Preparation and evaluation of lectin-conjugated PLGA nanoparticles for oral delivery of thymopentin.

The purpose of this study was to design and evaluate lectin-conjugated PLGA nanoparticles for oral delivery of thymopentin. Thymopentin loaded PLGA nanoparticles (TP5-NPs) were prepared by a double emulsion-solvent evaporation technique. Novel WGA-PLGA conjugates were synthesized by coupling the amino groups of wheat germ agglutinin (WGA) to the carbodiimide-activated carboxylic groups of PLGA, and were incorporated into nanoparticles preparation to take mucoadhesive properties. Important characteristics such as particle size, zeta potential, entrapment efficiency, storage stability, as well as in vitro drug release behavior were investigated. The retention of biorecognitive activity of WGA after covalent coupling was confirmed by haemagglutination test. In vitro experiments with pig mucin (PM) demonstrated that the conjugation of WGA enhanced the interaction about 1.8-4.2 fold compared with that of the non-conjugated nanoparticles, and still exhibited sugar specificity. The pharmacodynamical studies on oral administration of WGA-TP5-NPs were performed in FACScan flow cytometry. The values of CD4(+)/CD8(+) ratios were significantly increased compared with that of TP5-NPs (p<0.01). The enhanced uptake was related to the increasing of WGA content on nanoparticles. These results confirmed that the conjugation of WGA onto PLGA nanoparticles effectively improved the intestinal absorption of TP5 due to specific bioadhesion on GI cell membrane.     

A型肉毒毒素多表位串联体的设计、表达、纯化及鉴定

目的:设计、表达两个A型肉毒毒素多表位串联体.方法:从文献报道的A型肉毒毒素(botulinum neurotoxin type A,BoNT/A)的表位及生物信息学方法预测得到的B细胞表位中遴选表位,并加入适当的辅助性元件,设计多表位串联体A、B.对其基因进行优化后,经重叠PCR方法合成串联体A、B的全长基因.分别插入原核表达载体pQE-30,再转化E.coli M15[pREP4]感受态细胞中进行诱导表达,以金属螯合亲和层析法纯化重组蛋白,并进行鉴定.结果与结论:成功设计并构建了两个多表位串联体A、B,并在E.coli中以包涵体形式获得高效表达.Ni-NTA法纯化后分别获得纯度大于92.2%、99%的重组串联体A、B蛋白,并经透析复性法复性.Western印迹和间接ELISA显示纯化、复性后的重组蛋白与抗天然BoNT/A马血清有特异性结合反应.     

分步骤法维护三向瓣膜式PICC导管留置效果观察

目的预防外周中心静脉导管置管(PICC)后滑脱及体外部分非人为移入体内,减少局部炎症反应,防止导管性静脉炎发生.方法将42例接受PICC的肿瘤化疗患者随机分为观察组(n=22)和对照组(n=20),观察组采用分步骤法维护,即分步骤地更换透明贴膜及肝素帽(或先换肝素帽,再换透明贴膜),并将导管体外部分直形固定;对照组采用传统法维护,即同时更换透明贴膜及肝素帽,导管体外部分S形固定.结果观察组静脉炎,穿刺点渗液、硬结,导管移位发生率显著低于对照组(P<0.05,P<0.01).结论行PICC导管维护时,分步骤更换透明贴膜及肝素帽,将导管体外部分直形固定,可有效防止导管滑脱及体外部分进出体内,减少局部炎症反应,预防导管性静脉炎.     

支气管结核在纤维支气管镜下治疗方法的探讨

目的 :探讨支气管结核在纤维支气管镜下治疗的方法及价值。方法 :对 2 5例支气管结核病人 ,在全身抗结核治疗的同时分别实施镜下注药治疗、微波接触式辐射治疗和球囊扩张术 局部注药治疗。结果 :在全身抗结核治疗的同时运用上述几种镜下治疗方法 ,支气管局部病灶较镜下治疗前均有明显好转。结论 :利用纤维支气管镜对支气管结核病人进行局部治疗 ,可加快病灶的吸收和症状的改善 ,值得推广应用     

静脉输液加药后的微粒变化

本文对我院急诊病人常用的供静脉给药的22种药物、42组配伍情况,制成静脉加药输液,在超净工作台条件下,用KF—4型微粒计数器,测定了168次微粒数,结果显示输液加药后微粒虽有增加,但均在药典规定范围内。输液中加1种、2种、3种药物后所产生的微粒数分别为x_1=5.52(n_1=16);x_2=8.17(n_2=18)、x_3=10.25(n_3=8),经统计处理,三组间无显著差异(P<0.05)。本文对产生微粒的药物、注射器、操作环境等因素进行了分析。     

癌痛从痰论治探讨

癌痛是晚期癌症患者的主要症状之一,属于中医痛证范畴。痰是产生癌痛的重要中医病机,消痰法是治疗癌痛的重要方法。     

血管紧张素对增生性瘢痕成纤维细胞纤维连接蛋白合成的影响

目的：探讨血管紧张素Ⅱ（AngⅡ）及其Ⅰ型、Ⅱ型受体阻断剂和钙调神经磷酸激酶（CaN）的阻滞利环胞素A（CsA）对增生性瘢痕来源的成纤维细胞纤维连接蛋白mRNA和蛋白表达的作用。方法：体外分离培养增生性瘢痕成纤维细胞，分别将一定浓度的AngⅡ（10^-9～10^-5mmol／L），和AngⅡ10^-6mmol／L加上不同阻滞剂losartan、PD123319、环胞素A（浓度均为10^-5mmol／L）加入细胞培养液中刺激48h，分别采用逆转录聚合酶链反应（RT—PCR）及Westernblot印迹方法检测增生性瘢痕成纤维细胞纤维连接蛋白（FN）的mRNA及蛋白表达。结果：体外成功地培养增生性瘢痕成纤维细胞。经AngⅡ刺激48h后，FN mRNA和蛋白表达量明显增高，增高程度与AngⅡ浓度呈正比；加入losaartan和环胞素A后，FN原mRNA和蛋白表达量较单用AngⅡ组下降，而加入PD123319后，FNmRNA和蛋白表达量较单用AngⅡ组无明显改变。结论：AngⅡ可促增生性瘢痕成纤维细胞的FN合成，可能在增生性瘢痕的发展过程中起重要作用，该作用主要通过AngⅡ的Ⅰ型受体介导完成，该作用可能与CaN信号通路有关。     

提壶揭盖法治疗前列腺增生症82例

笔者自1995年以来，宗提壶揭盖法，用补中益气汤加减治疗前列腺增生82例疗效满意，报告如下。     

不同浓度5-溴脱氧尿嘧啶核苷标记山羊骨髓基质干细胞的体外研究

Objective To explore the feasibility of labeling and tracing in vitro goat bone marrow mesenchymal stem cells (BMSCs) by bromodeoxyuridine (BrdU) on the basis of investigation of its optimal concentration, incubating time and cytotoxicity. Methods A healthy goat, aged 10 months old, male, weighing 32 kg, was used in this study. Bone marrow was aspirated. BMSCs were isolated and cultured using the adherence method in vitro. The fourth passage of BMSCs (P4) were incubated with BrdU at 5, 10, 15, 20 μmol/L as 5, 10, 15, 20 μmol/L BMSC groups. Cells were not labeled by BrdU as negative control. The following parameters were measured: induction, differentiation and determination of goat BMSCs; the optimal mass concentration and incubation time of 5-BrdU labeling; cell positive rate at 12, 24, 48 and 72 hours in each group using immunofluoreseenee; the cell survival rate after various concentrations of BrdU ladling by trypan-blue exclusion. Results The morphology of the primary and passage goat BMSCs was fusiform in shape. Goat BMSCs could differentiate into osteoblasts and chondrocytes following induction. BMSC nucleus showed green fluorescence under fluorescence microscope after being labeled by BrdU. The mean labeling rate increased with the increase in the concentration and incubation time of BrdU, and reached to (93.32± 3.25)% after incubation in 15 μmol/L, BrdU for 48 hours. There were no significant differences between 15 μmol/L BrdU for 72 hours, 20 μmol/L BrdU for 48 hours and 72 hours (P > 0.05), or between the other groups or time points (P < 0.05). The labeling rate of the blank control group was 0. The cell survival rate was all above 90% (P > 0.05). Conclusions BrdU can be used as a labeling marker for goat BMSCs. When the concentration is 15 μmol/L and the incubation time is 48 hours, the optimal labeling effect can be achieved. Goat BMSCs labeled with BrdU is of high efficiency and safety.     

慢性肾脏病非透析患者脑钠素与动脉粥样硬化及心功能的关系

目的 研究脑钠素（BNP）与慢性肾脏病（CKD）非透析患者动脉粥样硬化及心功能不全的关系。 方法 采用双抗夹心免疫荧光法检测203例CKD非透析患者与16例高血压患者对照组全血BNP水平，分析其与颈动脉超声结果、心脏彩超结果及既往心血管疾病史的关系。 结果 CKD非透析患者BNP水平与对照组相比显著升高[M（范围）：54.40(15.10～ 173.00) ng/L比9.35(7.35～15.00) ng/L，P < 0.01]。Spearman相关分析显示CKD患者BNP与颈动脉内膜中层厚度（IMT）、左室心肌重量指数（LVMI）等呈正相关。存在颈动脉斑块、左室肥厚或既往发生过心血管事件的患者血BNP水平显著增高。多元回归分析显示LVMI、既往心血管事件均是影响BNP水平的独立因素。 结论 CKD非透析患者BNP水平和动脉粥样硬化性疾病、左室肥厚及心功能不全相关，提示BNP水平可作为一项评价CKD非透析患者心功能及动脉粥样硬化的敏感生物学指标。