不同哺乳动物红细胞溶血替代眼刺激实验的比较*

背景：哺乳动物红细胞溶血实验单独或组合使用可替代体内Draize兔眼实验,用于化合物眼刺激性的标识、筛查或机制研究。但目前尚不清楚不同物种间的红细胞差异是否会对实验结果造成影响。 目的：比较大鼠和五指山小型猪血红细胞用于溶血实验的物质分类结果。 方法：分别制备两种动物的红细胞悬液,进行红细胞溶血实验。分析溶血和蛋白变性两个毒性终点,得到50%溶血浓度HD50、蛋白变性指数DI及H/D比值3个参数值。 结果与结论：经22种已知分类的眼刺激物检测表明,两种动物血样对化合物的分类差别无显著性意义(P > 0.05)。Wilcoxon符号秩和检验小型猪血与大鼠血对物质分类差异无显著性意义(S=8,P=0.304 7),经Kappa一致性检验,得出小型猪血和大鼠血的一致性一般[Kappa=0.421 1,置信区间为(0.143 3,0.698 8)]。即种属差异对物质分类结果影响不大。 关键词：眼刺激;替代方法;红细胞溶血实验;大鼠;小型猪 doi:10.3969/j.issn.1673-8225.2012.12.017     

新型保存液对家兔皮肤、眼的刺激性及标本保存的实验研究

目的：对一种新型标本保存液进行初步的毒理学测试及保存效果评定,为该产品应用的安全实用性提供科学依据。方法24只新西兰大白耳兔随机平均分为生理盐水组、传统福尔马林保存液组、混合液组、新型保存液组,观察记录各组液体对新西兰大白耳兔的皮肤、眼的刺激性及肝脏标本保存效果以及各组液体的挥发率比较,期望找出更适用于保存标本和更低保存成本的一组保存液。结果新型保存液对家兔的皮肤和眼有一定的刺激性,且强于甲醛组。混合液组保存效果最好,而新型保存液优于甲醛。结论新型保存液和甲醛的1∶1混合液更适用于保存标本和降低保存液成本。     

兔眼虹膜组织力学特性的实验研究

利用我们创建的将兔眼蝉孔水密缝合后，模拟眼内前后房压强差，对虹膜整体进行加压的实验方法，对兔眼虹膜的力学特性进行了实验研究。结果表明：兔眼虹膜是典型的粘弹性物质；面积模量与前后方压强差之间基本成线性关系。实验结果可为青光眼致盲机制解释和瞳孔阻滞力的估算提供参考。     

离体眼球表面灌流系统的建立及其功能评价

目的 建立离体眼球表面灌流系统(isolated eye superfusion system,IESS),评价IESS对离体眼球正常形态和功能的维持能力.方法 在文献报道基础上,应用仿生学原理,结合具体实验条件,自行设计制作离体眼球表面灌流系统.选取8只经检测未发生损伤的离体兔眼置于IESS中,在不同时间点对离体眼球进行角膜混浊度、荧光素滞留度、角膜水肿率和病理组织学改变进行检测,从而评价IESS的功能.以上实验间隔1周进行1次,共进行3次.结果 成功设计制作了具有自主知识产权的IESS.3组评价试验中,离体眼球角膜混浊度评分分别为0.00±0.00、0.01±0.03和0.00±0.01;荧光素滞留度评分分别为0.03±0.09、0.00±0.00和0.03±0.09;角膜水肿率分别为(2.06±1.63)%、(2.83±1.93)%和(2.03±1.19)%,以上指标均在允许范围内,并未见明显的角膜病理组织学损伤.IESS工作稳定性良好,3组实验间各指标差异无统计学意义(P＞ 0.05).结论 IESS可以维持离体眼球在实验期间的正常形态,且具有较高的稳定性和可靠性,可以用于试验用离体眼球的短期保存.     

眼外肌黏附行为对眼球运动的影响

目的 研究眼外肌黏附行为对眼球运动的影响。 方法 用 3 种不同尺寸钢性球形压头对暴露在空气中不同 时间下的离体眼外肌进行压痕-分离实验,研究眼外肌的黏附行为。 结合 JKR 理论和眼球运动模型分析眼外肌与 巩膜之间黏附作用在眼球运动过程中对维持眼球平衡的贡献。 结果 离体眼外肌肌外膜的黏附力随暴露在空气 中时间的延长而减小。 除暴露 1 h 外,黏附力在其他暴露时间下不受压头尺寸的影响。 实验黏附力与 JKR 理论计 算的理论黏附力有较好的一致性。 眼球外转过程中,眼外肌与巩膜之间的黏附力和外直肌主动力随外转角度增大 而增大。 黏附力矩随外转角度呈指数型上升,并且占阻力矩、被动力矩和黏附力矩总和的比例也越来越大。 结 论 离体眼外肌肌外膜暴露在空气中时间的长短对其黏附特性有显著影响。 JKR 理论能较好描述眼外肌与巩膜 之间的黏附作用。 该黏附作用在眼球运动中参与维持眼球的平衡,对调节眼球运动有一定的贡献。 研究结果可为 斜视等眼球运动相关疾病的研究提供理论依据。     

兔眼前房压强在体连续测量方法的研究

为了解眼睛房水压强变化规律,进一步建立闭角型青光眼模型和对疾病的治疗及视功能康复提供参考,探讨一种在体连续测量正常兔眼前房压强的方法。将正常成年新西兰白兔麻醉后,先用静脉留置针(20G)自角膜缘外行前房穿刺术,然后通过套管将Millar导管植入兔眼的前房,兔眼的前房压强信号经过Powerlab系统进行滤波、放大和模数转换后,通过Chart软件进行实时显示。结果表明:用该方法测得的正常兔眼24 h前房压强的变化范围为(1.31±0.21—2.74±0.83)KPa。前房压强的分布凌晨较低(0~5点),然后逐渐升高(5~10点),到中午或下午(11—17点)时达到最高峰,晚上又逐渐下降(18~23点)。结果证明本实验在体连续测量兔眼前房压强的方法是可行的。     

一种离体皮肤经皮电阻测量装置的研制

目的 研制一种能固定离体皮肤和稳定测量经皮电阻值的离体皮肤经皮电阻测量装置.方法 离体皮肤经皮电阻测量装置由离体皮肤固定装置和经皮电阻测量仪组成,皮肤固定装置固定离体皮肤,固定装置装有电解质溶液,皮肤与内外之间的电解质溶液形成电导池,电阻测量仪通过皮肤固定装置中的内外电极,测量离体皮肤的经皮电阻值.取同一只Wistar大鼠背侧部的8块正常离体皮肤固定于离体皮肤固定装置中,测量并记录18个时间点的经皮电阻值,以上实验重复3次.结果 检测结果在正常值范围内,3次重复实验的经皮电阻值变化曲线基本一致,说明该装置能维持离体皮肤在24 h内屏障功能的稳定.结论 离体皮肤经皮电阻测量装置能满足皮肤刺激性和（或）腐蚀性试验离体皮肤替代方法的实验要求,还能用于其他有关药物的释放和透皮吸收试验的体外方法,以及测量经皮电阻值的相关方法.     

兔颈静脉内皮细胞的原位消化、体外获取及培养

目的 建立兔颈静脉内皮细胞原位消化、体外获取及培养的方法.方法 仅解剖游离单侧兔颈静脉,并保留在原位,对侧颈静脉不进行解剖游离.阻断该静脉段的两端并插管,向该静脉段内灌注Ⅰ型胶原酶进行原位消化.切取该颈静脉段,离体状态下获取兔颈静脉内皮细胞,使用EGM-2培养基培养并传代.倒置显微镜、透射电镜观察获取的兔颈静脉内皮细胞,免疫组化法检测Ⅷ因子.结果 获取的兔颈静脉内皮细胞原代培养7～10d左右可达到80％融合.光镜下细胞为短梭形或多角形,呈“鹅卵石”样排列.透射电镜可见内皮细胞特征性的Weibel-Palade小体.兔Ⅷ因子相关抗原免疫组化检测阳性.获取的兔颈静脉内皮细胞进行冻存、复苏和传代后均可以正常生长.结论 成功建立了兔颈静脉内皮细胞原位消化、体外获取及培养的方法.     

在体和离体兔组织射频介电特性的测量

作者制作了同轴线传感器，根据同轴传输线反射原理，建立起计算机控制的生物组织介电测量系统，在６０～３０００ＭＨｚ频率范围对兔器官组织进行了有效在体和离体介电测量，５０只兔器官组织的统计分析结果表明，不同兔器官组织之间介电参数的最大差异可达３０．４％，在体和离体测量得出介电常数之间未见显著性差异，但电导率有频率依赖性的显著差异，因此将人体的离体介电参数延用到实际活体场合时，必须慎重。     

兔子宫壁组织力学特性的实验研究

目的 研究兔子宫壁组织的生物力学特性,并对分娩与未分娩组兔子宫壁力学特性进行比较.方法 以14只新西兰雌性大白兔为研究对象,取其子宫壁组织为拉伸试样,分娩与未分娩分组对照,用WDW4100微机控制电子万能实验机进行拉伸实验,测定其极限强度、应力应变关系;蠕变实验和应力松弛实验测定其黏弹性特征.结果 兔子宫壁组织表现出明显的黏弹性特征,应力应变呈指数关系;分娩组的平均极限应力为0.119MPa,未分娩组的平均极限应力为0.444MPa;分娩组的蠕变和松弛实验均明显比未分娩组低.结论 分娩与未分娩的兔子宫壁组织生物力学特性有明显的差别,对临床上研究人类子宫壁组织有一定的参考价值,有助于妇产科学的发展.     

An in vitro haemolysis test as an alternative to the draize test for ocular irritation

A haemolysis test has been evaluated as an alternative to the Draize eye test (Draize et al. 1944) for ocular irritancy. Haemolysis was seen with corrosive/ severe and severe/moderate eye irritants but was negligible with non-irritant substances.Although not quite as sensitive as dog blood to irritants, human blood gave comparable results and was also more easily available in quantity. As with other in vitro tests, some false-positive results were given by the haemolysis test and also a few false-negative results. However, in all cases these were explicable given the physical and chemical properties of the materials. pH of test solutions did not seem to be a complicating factor.There was good correlation between the results of this test and published in vivo data. Although it may not yet entirely replace the in vivo model, the haemolysis test is a valuable preliminary stage in a tiered approach to eye irritancy screening.     

数控机床金属导板辅助无牙颌种植的实验研究

目的　探讨利用数控机床（computer numerical control, CNC）制造个性化下颌无牙颌金属种植导板的新方法。 方法　8具下颌无牙颌标本随机分为两组,分别运用CNC技术加工出金属导板、运用快速成型（rapid prototyping, RP）技术加工出树脂导板并测量其在体外实验中辅助置钉的精确性。 结果　CNC与RP导板在下颌骨中部水平面上的绝对偏差分别为(0.23±0.14) mm、(1.01±0.27) mm,差异有统计学意义（P=0.000）;在矢状面上分别为(0.30±0.24) mm、(0.95±0.24) mm,差异有统计学意义（P=0.000）。 结论　在体外实验中,CNC金属下颌无牙颌种植导板辅助种植精确性高于RP树脂导板。     

Ocular toxicology: the Draize eye test

PURPOSE OF REVIEW: Superficial ocular tissues are frequently exposed to damage produced by chemical compounds applied on or around the ocular surface for cosmetic, therapeutic or accidental reasons. An experimental test measuring objectively and in a reproducible way any potential damaging effect would certainly help in prospectively minimizing unwanted effects. The Draize eye test, although commonly employed to date, does not seem to be quite satisfactory in this respect. RECENT FINDINGS: The limits of the Draize eye test and the results of some modified versions of the test are analysed. In particular, the good predictivity of the so-called low-volume Draize eye test and the recent findings of studies on the in-vitro and ex-vivo alternatives to the Draize eye test are presented. SUMMARY: The Draize eye test, despite criticisms, has been used in the clinical setting for a long time and still remains the reference protocol. To date, only a combination of alternative methods, none of which is devoid of serious criticisms, seems to be able to exhaustively recognize potential irritants and avoiding for that purpose, in some cases, the use of living animals.     

无环鸟苷抗角膜病毒新途径的实验研究

目的 研究用离心法制得的角膜胶原膜载释无环鸟苷的意义。方法 胶原膜载释药物与结膜下注射法按时间把２８只白兔分四组，每组７只，左、右眼对照，对分组采集的房水用高效液相色谱分析法测定其中的药物浓度，做比较分析。结果 ０．５ｈ结膜下注射组房水中无环鸟苷显著高于胶原膜载药组（ｔ＝４．０５，Ｐ〈０．０１）；１ｈ差异无统计学意义（ｔ＝２．０７４，Ｐ〈０．０５），３和５ｈ胶原膜组房水中药物浓度均高于结膜下注射组（ｔ值分别是４．７６１和４．１９０；Ｐ〈０．０１），并可维持较长时间。结论 通过离心法制得的胶原膜给药是胶原膜复合物替代结膜下注射的较好的给药途径。     

Allergen-induced basophil activation: CD63 cell expression detected by flow cytometry in patients allergic to Dermatophagoides pteronyssinus and Lolium perenne

BACKGROUND: In this study, we determined by flow cytometry the percentage of basophils activated after in vitro stimulation by allergens and expressing the CD63 marker. The diagnostic reliability of the technique was assessed as well as its correlation with other in vitro diagnostic parameters. METHODS: Fifty-three patients suffering from asthma and/or allergic rhinitis following sensitization to Dermatophagoides pteronyssinus and 51 patients sensitized to Lolium perenne were investigated. Twenty-four atopic patients not sensitive to these allergens and 38 healthy subjects were also selected as controls. The basophil activation test determines the percentage of basophils which express CD63 as an activation marker, by means of flow cytometry, after in vitro stimulation with allergen, using double labelling with monoclonal antibody anti-CD63-PE and anti-IgE-FITC. RESULTS: No differences in basal values (non-activated control) were found between sensitized patients, atopic controls and healthy controls. On the other hand, sensitized patients showed a significantly higher percentage of activated basophils after stimulation by allergens in vitro than both control groups (P < 0.001). We found a significant correlation between skin tests and basophil activation tests (r = 0.72, P < 0.001). We also found a positive and significant correlation between basophil activation tests and histamine release tests (r = 0.80, P < 0.001), allergen-specific sulphidoleukotriene production (r = 0.7, P < 0.001) and the occurrence of serum allergen-specific IgE (r = 0.71, P < 0.001). CONCLUSION: The basophil activation test is a highly reliable technique in the diagnosis of allergy to inhalant allergens. The sensitivity of the basophil activation test was 93.3%, and its specificity 98.4%, when using a cut-off point of 15% activated basophils as positive result.     

Irritantcy Potential and Sub Acute Dermal Toxicity Study of Pistacia Lentiscus Fatty Oil as a Topical Traditional Remedy

The current study was undertaken to assess safety of Pistacia lentiscus fruits fatty oil (PLFO) as a topical traditional remedy. A primary skin and eye irritation tests were conducted with New Zealand white rabbits to determine the potential for PLFO to produce irritation from a single application. In addition, a sub acute dermal toxicity study was performed on 18 NZW rabbits to evaluate possible adverse effect following application of PLFO for 28 days. Based on the results of the current study, PLFO is classified as slightly irritating to the skin and the eye of rabbits (Primary Irritation Index (P.I.I.) = 1.037; Ocular Irritation Index (O.I.I.) = 5.33 at 1 h). In the sub-acute toxicity test, PLFO produced neither mortality nor significant differences in the body and organ weights between control group and treated rabbits. However, a reversible irritant contact dermatitis was observed in the treated areas from the end of the second week of application until the end of experiment. This local phenomenon was accompanied by a significant skin thickening (P≤0.01) since the 12^th day (ANOVA, F = 11, 07143, P = 0, 00765) which is confirmed with an inflammatory granuloma in histological study. Haematological analysis and blood chemistry values of the 2 groups showed no significant differences in any of the parameters examined. In summary, PLFO is minimally irritating to the eye and skin after a single exposure, but it may cause irritant contact dermatitis and a reversible thickening of skin after prolonged use.     

An evaluation of the accuracy of screening antisperm antibodies using the combined GAM immunobead

A simplified method of screening for antisperm antibodies has been previously described using rabbit antihuman immunoglobulin immunobeads (GAM beads). These reports have indicated a high correlation between the GAM bead and maximal individual isotype binding. However, preliminary data in our laboratory (using a 14% cut-off criterion) indicated a high frequency of samples with positive GAM bead, but negative individual isotype results. This study was conducted to evaluate more critically the use of the GAM bead for initial antisperm antibody screening. Immunobead binding tests were performed on 98 undiluted sera. The maximal binding of the individual isotype beads (IgG, IgA, or IgM) and the GAM beads were significantly correlated (r = 0.94, P = 0.0001). However, when results were categorized as positive or negative, there was a significantly lower frequency (P less than 0.05) of positive samples using the individual isotype (46.9%) than using the GAM approach (58.2%). These data support the hypothesis that, based on continuous percent binding, the GAM bead method produces results similar to individual isotype testing. However, when data are interpreted categorically, the results may differ significantly.     

内隐职业刻板印象及其信息加工机制的研究

目的 探讨内隐职业刻板印象、内隐职业偏爱及其加工机制,更好地指导大学生和在职人员的职业选择和发展.方法 内隐联想测验和纸笔版反应与不反应联想任务.结果 ①性别主效应不显著(F-0.649,P=0.424),任务主效应显著(F=1141.930,P=0.000),性别和任务的交互作用不显著(F=0.109,P=0.743...     

异丙酚复合瑞芬太尼与七氟烷复合芬太尼对斜视术后麻醉恢复的对比研究

目的 比较异丙酚复合瑞芬太尼与七氟烷复合芬太尼对全麻术后快速动眼运动恢复时间的影响.方法 60例成年患者,随机分为两组,每组30例.R组(异丙酚复合瑞芬太尼)用异丙酚复合瑞芬太尼进行诱导及麻醉维持,S组(七氟烷复合芬太尼)用异丙酚及芬太尼诱导,七氟烷维持麻醉.麻醉停药后开始每2分钟评价麻醉恢复程度.至患者可行快速动眼运动为恢复时间,麻醉恢复后开始进行眼科评价.结果 快速动眼运动平均恢复时间,R组短于S组[(14.72±5.10)min比(23.43±5.52)min,P=0.000],差异有统计学意义.术后S组恶心呕吐发生率高于R组(37%比10%,P=0.037),差异有统计学意义.结论 需要进行即时术后眼科评估的斜视手术中,异丙酚复合瑞芬太尼更有优势.