白喉和破伤风类毒素的定量测定2.单向辐射状免疫扩散试验和火箭免疫电泳试验与絮状反应试验之比较

作者测定了8种白喉类毒素和9种破伤风类毒素。絮状反应和单向辐射状免疫扩散试验(SRID)所用的白喉和破伤风抗毒素为超免疫马血清,火箭免疫电泳试验则用家兔血清。每种类毒素均作4个稀释度,抗毒素则适量加入1%琼脂糖凝胶中。将试验类毒素与参考类毒素配对,随机加入凝胶板中。结果表明,火箭免疫电泳试验与絮状反应试验相比,所测白喉类毒素絮状反应     

一种快速和敏感的吸附破伤风类毒素的鉴别试验

本文介绍用间接血凝抑制(HAI)试验作为吸附破伤风类毒素(TT)的鉴别试验。WHO推荐的絮状试验或凝胶免疫沉淀试验,都需要在柠檬酸钠溶液中溶解性剂,需2天时间才能得到结果。此外,絮状试验还常见假沉淀带。     

19A型肺炎链球菌多糖结合物中游离多糖含量测定方法的建立

目的 建立19A型肺炎链球菌多糖(Streptococcus pneumoniae type 19A polysaccharide,19APS)结合物中游离PS含量的测定方法.方法 在一定条件下分别用1、3和5 g/L脱氧胆酸钠(sodium deoxycholate,DOC)溶液(pH7.3)沉淀不同浓度的破伤风类毒素(tetanus toxoid,TT),确定不同浓度DOC沉淀载体蛋白TT的浓度范围.以1 g/L DOC分离结合19 APS(19APS-TT)和游离19APS,测定上清液中的游离PS含量,并验证该DOC沉淀法的准确度和精密度.结果 分别以1和3 g/L DOC沉淀19APS-TT时,TT浓度最好分别控制在＜150和＜200μg/ml.该DOC沉淀法的准确度和精密度均良好,加标PS回收率为94.0％～107.2％,相对标准偏差均＜4％.结论 成功建立了19APS-TT中游离PS含量的测定方法.     

检测C群脑膜炎球菌结合疫苗中游离多糖含量的脱氧胆酸钠沉淀法的建立

 目的  建立检测C群脑膜炎球菌多糖-破伤风类毒素结合疫苗（group C  meningococcal polysaccharide -tetanus toxoid conjugate vaccine, CPS-TT）中游离多糖的脱氧胆酸钠沉淀法。 方法  在不同pH条件下用脱氧胆酸钠（sodium deoxycholate,DOC）对CPS和TT进行沉淀,确定可有效分离游离多糖和结合多糖的最适沉淀条件,并对该法进行验证。 结果  DOC沉淀法的最适pH为3.5,在此pH条件下用DOC沉淀法检测3批CPS-TT显示,上清TT含量平均为2.3%。DOC沉淀法具有较好的准确度和精密度,3批CPS-TT的批内变异系数均小于5%。 结论  DOC沉淀法可用于C群脑膜炎球菌结合疫苗的质量控制。     

体外游离或微囊化破伤风类毒素稳定性的理化及免疫学研究

近年来药物控制释放系统的发展受到人们关注,由丙交酯和乙交酯聚合物制备的微球已作为单剂破伤风类毒素菌苗的新的释放系统。本文采用圆二色谱(CD)、荧光光谱、快速蛋白液相层析(FPLC)凝胶过滤层析技术及免疫化学方法,比较在不同条件下游离或微囊化破伤风类毒素在3个月内的结构及抗原完整性的变化。     

脉冲式破伤风类毒素聚乳酸微球动物免疫效果研究

目的　研究破伤风类毒素聚乳酸微球在动物体内的免疫效果及影响因素。方法　观察破伤风类毒素聚乳酸微球免疫动物后1年中,动物血清抗体反应及免疫记忆反应的规律。结果　微球中抗原释放为脉冲式,释放时间与聚乳酸分子量、微球载药量及粒径有关,微球引起的抗体反应显著高于破伤风类毒素溶液一次免疫,与溶液3次免疫效果相似。结论　创制了一种新型脉冲式释药系统,一次注射完成全程免疫,有巨大的应用价值     

对现行的破伤风和白喉类毒素效力试验方法的重新评价

本文在明确指出现行的破伤风和白喉类毒素效力试验方法的问题并作出重新评价后,提出用絮状反应试验及有关物理化学特性检测来替代现行的动物效力试验,以保证类毒素质量的稳定性。     

成人抗白喉再接种 Ⅱ.基础免疫后20年用不同剂量的白喉类毒素与破伤风类毒素联合进行再接种

作者对273名18～25岁的新兵进行白喉类毒素(D)和破伤风类毒素(T)联合制剂(DT)再接种。将受试者随机分成A、B和C3组,分别接种6个絮状反应单位(Lf)T、6LfT+2LfD和6LfT+5LfD。每ml制剂中含1mg Al(OH)_3作为佐剂。接种后1周观察副反应。接种前后4周采集静脉血,用ELISA测定破伤风抗体浓度,用Vero细胞组织培养中和法测定白喉抗体浓度。抗体浓度达0.01IU/ml为保护水平。     

成人接种白喉-破伤风类毒素的反应

过去,成人接种白喉-破伤风类毒素(DT)后的反应率很高,此与制品中的类毒素剂量和接种者先前的免疫状态有关。后来由于技术的进步,此疫苗有很多改进,主要是类毒素的提纯和标准剂量的确定;白喉类毒素剂量由25Lf单位(絮状反应单位)减至2Lf单位,可以生成足够的抗体,但反应率大为降低。近年由于白喉的爆发导致对成人和儿童二者都进行广泛的DF接种。 1975年秋阿拉斯加散在发生12例白喉,流行病学调查未发现其间的传播关系,接触者中发现大批无症状带菌者,因而刘成人采取DT普遍接种。所用DT制剂为明矾沉淀     

用浓缩白百破疫苗两针法进行婴儿免疫

预防白喉、百日咳、破伤风的传统方法是注射白百破混合制剂,共注三针,每次间隔一或两个月。本文报导在菲律宾用浓缩白百破两针法免疫,同时用常规白百破及吸附白喉类毒素作对照,比较免疫效果及反应性。观察分成三组,即A组(174人)注射吸附白类,每人份含15絮状反应单位(Lf)、磷酸铝1.35mg,由马尼拉Alabang研究所制造;B组(177人)注射浓缩白百破,每人份分别含白喉类毒素15Lf,破伤风类毒素10Lf,百日咳10个国际比浊单位(IOU),磷酸铝3mg,由荷兰Rijks研究所生产;C组(171人)注射常规白百破,每人份分别含白喉类毒素15Lf、破伤风类毒素7.5Lf、百日咳10IOU、磷酸铝1.35mg,由马尼拉Alabang研究所生     

双氯芬酸二乙胺凝胶的体外透皮扩散研究

目的:制备双氯芬酸二乙胺凝胶并考察其释药影响因素。方法:采用正交试验,对双氯芬酸二乙胺凝胶进行体外透皮试验,以透皮扩散速率常数为考察指标。结果:双氯芬酸二乙胺凝胶的处方中异丙醇浓度为15%,丙二醇为10%,卡泊姆934为0.5%时,药物的透皮扩散速率最快。结论:卡泊姆934的浓度为影响透皮扩散的主要因素。     

盐酸莫西沙星耳用凝胶的制备及质量控制

目的 制备盐酸莫西沙星耳用凝胶并建立其质量控制方法.方法 以卡波姆-940为基质制备凝胶,采用高效液相色谱法测定盐酸莫西沙星含量,并考察其稳定性.结果 盐酸莫西沙星进样量在0.4μg～3μg范围内与峰面积积分值线性关系良好（r=0.9999）,平均回收率为99.82%（RSD=0.43%,n=9）.该制剂对家兔耳未见刺激性发生.结论 本制剂制备工艺简单,质量控制易行,稳定性好,有良好的临床使用价值.     

Nanogel for dermal application of the triterpenoids isolated from Ganoderma lucidum (GLT) for frostbite treatment

Abstract

Objective: The purpose of this study was to formulate stable Ganoderma lucidum (GLT) nanogels suitable for topical delivery with a view to improve the therapeutic effect for frostbite.

Methods: GLT nanosuspensions were formulated using the high-pressure homogenization technique and then suitably gelled for characterized. In order to confirm the advantages of GLT nanogel for dermal application, skin permeation studies in vitro and pharmacodynamic evaluation in vivo were studied and compared with GLT–carbopol gel.

Results: The particle size analysis and SEM studies revealed that GLT nanosuspensions were still stably kept their particle size after suitably gelled by carbopol preparation. The drug content, pH, and spreadability of the GLT nanogel was found to be 99.23?±?1.8%, 6.07?±?0.1, and 26.42 (g·cm)/s, which were within acceptable limits. In vitro permeation studies through rat skin indicated that the amount of GLT permeated through skin of GLT nanogel after 24?h was higher than GLT–carbopol gel, and GLT nanogel increased the accumulative amount of GLT in epidermis five times than GLT–carbopol gel. No oedema and erythema were observed after administration of GLT nanogel on the rabbits' skin. Pharmacodynamic study showed that GLT nanogel was more effective than GLT–carbopol gel in treatment of frostbite.

Conclusion: The GLT nanogel possess superior therapeutic effect for frostbite compared with the GLT–carbopol gel, which indicates that nanogels are eligible for the use as a suitable nanomedicine for dermal delivery of poorly soluble drugs such as GLT.     

纳米硫凝胶的制备及质量控制

目的制备纳米硫凝胶,并建立其质量控制标准。方法液相法合成纳米硫,以卡波姆940为主要基质制备纳米硫凝胶,采用pH检测、含量测定、稳定性考察等方法控制凝胶质量,并进行皮肤刺激性试验。结果合成的纳米硫粒径为50～80 nm。纳米硫凝胶在室温下均匀细腻,流动性较好,pH为6.5,其含量和稳定性等均符合质量控制标准。结论纳米硫凝胶制备工艺可行,性质稳定,质量可控。     

Evaluation of Carbopol-Methyl Cellulose Based Sustained-Release Ocular Delivery System for Pefloxacin Mesylate Using Rabbit Eye Model

The major purpose of this study was to develop and characterize a series of carbopol- and methyl cellulose–based solutions as the in situ gelling vehicles for ophthalmic drug delivery. The rheological properties, in vitro release as well as in vivo pharmacological response of a combination of polymer solutions, including carbopol and methyl cellulose, were evaluated. It was found that the optimum concentration of carbopol solution for the in situ gel-forming delivery systems was 0.3% (w/w), and that for methyl cellulose solution was 1.5% (w/w). The mixture of 0.3% carbopol and 1.5% methyl cellulose solutions showed a significant enhancement in gel strength in the physiological condition; this gel mixture was also found to be free flowing at pH 4.0 and 25°C. The rheological behaviors of carbopol/methyl cellulose solution were not affected by the incorporation of the drug. Drug levels in the aqueous humor of the rabbits were well above the MIC-values of relevant bacteria after 12 hours, the results of an optimized formulation containing 0.18% of pefloxacin mesylate compared well with the 0.3% marketed eye drop formulation, indicating our formulation to be significantly better considering that a similar effect was obtained at half the concentration. Both the in vitro release and in vivo pharmacological studies indicated that the carbopol/methyl cellulose solution had better ability to retain drug than did the carbopol or methyl cellulose solutions alone. The results demonstrated that the carbopol/methyl cellulose mixture can be used as an in situ gelling vehicle to enhance the ocular bioavailability of pefloxacin mesylate.     

Evaluation of carbopol-methyl cellulose based sustained-release ocular delivery system for pefloxacin mesylate using rabbit eye model

The major purpose of this study was to develop and characterize a series of carbopol- and methyl cellulose-based solutions as the in situ gelling vehicles for ophthalmic drug delivery. The rheological properties, in vitro release as well as in vivo pharmacological response of a combination of polymer solutions, including carbopol and methyl cellulose, were evaluated. It was found that the optimum concentration of carbopol solution for the in situ gel-forming delivery systems was 0.3% (w/w), and that for methyl cellulose solution was 1.5% (w/w). The mixture of 0.3% carbopol and 1.5% methyl cellulose solutions showed a significant enhancement in gel strength in the physiological condition; this gel mixture was also found to be free flowing at pH 4.0 and 25 degrees C. The rheological behaviors of carbopol/methyl cellulose solution were not affected by the incorporation of the drug. Drug levels in the aqueous humor of the rabbits were well above the MIC-values of relevant bacteria after 12 hours, the results of an optimized formulation containing 0.18% of pefloxacin mesylate compared well with the 0.3% marketed eye drop formulation, indicating our formulation to be significantly better considering that a similar effect was obtained at half the concentration. Both the in vitro release and in vivo pharmacological studies indicated that the carbopol/methyl cellulose solution had better ability to retain drug than did the carbopol or methyl cellulose solutions alone. The results demonstrated that the carbopol/methyl cellulose mixture can be used as an in situ gelling vehicle to enhance the ocular bioavailability of pefloxacin mesylate.     

Transfersomes: A Novel Vesicular Carrier for Enhanced Transdermal Delivery of Sertraline: Development,Characterization, and Performance Evaluation

The aim of the present study was to investigate transfersomes as a transdermal delivery system for the poorly soluble drug, sertraline, in order to overcome the troubles associated with its oral delivery. Different transfersomal formulations were prepared with non-ionic surfactant (span 80), soya lecithin, and carbopol 940 by the rotary evaporation sonication method. The prepared formulations were characterized for light microscopy, particle size analysis, drug entrapment, turbidity, drug content, rheological studies, in vitro release, ex vivo permeation, and stability studies. The optimized formulation was evaluated for in vivo studies using the modified forced swim model test. FTIR studies showed compatibility of the drug with excipients. The result revealed that sertraline in all of the formulations was successfully entrapped with uniform drug content. Transfersomal gel containing 1.6% of the drug and 20% of span 80 was concluded to be the optimized formulation (EL-SP4), as it showed maximum drug entrapment (90.4±0.15%) and cumulative percent drug release(73.8%). The ex vivo permeation profile of EL-SP4 was compared with the transfersomal suspension, control gel, and drug solution. The transfersomal gel showed a significantly higher (p<0.05) cumulative amount of drug permeation and flux along with lower lag time than the drug solution and drug gel. It also owed to better applicability due to the higher viscosity imparted by the gel rather than the transfersomal suspension, and no skin irritation was observed. The modified forced swim test in mice revealed that the transfersomal gel had better antidepressant activity as compared to the control gel. Thus, the study substantiated that the transfersomal gel can be used as a feasible alternative to the conventional formulations of sertraline with advanced permeation characteristics for transdermal application.     

Quantitation of low level unconjugated polysaccharide in tetanus toxoid-conjugate vaccine by HPAEC/PAD following rapid separation by deoxycholate/HCl

A simple and rapid acid precipitation method has been applied successfully for separating free capsular polysaccharide of Haemophilus influenzae type b (polyribosyl ribitol phosphate, PRP) from PRP–tetanus toxoid conjugate (PRP-T) in a final dosage amount of low-level materials. The unconjugated PRP was found to stay in the supernatant without precipitation, while conjugated PRP-T was fully precipitated. High performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) has been applied for analysis of the PRP content in the supernatant after the separation. This method requires minimum sample handling and is specific, sensitive and reproducible making it suitable for release and stability testing of PRP-T in final containers.     

喷昔洛韦眼用温度敏感原位凝胶的制备及评价（英文）

目的研制以普朗尼克F127为主要基质的喷昔洛韦制剂,以提高其眼部生物利用度。方法通过将HPMC K4M或卡波姆934P与普朗尼克F127复合使用,制备了喷昔洛韦的温度敏感原位凝胶。以胶凝温度、流变学、药物释放特性、药代动力学及眼部刺激性等为指标进行筛选,得到最优化处方。结果使用HPMC K4M或者卡波姆934P均能降低凝胶的胶凝温度,略微增加其粘度,延缓体系中药物的释放速率;药物释放为非Fick扩散;所有处方均未表现出眼部刺激或对角膜的损伤;含卡波姆934P和普朗尼克F127的凝胶体系的眼部生物利用度最高。结论含普朗尼克F127的喷昔洛韦制剂能够以滴眼液的形式给药,而达到眼部温度时可形成凝胶;体内外评价结果表明,含有HPMC K4M或卡波姆934P以及低浓度普朗尼克F127（12%）的喷昔洛韦制剂,提高了药物在眼部的生物利用度,是一种很有前景的眼部给药系统。     

人凝血因子Ⅷ提取工艺优化研究

目的：优化血浆中人凝血因子Ⅷ提取工艺条件。方法以人血浆冷沉淀为原料,人凝血因子Ⅷ的比活性回收率作为评价指标,采用单因素试验,对铝胶的加量,铝胶吸附时的pH值、酸沉淀时的pH值等进行研究,优选提取工艺。结果加入冷沉淀重量12%的铝胶,并将pH值调节至7.1进行吸附,上清液中人凝血因子Ⅷ比活性可达9.50IU/mg,酸沉淀时将pH值调节至6.3,离心上清夜中人凝血因子Ⅷ比活性为11.50 IU/mg。经过以上两步处理,可将人凝血因子Ⅷ比活性提高近2倍。结论人凝血因子Ⅷ提取工艺优化研究工艺合理、可行,能有效达到纯化提取人凝血因子Ⅷ的效果。