咪唑苯脲注射液特殊安全性试验研究

通过豚鼠、新西兰大白兔的皮肤致敏、皮肤刺激、肌肉刺激、红细胞溶血试验,考察咪唑苯脲注射液的安全性。试验用新西兰大白兔6只进行皮肤刺激,染毒剂量为咪唑苯脲注射液0.5 mL/只(规格:1211.5 mg/mL);用白化豚鼠进行皮肤致敏试验,按0.5 mL/kg剂量,以豚鼠背部左侧皮肤进行染毒;用新西兰大白兔8只,进行肌肉刺激试验,在右侧股四头肌注入0.1、0.2、0.5 mL/kg bw,左侧股四头肌注入同样体积的灭菌0.9%氯化钠溶液作对照;用新西兰大白兔2只,进行红细胞溶血试验,采用体外试管法进行,采心脏血50 mL,制成红细胞悬液,加入受试药物原液温育3 h。结果显示,皮肤刺激试验:受试物咪唑苯脲注射液各个时间点的刺激反应积分均值为0;皮肤致敏试验:受试药物组和阴性对照组豚鼠的皮肤过敏反应率为0;肌肉刺激试验:给予受试物各个时间点的刺激反应积分均值为0;红细胞溶血试验:受试药物及阴性对照在3 h内红细胞全部下沉,上清液体均为澄明,溶液中未见棕红色或红棕色絮状沉淀。试验表明,咪唑苯脲注射剂无皮肤刺激性、不出现过敏反应、无肌肉刺激性、无溶血和凝聚作用,临床上可以肌肉注射使用。     

催奶灵注射液的安全性试验

通过豚鼠的全身主动过敏性试验、家兔的血管和肌肉局部刺激性以及体外溶血性试验,对催奶灵注射液进行了安全性评价。结果表明：本品对豚鼠无明显过敏性反应,静脉和肌肉注射对家兔耳缘静脉和肌肉组织无明显局部刺激,对家兔红细胞无明显体外溶血,各项安全性均较好,可推荐临床使用。     

加米霉素注射液特殊安全性试验研究

通过豚鼠、大白兔的皮肤致敏、皮肤刺激、肌肉刺激、红细胞溶血试验,考察加米霉素注射液的安全性。试验用新西兰大白兔8只进行皮肤刺激,染毒剂量为加米霉素注射液0.5 mL/只(规格:150 mg/mL);用白化豚鼠进行皮肤致敏试验,按0.5 mL/kg剂量,以豚鼠背部左侧皮肤进行染毒;用新西兰大白兔24只,进行肌肉刺激试验,在右侧股四头肌注入6、12、30 mg/kg bw,左侧股四头肌注入同样体积的灭菌0.9%氯化钠溶液作对照;用新西兰大白兔2只,进行红细胞溶血试验,采用体外试管法进行,采心脏血50 mL,制成血细胞悬液,加入受试药物原液温育3 h。结果显示,皮肤刺激试验:受试物加米霉素注射液各个时间点的刺激反应积分均值为0;皮肤致敏试验:受试药物组和阴性对照组豚鼠的皮肤过敏反应率为0;肌肉刺激试验:给予受试物各个时间点的刺激反应积分均值为0;红细胞溶血试验:受试药物及阴性对照在3 h内红细胞全部下沉,上清液体均为澄明,溶液中未见棕红色或红棕色絮状沉淀。试验表明加米霉素注射剂无皮肤刺激性、不出现过敏反应、无肌肉刺激性、无溶血和凝聚作用,临床上可以肌肉注射使用。     

Feeding of Hartley Guinea Pigs for Experiment and Establishment of Allergic Rhinitis Model of Nasal Drops

The assay was aimed to explore the feeding and the best method to establish allergic rhinitis (AR) model in Hartley guinea pigs.Using the single factor experiment, the assay studied the effects of animal species, feeding way, ovalbumin (OVA) sensitization dose, time for AR model, and observed allergic reaction symptoms and nasal mucosal pathological section of Hartley guinea pig.The results showed that Hartley guinea pig was the best experimental animal for the AR experiment of nasal drops;Feed was ordinary mice feed + lettuce leaves or leaf lettuce;OVA as the sensitizer, concentration of basic sensitization was 0.1 to 0.3 mg/mL OVA, the next day intraperitoneal injection 1 time, for 7 times;Concentration of stimulated sensitization was 2% OVA, 10 to 30 μL each nostril, 1 time a day, after 6 to 9 d, compared with the blank group, the Hartley guinea pig model group had significant allergic symptoms and nasal mucosa lesions.This modeling method was simple and economic, good reproducibility, and could be used as the best method for AR model establishment of nasal drops.     

试验用Hartley豚鼠的饲养及滴鼻剂变应性鼻炎建模

为探索试验用Hartley豚鼠的饲养及建立滴鼻剂变应性鼻炎(AR)模型的有效方法,本试验采用单因素试验,考察了试验动物种类、饲养方式、卵清蛋白(OVA)致敏剂量、时间对滴鼻剂AR建模的影响,观察空白组与模型组Hartley豚鼠过敏反应症状和鼻黏膜病理切片。结果显示,就滴鼻剂的AR试验而言,Hartley豚鼠是较好的试验动物;饲料为普通小鼠饲料+莴苣叶或油麦菜;OVA为致敏剂,基础致敏浓度为0.1~0.3 mg/mL OVA,隔天腹腔注射1次,共7次;激发致敏浓度为2%,两侧鼻腔各10~30 μL,每天1次,6~9 d后与空白组相比,模型组Hartley豚鼠有明显的过敏反应症状和鼻黏膜病变。该建模方法简单经济,重现性好,可作为滴鼻剂AR建模的最佳方法。     

Study on Sterility Test Method of Ivermectin Microemulsion

The assay was aimed to study the sterility test and validation test of ivermectin microemulsion preparation and establish a sterility test method for ivermectin microemulsion preparation.The test method was carried out according to the method in volumeⅠ, Chinese Veterinary Pharmacopoeia Edition 2005.By choosing positive control bacteria and defining washing volumes in sterility test, the membrane-filter method was used to test the quantity of 10 bottles of test samples, and the sterility test was established.The result of method validation test showed that the test and all of positive control bacteria and microorganism growth after each filter being washed with 400 mL 0.1% peptone solution.It illustrated that the samples had no antimicrobial activity under the sample quantity and test condition.This method was available for sterility test of ivermectin microemulsion preparation.Using this method to test three lots test samples, the results showed that the positive control bacteria grew well within 24 h.The negative control bacteria and three lots test samples were sterile.It indicated that sterility test results met the requirements.     

伊维菌素微乳无菌检查方法学的研究

本试验旨在对伊维菌素微乳制剂进行无菌检查方法学验证和无菌检查试验,确认本试验所用的方法适用于该制剂的无菌检查。按《中国兽药典》2005版一部(附录118)所载"无菌检查法"项下进行试验,通过对阳性对照菌、不同量冲洗液等条件的选择,采用薄膜过滤法对10瓶供试品(每种试验菌的样品量)进行检测,建立了无菌检查方法。经方法验证,用400mL 0.1%蛋白胨水溶液冲洗后,含供试品容器中的7个阳性菌试验组与阳性菌对照组相比均生长良好,说明供试品的该检验量在该检验条件下无抑菌作用或其抑菌作用可以忽略不计,可以用该方法进行供试品的无菌检查。对3批供试品进行无菌检查,阳性对照菌均在24h内生长良好,阴性对照均澄清,无菌生长,3批供试品均澄清,无菌生长,无菌检查试验结果符合规定。     

Content Determination of Ivermectin in the Ivermectin Microemulsion Injection

In order to establish the determination method of ivermectin (IVM) in the ivermectin microemulsion injection by high performance liquid chromatography (HPLC), Hypersil ODS2 colunm (5 μm,4.6 mm×250 mm) was used in this study. The mobile phase composed of methanol, acetonitrile and water (35:60:5,V/V/V) at a flow rate of 1 mL/min. The detection wave length was set at 244 nm and the column temperature was 30 ℃. The results showed that the HPLC system suitability of IVM was good. A good linear correlation of IVM was observed within the concentration range 80 to 320 μg/mL, and the average recovery rate was 101.90%±2.94% with RSD was 2.88%, the regression equation was Y=22 700X+2 510 (R²=0.9998). The RSD of IVM content in ivermectin microemulsion injection was 1.86%. The method was accurate and reliable,reproducible,easy to operate, which could be used in new type of ivermectin microemulsion injection. The method could be used as the basis of quality control and establishing a quality standard, and to provide basis for quality evaluation, also can provide reliable reference for safe veterinary clinical application in the future.     

伊维菌素微乳中伊维菌素的含量测定方法研究

为建立伊维菌素微乳中伊维菌素含量的高效液相色谱（HPLC）测定方法,选用Hypersil ODS2 （5 μm,4.6 mm×250 mm）色谱柱,流动相为甲醇∶乙腈∶水为35∶60∶5（V/V/V）,检测波长为244 nm,柱温为30 ℃,流速为1 mL/min进行测定。结果显示,伊维菌素在该色谱条件下,系统适应性良好,在80~320 μg/mL浓度范围内线性关系良好,回归方程为：Y=22 700X+2 510,R²=0.9998,总平均回收率为101.90%±2.94%,RSD为2.88%,对中试生产的3批伊维菌素微乳进行含量测定,RSD为1.86%。表明该含量测定方法准确可靠,重现性好,可用于伊维菌素微乳中伊维菌素含量的测定,并为该新型制剂的质量标准的制定和质量评价提供依据,也为后期的临床安全应用提供可靠的参考。     

氟苯尼考悬乳剂的安全试验研究

通过试验研究对氟苯尼考悬乳剂的安全性进行评价。采用豚鼠和家兔分别进行本品的全身用药过敏性和肌肉局部刺激性、红细胞体外溶血性试验。结果显示本品临床给药剂量对豚鼠无全身过敏性、对家兔肌肉注射给药无明显局部刺激性、对家兔红细胞无体外溶血性。表明本品的安全性良好,为实际应用提供了依据。     

复方环丙沙星乳剂安全性试验研究

采用动物药物安全性评价试验的方法,对豚鼠、家兔等动物进行了复方环丙沙星乳剂的热源试验、体外红细胞溶血性试验、局部刺激性试验及全身主动过敏试验。结果显示,复方环丙沙星乳剂无热原性、体外红细胞溶血性和过敏性,也几乎无明显局部刺激性。表明该制剂的安全性良好,为其在兽医临床上的实际应用提供了依据。     

Supplementation of heterologous complement induces anti-Thy-1.1 nephritis in the Mongolian gerbil (Meriones unguiculatus)

Anti-Thy-1.1 nephritis in the rat is a popular experimental model for mesangial proliferative glomerulonephritis (GN). This model is characterized by direct binding of anti-Thy-1.1 antibody with Thy-1.1 antigen expressed on mesangial cells (MCs) of glomeruli in the rat. A single injection of anti-rat thymocyte serum (ARTS) results in GN with proteinuria and extensive mesangiolysis. Development of mesangiolysis and proteinuria are complement-dependent. We previously demonstrated Thy-1.1 antigen, similar to the rat, in thymocytes, brain cells and MCs of the kidney in the Mongolian gerbil (MG). In this study, we attempted to develop a MG nephritis model, but an injection of ARTS did not induce GN. An additional injection of guinea pig serum as a complement after ARTS injection resulted in anti-Thy-1.1 nephritis in MG. Degeneration of MCs and neutrophil infiltration were observed 1 hr after GP serum injection. Mesangiolysis and fibrin exudation occurred 12 hr after the injection and MC proliferation was apparent 7 days after the injection. In the complement-dependent hemolytic test, MG serum could not hemolyze sheep erythrocytes. These results suggested low activity, or depletion of some factors, in complements of MG serum.     

防治犬真菌性皮肤病中药软膏剂的制备及质量评价

为寻找安全有效防治犬真菌性皮肤病的临床药物,采用乳化法制备出防治犬真菌性皮肤病的复方中药软膏剂,并通过对中药软膏的外观性状、稳定性、刺激性、局部致敏性等质量标准进行综合评价。试验结果表明,本试验制备的中药软膏剂外观细腻,质地均匀,质量稳定,无明显急性毒性反应、无刺激性、不产生局部及全身过敏反应。     

Lactate dehydrogenase and creatine phosphokinase isoenzymes in tissues of laboratory animals

The lactate dehydrogenase (LDH) and creatine phosphokinase (CPK) isoenzyme distributions in tissues of the ICR mouse, Wistar rat, guinea pig and golden hamster were analyzed by histoelectrophoresis. Tissues obtained were as follows: liver, pancreas, stomach, small intestine, heart, femoral muscle, uterus, kidney, spleen, lymph node, cerebrum, spinal cord and erythrocyte. Histoelectrophoresis was for the direct analysis of LDH and CPK isoenzymes in the tissues and had high practical value compared with previous tissue-extraction methods. In tissues of the mouse, guinea pig and golden hamster, LDH isoenzymes showed five bands. In the rat, LDH isoenzyme was separated into four fractions. CPK isoenzyme showed three bands; BB, MB and MM. In some tissues, the MM band was separated into two sub fractions.     

多拉菌素浇泼剂临床前毒理学试验研究

观察多拉菌素浇泼剂对动物的安全药理学和皮肤的毒性作用。用家兔进行皮肤安全药理学试验,用豚鼠进行皮肤刺激性试验和皮肤过敏试验。结果说明多拉菌素浇泼剂对家兔无任何毒性反应,对豚鼠皮肤无刺激性反应,不产生致敏作用。多拉菌素浇泼剂是一种较安全的局部外用药,适合在临床上推广应用。     

苦豆子总碱涂膜剂安全性试验研究

为研究苦豆子总碱涂膜剂在预防和治疗奶牛乳房炎方面的安全性,选用皮肤完好的健康家兔,观察家兔对苦豆子总碱涂膜剂皮肤刺激反应症状,并根据皮肤刺激反应的评分标准进行评分;选用皮肤完好的健康豚鼠进行试验,观察豚鼠对苦豆子总碱皮肤变态反应症状,根据皮肤刺激反应的评分标准进行评分。结果表明:苦豆子总碱涂膜剂皮肤刺激反应积分均值最大值为1.33,属于轻刺激性。根据皮肤变态反应的评分标准,计算所得受试物试验组去除激发受试物后24 h和48 h时的致敏率分别为25%和8.3%,属于轻度致敏强度。试验结果提示,苦豆子总碱涂膜剂临床用药安全,此结论为苦豆子总碱用于奶牛乳房炎的预防与治疗提供了依据。     

PPD的批次与注射方式对PPD皮内变态反应检疫的影响

为了探究在牛结核病检疫中出现的使用不同批次的PPD和皮下注射PPD等情况是否会对检疫结果的判定造成干扰。本试验首次通过在豚鼠背部采用皮下注射法和皮内交替注射同一厂家不同批次的结核菌素提纯蛋白衍化物(Purified protein derivative of tuberculin,PPD)的方法,参照《结核菌素提纯蛋白衍化物(PPD)制造及检定规程》于每次注射后24h、48h和72h观察皮试处是否产生迟发型变态反应。试验结果为皮内组(不同批次PPD皮内注射)和皮下组(同一批次PPD皮下注射)经3次注射后的皮试结果与标准参照组的皮试结果无差异,且在每次皮试后的24h、48h和72h皮试处均未出现局部红斑硬结。本试验证明了PPD皮内变态反应检疫结果判定不受PPD皮下接种及PPD的不同批次的干扰。     

猪瘟疫苗的豚鼠变态反应试验

本研究用猪瘟疫苗初次致敏豚鼠未引起异常反应,两周后进行第2次激发可见明显的异常反应,甚至死亡,这证明了猪瘟疫苗可能引起变态反应的发生。为了找到疫苗中的真正过敏原,本研究进行了疫苗中主要成分的分组试验,结果表明犊牛血清的致敏作用最强,其次为猪瘟病毒,牛睾丸细胞最弱。同时进行了4种致敏和激发途经的比较实验,并用ELISA法测定了致敏和激发后血清中的IgE水平。结果表明各种致敏途径均可致敏,但激发时,以静脉注射最为严重,且致敏后和激发后均未检测到血清中的IgE水平有显著提高。     

Epitheliotropic T-cell lymphoma in a guinea pig

A 4-year-old female guinea pig was presented with pruritic, erythematous, hypotrichotic and scaling skin lesions, mainly involving the abdomen and the hindlegs. Clinical abnormalities were limited to the skin, and the guinea pig was otherwise healthy. The dermatological diagnostic work-up excluded ectoparasites, dermatophytosis and an endocrinopathy as differential diagnoses for the pruritus and hair loss. An allergic dermatitis was under investigation when the general and skin condition deteriorated. A neoplastic skin disorder was suspected, and skin biopsies revealed changes consistent with a cutaneous epitheliotropic T-cell lymphoma. Pautrier's microabscesses and immunophenotyped CD3(+) CD79(-) lymphocytes were identified. Owing to the deteriorating condition of the animal and the unknown but most likely poor prognosis of this disease, the guinea pig was euthanized. To the authors' knowledge, this is the first case report of cutaneous epitheliotropic T-cell lymphoma in the guinea pig with characteristic histopathological and immunohistochemical features.     

Guinea pig protection test as indicator of potency of oil emulsion foot-and-mouth disease vaccines

A vaccine potency test is described involving virus challenge to six groups of 10 guinea pigs at five weeks after vaccination. Sixteen oil emulsion foot-and-mouth disease vaccines were so tested and nine retested after storage at 4 degrees C for up to 28.3 months. The results were compared with those of the routinely used oil emulsion vaccine potency test (protection afforded to eight pigs challenged 21 days after vaccination). When guinea pig estimates of 3 log2 PD50 or more were obtained, then, with one exception, the batches protected all or almost all pigs from challenge, but when the guinea pig estimates were less than 1 log2 PD50, the vaccines failed to protect five out of eight pigs. The sensitivity and reproducibility of the guinea pig method, established by repeated tests on two vaccine batches, seemed acceptable. The results suggested that guinea pig estimates might provide a suitable substitute for pig challenge potency tests because they reflected the potency of the vaccines, were likely to involve smaller standard errors and caused less discomfort to animals.