床疗效及价值。方法回顾分析2007年12月～2011年2月我院56例获得6个月以上随访的女性骨质疏松性单个椎体压缩性骨折的临床资料,其中采用单侧椎弓根t入路经皮穿刺椎体成形术治疗30例,双侧椎弓根入路经皮穿刺椎体成形术治疗26例,比较2组患者手术时间、骨水泥填充量、X线照射次数及术后VAS评分。结果单侧组手术时间（25±6）min显著少于双侧组（45±5）rain（t=-13．426,P=0．011）。单侧组骨水泥渗漏率10．0％（3／30）,与双侧组3．8％（1／26）无显著差异（X^2=0．138,P=0．710）。单侧组术中X线曝光（10．5±2．5）次,显著少于双侧组（19．4±3．0）次（t=-12．110,P=0．000）。2组术后24h、术后6、12个月VAS评分无统计学差异（P〉0．05）。结论单侧与双侧椎弓根入路经皮穿刺椎体成形术治疗骨质疏松性椎体压缩性骨折均能取得良好效果。单侧穿刺方法手术时间短,X线暴露次数少;双侧入路穿刺方法手术操作相对简单。     

稳定的阿戈美拉汀晶型Ⅰ的制备方法

开发了一种稳定的阿戈美拉汀晶型I的制备方法。该方法采用在无水乙醇-正庚烷溶剂体系下添加PVP或在无水乙醇-水体系下添加HPMC的抗溶剂结晶法来实现。通过不同溶剂体系下高分子材料种类的筛选、不同浓度高分子材料的影响及溶剂体系中良溶剂和抗溶剂比例的影响的研究,确定了制备阿戈美拉汀晶型I的工艺。通过加速稳定性实验和长期稳定性实验对比表明,采用加高分子材料制备方法得到的阿戈美拉汀晶型I,具有较好的稳定性。     

Design and Evaluation of Bilayer Pump Tablet of Flurbiprofen Solid Dispersion for Zero-Order Controlled Delivery

In this study, a bilayer osmotic pump tablet of flurbiprofen (FP) solid dispersions (SDs) was developed to increase the solubility of the poorly soluble drug and controlled drug release at a constant rate. Based on the investigation of thermodynamic properties the drug, the carrier, and the calculation of the solubility parameters, the FP-SD was prepared by hot-melt extrusion technique with the povidone (PVP) VA64 carrier. Then, central composite design-response surface methodology was used to evaluate the influence of factors on the responses. Consequently, PVP VA64 was selected as the carrier for preparing FP-SD. The results of differential scanning calorimetry and X-ray confirmed that FP in FP-SD was in an amorphous state. FTIR indicated that the intermolecular hydrogen bond probably formed between FP and PVP VA64 in FP-SD. Correlation of release profiles to zero-order kinetics was significant (R² = 0.9939). The mathematical models had good predictability because the deviation was less than 1% between the predicted value and measured value. These results demonstrated that FP-SD osmotic pump tablets successfully increased the solubility of FP and controlled the release of FP at a constant rate.     

丘脑底核电刺激治疗苍白球毁损术后的帕金森病(12例报告)

目的探讨帕金森病(Parkinson's disease,PD)苍白球腹后部毁损术(posteroventral pallidotomy,PVP)后再行丘脑底核(subthalamic nucleus,STN)脑深部电刺激术(deep brain stimulation,DBS)的可行性、术中电生理学特点和治疗结果。方法应用MR和微电极记录技术进行靶点定位,对12例单侧PVP术后症状再次加重的PD患者实施STN-DBS手术,其中4例行毁损灶对侧的STN-DBS,8例行双侧STN-DBS。结果STN-DBS对本组12例PD患者症状有不同程度的改善,双侧STN-DBS的效果尤为明显,术后3个月的UPDRS运动及ADL评分较术前明显减少(P<0．05或0．01),美多巴的用量明显减少(P<0．01),无明显术后并发症。术中电生理记录显示毁损灶同侧的细胞放电明显低于正常情况。结论曾行单侧PVP的PD患者如面临二次手术,可以选择DBS手术,以双侧STN的DBS效果最好,可减少药物用量。