新型水包油佐剂SP01的安全性评价

目的通过动物实验评价新型水包油佐剂SP01的安全性。方法通过昆明小鼠安全药理试验、昆明小鼠急性毒性试验、Hartley豚鼠过敏反应试验、溶血性试验、日本大白兔局部刺激性试验、SD大鼠长期毒性试验及BALB/c小鼠致突变、致畸、致瘤试验,对水包油佐剂SP01的安全性进行全面评价。结果水包油佐剂SP01对昆明小鼠的自主活动无明显影响;急性毒性试验中可观察到水包油佐剂SP01组昆明小鼠给药后无死亡,体重明显增加,且各组织器官无病理变化;水包油佐剂SP01注射豚鼠未见任何过敏反应及症状;溶血性试验结果显示,水包油佐剂SP01在体外对兔红细胞无溶血作用,不引起红细胞凝聚;将水包油佐剂SP01经日本大白兔耳缘静脉注射连续刺激5 d后,佐剂组与氯化钠注射液组家兔耳缘静脉给药部位肉眼观察均无明显变化,病理切片显微镜观察均未见炎性浸润、血管周围组织出血及坏死等刺激现象;将水包油佐剂SP01经肌肉连续注射大鼠1个月,佐剂组大鼠体重明显增加,精神状态、食欲、睡眠情况与氯化钠注射液组无明显差异,停药后连续观察3个月,大鼠精神状态、食欲、睡眠情况均良好;将水包油佐剂SP01连续经BALB/c小鼠腿部肌肉注射14 d后,其肌肉组织病理切片显示,与氯化钠注射液组小鼠的肌肉组织无明显差异。结论水包油佐剂SP01对实验动物无急性毒性、过敏反应、局部性刺激、长期毒性及致突变、致畸、致瘤作用,是安全、可应用的油类佐剂,本研究为该系列水包油佐剂的临床应用提供了实验依据。     

LT70-DPC结核亚单位疫苗安全性的初步评价

目的初步评价以阳离子脂质体二甲基三十六烷基铵(dimo-thylidioctyl ammonium bro-mide,DDA)、聚肌胞苷酸(Poly I∶C)及胆固醇复合佐剂(简称为DPC)为佐剂的结核融合蛋白ESAT6-Ag85B-MPT64(190-198)-Mtb8.4-Rv2626c(LT70)亚单位疫苗的安全性。方法急性毒性试验:分别将低剂量与高剂量(分别相当于人用剂量的10 000和40 000倍)的LT70-DPC结核亚单位疫苗经皮下、腹腔两种途径免疫小鼠,观察小鼠毒性反应、死亡情况及体重变化等,同时推算小鼠对LT70-DPC亚单位疫苗的最大耐受剂量(maximum tolerated dose,MTD);异常毒性试验:将小鼠和豚鼠分别经腹腔注射LT70-DPC结核亚单位疫苗,观察动物异常反应、死亡情况及体重变化等;全身过敏试验:豚鼠经LT70-DPC结核亚单位疫苗免疫后,经相应疫苗静脉攻击,观察豚鼠过敏反应。结果急性毒性试验:皮下及腹腔注射后14 d内均未见小鼠的异常反应和死亡,体重均增加,小鼠对该疫苗的MTD为:LT70蛋白6 600μg/kg,DDA166 600μg/kg,Poly I∶C33 200μg/kg,胆固醇50 600μg/kg;异常毒性试验:注射后7 d内,小鼠及豚鼠全部健存,均未出现异常症状,体重增加;全身过敏试验:攻击后1 h内,豚鼠均未出现过敏反应。结论 LT70-DPC结核亚单位疫苗具有良好的安全性。     

鱼精蛋白对PLA微球佐剂的调控

采用快速膜乳化法,以聚乳酸(PLA)为材料,分别以乙酸乙酯和二氯甲烷为有机溶剂,制备了粒径400和900 nm的均一PLA微球和鱼精蛋白(PS)修饰的PLA-PS微球,以微球为佐剂、HBsAg为抗原免疫小鼠,在动物水平上研究其免疫调控作用,考察了不同粒径微球对抗原体内免疫效果的影响. 结果表明,鱼精蛋白修饰后的PLA微球对乙肝表面抗原的吸附率增加,未修饰的PLA微球对抗原的吸附率仅为2.07%,修饰后吸附率为8.16%;修饰后微球为佐剂的免疫效果优于未修饰微球为佐剂的免疫效果,鱼精蛋白修饰能显著提升小鼠脾细胞分泌Th1型细胞因子(IL-2和IFN-γ)数(分别提升了261%和139%),分泌Th2型细胞因子IL-4数增加了67%;用修饰后平均粒径400 nm微球为佐剂免疫小鼠,IL-2分泌水平是900 nm微球为佐剂的1.68倍.     

艾纳香口腔护理液安全性评价研究

选用SD大鼠进行口腔黏膜刺激性试验、口腔及黏膜用药急性毒性试验;选用白色豚鼠进行过敏试验;选用昆明种小鼠进行灌胃给药急性毒性试验,观察给药后动物口腔黏膜、受药皮肤、体重变化、全身毒性反应情况及死亡只数。艾纳香口腔护理液在口腔黏膜刺激性试验中,未发现SD大鼠口腔黏膜组织异常;在过敏试验中,白色豚鼠皮肤用药未引起过敏反应;在急性毒性试验中,大鼠的耐受性良好;小鼠灌胃时测得的最大给药量(MLD)为288 mg/kg,相对于成人给药量的1 200倍。上述动物均未见死亡和明显毒性反应。以上试验说明艾纳香口腔护理液无口腔黏膜刺激性和致敏性,无明显急性毒性反应,提示其作为口腔外用护理液临床用药安全。     

壳聚糖微球及其与白油混合乳液对禽用新流二联疫苗佐剂的免疫效果

用不同粒径的壳聚糖季铵盐凝胶微球及其与白油的混合乳液吸附新城疫及禽流感灭活抗原,评价其对新流二联灭活疫苗的免疫增强效果.结果表明,吸附时间2 h、微球浓度2 mg/m L、微球粒径1μm为吸附新流二联灭活抗原的最佳条件,抗原吸附率达90%.免疫SPF鸡后,壳聚糖季铵盐微球及混合乳液均能显著提高新城疫及禽流感病毒特异性抗体效价,且混合乳液组显著优于微球组.与白油佐剂相比,壳聚糖季铵盐微球及混合乳液均能显著增强细胞免疫,淋巴细胞增殖水平为纯抗原组的1.5倍,具有用作禽类疫苗佐剂的巨大潜力.     

杀螨脒的毒性研究

98%纯度的原药大、小鼠急性经口LD_(50)为104(雌)、129(雄)和145(雄)、151(雌)毫克/公斤;雄大鼠经皮MLD>2克/公斤,药剂对家兔皮肤和眼无刺激/腐蚀作用;对豚鼠皮肤无致敏作用,大鼠蓄积毒性属弱蓄积性;大鼠致畸试验未发现胚胎毒性和致畸作用。由Ames试验、小鼠骨髓细胞微核试验和染色体畸变试验及睾丸初级精母细胞染色体畸变试验所组成的致突变测试系统,检测结果未发现诱变作用。     

人用重组禽流感血凝素疫苗接种大鼠的安全性及免疫原性

目的评价家蚕生物反应器生产的重组禽流感血凝素疫苗的安全性和免疫原性。方法将SD大鼠随机分为5组,分别经皮下注射9.4、1.88和0.375mg/kg剂量的重组禽流感血凝素疫苗(含氢氧化铝佐剂)、A(lOH)3和生理盐水,每10d注射1次,连续注射3次,分别于第1次给药后第2、22和42天进行血液学、血液生化学、病理学和免疫原性检测。结果3个剂量组疫苗第1次给药后第42天,检测抗体滴度(P/N)在8～11之间,CD4+、CD8+T淋巴细胞含量及IFNγ和IL-2的含量与两对照组相比,未见明显升高;连续3次给药后,大鼠的血液学和生化学指标均无明显改变;给药期间注射部位皮下出现肉芽肿样结节,经20d恢复后有所改善;9.4和1.88mg/kg剂量组动物出现可逆性的脾肿大症状;整个实验期间,动物未出现任何其他明显不良反应。结论重组禽流感血凝素疫苗对大鼠具有良好的安全性,但免疫原性较低。     

15价肺炎球菌结合疫苗动物急性、长期毒性及免疫原性

目的 评价15价肺炎球菌结合疫苗（15-valent pneumococcal conjugate vaccine,PCV15）的动物急性、长期毒性及免疫原性。方法 将60只ICR小鼠随机分为磷酸铝佐剂对照组、0.9%NaCl对照组和2人份剂量PCV15组,每组20只,雌雄各半,进行急性毒性试验;将90只SD大鼠随机分为磷酸铝佐剂对照组、0.9%NaCl对照组和1人份剂量PCV15组,每组30只,雌雄各半,共免疫5次,间隔28 d,进行长期毒性试验;将45只NIH小鼠随机分为0.9%NaCl组、1/4人用剂量的市售13价肺炎球菌结合疫苗（PCV13）组和1/4人用剂量的PCV15组,每组15只,雌性,共免疫3次,间隔2周,进行免疫原性试验及免疫过程中安全性观察,全程监测并记录小鼠的体质量、体征及行为变化,并在每次免疫2周后经眼眶采血,间接ELISA检测血清中抗各型肺炎链球菌荚膜多糖抗体水平。结果 急性毒性试验表明,注射2人份剂量PCV15后,小鼠均未出现明显异常反应或死亡。长期毒性试验显示,1人份剂量的PCV15对大鼠临床症状、血液学和血清生化指标无明显影响,各器官未见明显病理学异常。...     

竹叶黄酮壳聚糖缓释微球的制备及其性能研究

以壳聚糖为载体材料,戊二醛为交联剂,竹叶黄酮作为模型药物,采用乳化交联法,制备了载竹叶黄酮壳聚糖缓释微球。研究了载药比对微球的形貌、药物收率、包埋率和载药量的影响。结果表明:竹叶黄酮-壳聚糖微球呈规则球形,粒径10~50μm;竹叶黄酮经壳聚糖加载后,其释放时间长,释药均匀,具有良好的缓释性能。     

代森环毒性实验研究

代森环对Wistar大鼠的急性经口LD_(50)为4640毫克/公斤,急性经皮LD_(50)<10000毫克/公斤,有强蓄积毒性。对家兔皮肤、眼粘膜无明显刺激作用。90天亚慢性毒性实验63毫克/公斤为无作用剂量,病理形态学检查未见明显的病理改变。Ames实验,大鼠睾丸精母细胞染色体畸变实验、小鼠骨髓细胞微核实验均为阴性结果,故无致突变作用。     

人用皮内注射布氏菌活疫苗毒理及药效学研究

目的研究人用皮内注射布氏菌活疫苗的毒理及药效学作用,以评价其可行性。方法对布氏菌活疫苗进行小白鼠急性毒性试验、局部刺激试验、全身过敏试验、长期毒性试验及豚鼠皮肤变态反应试验并免疫小鼠,进行血清抗体检测。结果小鼠皮内注射布氏菌活疫苗的最大耐受量为1·0×109个菌,用布氏菌素注射足底,可引起明显的迟发型变态反应。局部刺激试验,只有注射1·0×109个菌的部位会在48h后化脓,但在1~2周内可自愈。全身过敏试验均未出现任何异常反应,毒性试验、各项血常规、血生化检查均未见显著差异,各脏器组织的病理学检查亦未见病理改变。免疫12周后,对豚鼠骨髓造血细胞亦无影响。对皮内免疫12周的豚鼠作背部皮肤变态反应试验,24~48h结果为阳性。皮内免疫小鼠14d后可测得小鼠体内有抗体产生,50d后测得小鼠体内抗体均呈强阳性,对照组小鼠14d与50d抗体检测均为阴性。结论毒理和药效学试验结果显示,布氏菌活疫苗用皮内注射的途径进行免疫是可行的。     

表达狂犬病毒糖蛋白的重组痘苗病毒对啮齿动物接种的反应

狂犬病毒糖蛋白重组痘菌病毒（V－RG）与非重组痘菌病毒比较，V－RG对小白鼠爆发性致死量下降10000LD50；对家兔眼粘膜及皮肤感染后，3～5天内呈现轻微的红、肿，3周内全部消退。所有局部损害仅限于表皮，未见其它不良反应。V－RG经口免疫的动物7天即产生抗体，于第14天能抵抗致死量狂犬病毒脑内或肌肉内攻击。     

缓释破伤风类毒素微球的制备及其免疫原性

目的 用可生物降解缓释微球,单剂投递破伤风类毒毒。方法 用溶剂挥发法,将破伤风类毒素(TT)包入两种丙交酯-乙交酯共聚物制备成微球,用电镜观察微球的粒经与表面形态,用Bradford法及SDS-PAGE检测微球内蛋白含量及抗原结构的变化。TT微球免疫豚鼠后,观察抗体应答。结果 微球表面光滑,成球规律,两种微球平均粒径为8.4μm及9.7μm,包裹率为48％及56％,包裹后的TT结构未改变。TT微球免疫豚鼠后,1针诱导的抗TT-IgG及中和抗体滴度与3针铝吸附TT相似;TT微球免疫血清及铝吸附TT免疫血清与游离破伤风毒素有相似的结合特性。加强免疫后,TT微球的回忆应答优于铝佐剂疫苗。结论 可生物降解缓释微球单剂投递破伤风类毒素,可产生持续高的免疫应答。     

A Lower Irradiation Dose of 308 nm Monochromatic Excimer Light Might Be Sufficient for Vitiligo Treatment: A Novel Insight Gained from In Vitro and In Vivo Analyses

A 308 nm monochromatic excimer light (MEL) is widely used to treat patients with vitiligo. However, dose optimization still needs to be clarified. This study aimed to obtain objective evidence regarding various doses of MEL irradiation, induced cell level changes in vitro, and skin level alterations in vivo. Cultured human keratinocytes were irradiated with MEL using various doses. After irradiation at low doses, stem cell factor, endothelin-1, and glycoprotein nonmetastatic melanoma protein B, factors that activate and protect melanocytes, were found to be significantly elevated in keratinocytes. After irradiation using medium and high doses, inflammatory cytokines were induced. The amount of ATP released and the level of inflammasome activation, which are known to be related to interleukin-1β activation, were also increased. The back skin of guinea pigs and mice were irradiated with MEL at varying doses. After irradiation, an increase of epidermal melanin and epidermal melanocytes was confirmed, using the minimal erythemal dose or less. In rhododendrol-induced leukoderma guinea pigs, a much lower dose of MEL irradiation was effective, when compared with the effective dose for control guinea pigs. Our results suggest that a lower irradiation dose of MEL might be sufficient and more suitable for repigmentation in vitiligo treatment.     

流感亚单位疫苗裂解剂残留量检测及安全性试验

目的检测流感亚单位疫苗的裂解剂残留量,并观察其安全性。方法制备3批流感亚单位疫苗,采用比色法检测其裂解剂Tween-80和CTAB的残留量,通过动物实验观察其安全性。结果制备的3批疫苗样品,Tween-80的残留量平均在68.3~72.0μg/ml之间,CTAB残留量平均在36.3~38.7μg/ml之间。在安全性试验中,豚鼠未出现过敏反应,小鼠和豚鼠未出现异常毒性反应。结论制备的流感亚单位疫苗的裂解剂残留量符合拟定规程要求,安全性良好。     

Phytosterols and pectin added to a high‐saturated fat diet do not show hypocholesterolemic activity in female guinea pigs

This paper presents the results of a study whose aim was to test the effects of several doses of pectin and phytosterols on the sterol content in plasma, liver and feces of guinea pigs, when added to a high‐saturated fat diet. The treatments followed a 3×3 factorial design, with three levels of pectin (0, 3.67 and 6.93%) and three levels of phytosterols (0, 1.37 and 2.45%). Seventy‐two female Dunkin Hartley guinea pigs were randomly assigned to the treatment groups (eight animals per group). The duration of the treatment was 4 weeks. No differences were found in plasma cholesterol concentrations, while in liver we saw a reduction in cholesterol concentration after phytosterol feeding. Moreover, we found no pectin effects. Plant sterol concentration increased in plasma and liver after phytosterol ingestion, with the highest concentrations being obtained with the intermediate pectin dose. Our results suggest that a high‐saturated diet may impair the cholesterol‐lowering properties of plant sterols and pectin.     

The Safety Evaluation of Salvianolic Acid B and Ginsenoside Rg1 Combination on Mice

Our previous study indicated that the combination of salvianolic acid B (SalB) and ginsenoside Rg1 (Rg1), the main components of Salvia miltiorrhizae and Panax notoginseng, improves myocardium structure and ventricular function in rats with ischemia/reperfusion injury. The present study aimed to determine the safety of the combined SalB and Rg1 (SalB-Rg1) in mice. The safety of SalB-Rg1 was evaluated through acute toxicity and repeated-dose toxicity. In the acute toxicity study, the up and down procedure was carried out firstly, and then, the Bliss method was applied. In the toxicity study for seven-day repeated treatment of SalB-Rg1, forty Kunming mice were randomly divided into four groups. The intravenous median lethal dose (LD₅₀) of the SalB-Rg1 combination was 1747 mg/kg using the Bliss method. For both the acute toxicity study and the seven-day repeated toxicity study, SalB-Rg1 did not induce significant abnormality on brain, heart, kidney, liver and lung structure at any dose based on H&E stain. There were no significant changes related to the SalB-Rg1 toxicity detected on biochemical parameters for two kinds of toxicity studies. The LD₅₀ in mice was 1747 mg/kg, which was more than one hundred times higher than the effective dose. Both studies of acute toxicity and seven-day repeated dose toxicity indicated the safety of the SalB-Rg1 combination.     

Intravenous Single-Dose Toxicity of Redaporfin-Based Photodynamic Therapy in Rodents

We assessed the tolerability and safety in rodents of a single intravenous (i.v.) dose of redaporfin, a novel photosensitizer for Photodynamic Therapy (PDT) of cancer. Two approaches were used to evaluate acute toxicity: (i) a dose escalation study in BALB/c mice to evaluate the maximum tolerated dose of redaporfin; and (ii) a safety toxicology study in Wistar rats, of a single dose of redaporfin, with or without illumination, to evaluate possible signs of systemic toxicity. Redaporfin formulation was well tolerated by mice, with no signs of adverse reactions up to 75 mg/kg. In rats, there were no relevant changes, except for a significant, but transient, increase in the blood serum markers for hepatic function and muscle integrity, and also on neutrophil counts, observed after the application of light. The overall results showed that redaporfin-PDT is very well tolerated. No abnormalities were observed, including reactions at the injection site or skin phototoxicity, although the animals were maintained in normal indoor lighting. Redaporfin also showed a high efficacy in the treatment of male BALB/c mice with subcutaneously implanted colon (CT26) tumours. Vascular-PDT with 1.5 mg/kg redaporfin and a light dose of 74 J/cm² led to the complete tumour regression in 83% of the mice.     

快速膜乳化与热固化法制备粒径均一的pH敏感壳聚糖季铵盐凝胶微球

以具有升温自固化特性的壳聚糖季铵盐/甘油磷酸钠混合溶液为水相,利用快速膜乳化与热固化法制备了粒径均一、pH敏感的壳聚糖季铵盐凝胶微球,考察了跨膜压力、水油相组成、水油相体积比及微孔膜孔径等对微球粒径、结构和药物包埋率的影响. 结果表明,得到粒径698±57.33, 1145±71.48, 2021±53.63及3984±191.72 nm、粒径分布窄(多分散系数<0.1)、药物包埋率高达75.49%±2.62%的凝胶微球. 所制微球生物相容性好,有明显的pH敏感性,中性和碱性环境下结构稳定,药物缓释,pH=7.4时24 h内药物累计释放率为34.6%;酸性环境下微球崩解,药物快速释放,pH=5.5时1 h内药物累计释放率高达79.6%.     

壳聚糖季铵盐表面修饰对载紫杉醇PLGA微球体外药效学的影响

采用聚(乳酸-羟基乙酸)共聚物(PLGA)纳微球装载紫杉醇,并用壳聚糖季铵盐(HTCC)对PLGA微球表面进行镀层修饰,比较了修饰前后载药微球的形貌、粒径、电位、载药率、释药行为和细胞杀伤效果. 结果表明,修饰后微球表面圆整光滑,平均粒径为882 nm,载药率可达5.15%,包埋率达70.46%,体外释药22 d累积释药率为70.17%,与修饰前没有显著性差异;但修饰后微球表面电荷由修饰前的-14.8 mV翻转为+36.7 mV,肿瘤细胞对PLGA和HTCC-PLGA载药微球的内吞量分别是Taxol?的5.6和9.7倍,且HTCC-PLGA载药微球对细胞杀伤效果显著,是一种有潜力的难溶性药物递送系统.