熊胆滴眼液药理作用的初步研究

采取常规药理实验方法研究了熊胆滴眼液的药理作用。结果显示 :熊胆滴眼液对绿脓杆菌、金黄色葡萄球菌有较强的抑菌、杀菌作用。每毫升眼药水含 2 .8× 10 3 个细菌时 ,作用 30 min有抑菌作用 ;每毫升眼药水含 2 .8× 10 2 个细菌时 ,在 5 min内能杀死绿脓杆菌 ;每毫升眼药水含2 .8× 10 2 个细菌时 ,作用 30 min有抑菌作用 ,作用 2 h能杀死金黄色葡萄球菌。熊胆滴眼液稀释1∶ 2 0以下时对 3型腺病毒感染 He L a细胞及疱疹病毒感染 L92 9细胞具有良好的保护作用 ,即有抗病毒作用 ,其对疱疹病毒作用更明显。对小鼠耳肿胀具有明显的消炎作用。熊胆滴眼液滴眼后 ,眼球结膜血流速度有所加快 ,血管管径有所增加 ,但统计学处理没有明显差异     

氯霉素眼药水使用不当导致犬角膜翳

角膜溃疡形成不透明的灰白色瘢痕称角膜翳。笔者在临床上治疗过角膜翳７２例，有７例为氯霉素使用不当引起，现介绍如下： 症 状 畏光，流泪，疼痛，结膜潮红、肿胀，眼睑闭合或半闭，角膜周围血管充血，角膜混浊或生翳膜，重者失明。 病 例 一北京犬前来就诊，主诉近来眼屎较多，遂用氯霉素眼药水滴眼，未想到反而眼疾愈发加重，眼睛开始朦朦的，后长出一层白色的东西，曾到某兽医院治疗过，用青霉素、链霉素等药物注射，未见好转。 治 疗 （１）首先用３％硼酸溶液或灭菌生理盐水冲洗患眼，清除分泌物和其他异物；（２）自家血注射：从犬…     

旋磁场治疗实验性角膜翳的效果观察

角膜翳是家畜常见的眼病之一。我们应用旋磁治疗马属动物角膜炎,获得良好效果。为了进一步验证旋磁场对角膜翳的疗效,以探讨旋磁场对治疗角膜翳有无实际应用价值,特作了本实验。一、材料和方法(一)实验动物:随机选择健康家兔20只,雌雄不拘,体重平均2.76±0.41公斤。经检查角膜及眼底均正常。     

抗菌肽对实验兔角膜损伤的疗效试验研究

为观察抗菌肽对实验兔角膜创伤的治疗效果,试验设置A组（细菌抗菌肽组）,B组（左氧氟沙星组）,C组（模型对照组）。在实验兔成功构建角膜创伤模型之后,分别使用药物进行治疗,并观察相关试验指标。试验结果显示,与模型对照组比较,细菌抗菌肽组与左氧氟沙星组实验兔的泪液分泌量均显著低于模型对照组（P<0.05）;在试验第28天的角膜荧光染色结果显示,抗菌肽组实验兔角膜的深层损伤已基本修复,其效果优于左氧氟沙星组;与模型对照组比较,抗菌肽组实验兔的眼房水中IL-1、IL-8、MMP-2等促炎因子含量水平显著降低（P<0.05）,且明显低于左氧氟沙星组;活菌计数结果显示,细菌抗菌肽对实验兔角膜创伤后眼部细菌感染的抑制作用显著高于左氧氟沙星（P<0.05）。试验结果表明,抗菌肽在治疗实验兔角膜损伤的综合疗效明显优于左氧氟沙星,为细菌抗菌肽在兽医临床上治疗宠物角膜损伤的应用提供了一定的实验数据与理论依据。     

治疗牛马角膜翳的探索

角膜翳(炎)是在牛马眼病中较常见的.临床上多是:表在性、深在性及外伤性角膜翳.本病发生后,若不及时治疗,常由急性转为慢性、白色或兰色翳膜遮盖角膜瞳孔,一眼或双眼失明.     

犬间充质干细胞因子滴眼液的制备及对角膜损伤的修复作用

制备犬间充质干细胞因子滴眼液并评价其对角膜损伤的修复效果。利用犬脂肪间充质干细胞培养上清液、氯霉素、玻璃酸钠等制备犬间充质干细胞因子滴眼液,观察温度对其性状的影响;NaOH滤纸片法制备犬角膜化学性损伤模型,评价犬间充质干细胞因子滴眼液对角膜损伤的修复作用。犬间充质干细胞因子滴眼液在4℃存放30d性状未见明显改变,滴眼液可加快角膜损伤模型角膜上皮的修复,降低眼组织综合评分。本试验制备的犬间充质干细胞因子滴眼液可促进角膜损伤的修复。辅助修复犬眼角膜因物理因素、化学因素、生物因素而造成的损伤。     

家兔角膜硷烧伤微量元素Zn、Cu含量及角膜胶原代谢变化

选体重2.5kg左右健康家兔48只,随机分为2组。实验组兔(32只)双眼角膜贴附浸有0.5mol/L NaOH液的滤纸50s后致中度碱烧伤,对照组兔(16只)不予处理。观察期间(50d),实验组兔主要呈现间歇性角膜溃疡,其角膜胶原含量和血清游离Hyp含量变化证实,角膜溃疡的发生是由于烧伤后胶原合成减少和分解加强所致;角膜Zn含量与胶原酶活性有内在联系,而Cu含量对胶原酶活性无影响。据胶原分解高峰出现时间,临床应用胶原酶抑制剂的时机宜在烧伤后7d之前。     

牛角膜混浊与翳斑的治疗

牛角膜组织受到外伤，化学刺激或结膜蔓延常引发角膜炎。由于治疗不及时常造成角膜混浊，严重的形成白色翳斑。笔者自1993年以来对患牛采用针灸太阳穴与拔云散吹眼治疗62例，取得了满意的效果。1 症状 角膜损伤部位先出现白色云雾状浑浊。随着病程的延长，流泪、羞明、敏感症状逐渐减退，但角膜不断增厚，粗糙不平。发生溃疡时角膜形成不透明的白色翳斑，边缘清晰，并常有来自角膜边缘新生血管伸入。视力常部分或大部分消失。2 治疗2．1 针灸：首先对术部和针具进行严格消毒。操作时左手紧抓牛鼻绳，右手用大拇指、食指、中指挟着圆利针后部…     

羊膜对山羊角膜缘上皮细胞培养影响的研究

比较完整羊膜和去上皮羊膜、不同冻存时间的羊膜对山羊角膜缘上皮细胞体外培养的影响,筛选出羊膜最适处理方法。将山羊角膜缘组织块分别接种于完整羊膜、去上皮羊膜、新鲜去上皮羊膜、冻存30 d去上皮羊膜、冻存90 d去上皮羊膜、冻存180 d去上皮羊膜上,比较在完整羊膜和去上皮羊膜及不同冻存时间羊膜上角膜缘上皮细胞的生长特性和组织结构。试验结果显示,在去上皮羊膜上角膜缘组织块贴壁和细胞增殖能力较完整羊膜强,角膜缘上皮细胞在2组羊膜上均能形成多层结构,但完整羊膜上形成的组织表层角质化较严重;在冻存30 d羊膜上角膜缘组织块贴壁和细胞增殖能力较新鲜羊膜、冻存90 d羊膜、冻存180 d羊膜强,角膜缘上皮细胞在各组羊膜上均形成多层结构,其结构无明显差异。结果表明,冻存30 d去上皮羊膜是构建组织工程人工角膜最适的羊膜载体材料。     

自家血注射治疗犬角膜翳

1　病因　多由于角膜外伤、异物、化学性刺激而引起 ;邻近组织病变如结膜炎、睑炎可诱发本病 ;多种传染病如犬瘟热、传染性肝炎等和某些寄生虫病时并发角膜炎 ;某些药物如氯霉素使用不当而引起 ;天气过热肝火上攻所致。2　症状　畏光 ,流泪 ,疼痛 ,结膜潮红、肿胀 ,眼睑闭合或半闭 ,角膜周围血管充血 ,角膜混浊或生翳膜 ,重者失明。3　治疗　 (1 )首先用 3%硼酸溶液或灭菌生理盐水冲洗患眼 ,清除分泌物和其他异物。(2 )自家血注射从犬臂正中静脉采血 2～ 6ml,患眼上下眼睑皮下各注射 1～ 3ml。 (3)也可用青霉素、普鲁卡因、地塞米松溶液作结…     

Duration of corneal anaesthesia following multiple doses and two concentrations of tetracaine hydrochloride eyedrops on the normal equine cornea

Reasons for performing study: There is a clinical impression that tetracaine hydrochloride (THCl) eyedrops is a suitable topical anaesthetic in horses. Objective: To determine the duration of corneal anaesthesia following instillation of multiple doses and 2 concentrations of THCl in 10 healthy horses. Methods: The corneal touch threshold (CTT) was determined, in both eyes, before (basal CTT) and after application of one drop of 0.5%THCl, 2 drops at a 1 min interval of 0.5%THCl or one drop of 1%THCl. CTT was measured in mm every 5 min until complete recovery of the basal CTT. Treatments were separated by an interval of at least one week. Results: Corneal sensitivity was significantly reduced from baseline values for 30, 60 and 50 min after application of one drop of 0.5%THCl, 2 drops of 0.5%THCl and one drop of 1%THCl, respectively. Mean maximal anaesthetic effects, corresponding to a CTT of 0 mm, lasted 5.5, 16 and 15.25 min and maximal anaesthetic effect was present in 55, 90 and 80% of eyes, 5 min after application of one drop of 0.5%THCl, 2 drops of 0.5%THCl and one drop of 1%THCl, respectively. Conclusions: The application of a second drop or the use of more concentrated eyedrops significantly increases duration of both anaesthesia and maximal anaesthetic effect. Potential relevance: Duration of corneal anaesthesia following tetracaine instillation was established enabling a better use when performing diagnostic and therapeutic procedures. Comparison of tetracaine with other ocular anaesthetics needs to be published in the future.     

siRNA抑制牛子宫内膜上皮细胞Nrf2基因的表达研究

为探讨核因子E2相关因子（nuclear factor E2 related factor，Nrf2）基因沉默及激活后对牛子宫内膜上皮细胞的影响，本试验利用小分子干扰（siRNA）技术及Nrf2的激动剂叔丁基对苯二酚（tBHQ），分别从Nrf2基因的下调和上调表达来研究Nrf2对牛子宫内膜上皮细胞中血红素加氧酶-1（HO-1）和Homebox A10（HOXA10）基因表达的影响。结果显示，经实时荧光定量PCR方法检测，在设计的2条siRNA序列（siRNA-1209和siRNA-1672）中，siRNA-1672在终浓度为75 nmol/L、作用时间24 h时抑制效果较好，抑制效率在80%以上；经Western blotting方法检测，Nrf2蛋白表达水平在转染后96 h极显著下降（P<0.01），而HO-1和HOXA10的mRNA表达量分别下降了60%和70%（P<0.01），蛋白表达量在96 h后极显著或显著下降（P<0.01；P<0.05）。此外，经CCK8方法检测，Nrf2基因表达沉默后，牛子宫内膜上皮细胞增殖能力减弱。而在tBHQ激活Nrf2试验中，经Western blotting方法检测，tBHQ终浓度为30 μmol/L时Nrf2蛋白表达量最高，极显著高于对照组（P<0.01），且HO-1和HOXA10的蛋白表达量与对照组相比也明显上升。结果表明，本试验设计的siRNA-1672能特异性地抑制牛子宫内膜上皮细胞Nrf2的表达，而抑制Nrf2的表达会导致牛子宫内膜上皮细胞增殖能力下降，且Nrf2对牛子宫内膜上皮细胞HO-1和HOXA10基因表达存在调控作用。     

阴道涂片法确定黑熊发情期的研究

本试验采用阴道涂片法对22只黑熊的阴道涂片进行了观察，结果表明，黑熊阴道上皮细胞由角化前细胞、完全角化细胞、不完全角化细胞、基底层细胞及中层细胞组成。各细胞在发情各期变化差异显著，其中，完全角化细胞在发情期数目最多，显著高于其他各期（p〈0．01）；中层细胞在发情后期显著高与其他各期（p〈0．01）；角化前细胞在发情期中差异显著（p〈0．01）。因此，通过检测各细胞的数量变化，可以确定黑熊的发情期。     

Effects of conditioned media from human amniotic epithelial cells on corneal alkali injuries in rabbits

This study was performed to evaluate the effects of conditioned media (CM) from human amniotic epithelial cells (HAECs) on the corneal wound healing process. Eighteen rabbits (36 eyes) were used and randomly assigned to three groups according treatment: CM from HAECs (group 1), vehicle alone (group 2), and saline (group 3). Corneal alkali injuries were induced with 1 N sodium hydroxide. Each reagent used for treatment evaluation was injected into the dorsal bulbar subconjunctiva and the area of the corneal epithelial defect was measured every other day. Two animals from each group were euthanized at a time on days 3, 7, and 15, and the cornea was removed for histological examination. The sum of the epithelial defect areas measured on day 0 to day 6 as well as day 0 to day 14 in group 1 was significantly smaller than those of other groups. Histological examination revealed that the group 1 corneas had less inflammatory cell infiltration and showed more intact epithelial features compared to the other groups. These results suggest that CM from HAECs promote corneal wound healing in rabbits.     

Effects of interferon-alpha on cytopathic changes and titers for feline herpesvirus-1 in primary cultures of feline corneal epithelial cells

OBJECTIVE: To assess the effect of interferon (IFN)-alpha on viability of feline corneal epithelial cells, replication of feline herpesvirus (FHV)-1, and virus-induced cytopathic changes. SAMPLE POPULATION: Healthy eyes from 10 recently euthanatized cats. PROCEDURE: 4 replicate primary cultures of feline corneal epithelial cells were grown after the addition of 10(2) to 10(6) IU of IFN-alpha/mL. Cultures were examined every 24 hours for evidence of cytotoxic changes. Viable cell counts and percentage of viable cells were determined 48 hours after initiation of culture. In a separate experiment, cultures of corneal cells were inoculated with FHV-1 and cultured for 72 hours with or without 10(5) IU of IFN-alpha/mL. The FHV-1-infected cultures were evaluated for viral-induced cytopathic effects, and viral titers were determined in samples of culture supernatant. RESULTS: Interferon-alpha did not have cytotoxic effects on corneal epithelial cells at concentrations ranging from 10(2) to 10(6) IU of IFN-alpha/mL. Interferon-alpha at a concentration of 10(5) IU/mL significantly reduced the cytopathic changes and FHV-1 titers. CONCLUSIONS AND CLINICAL RELEVANCE: Lack of in vitro cytotoxic effects and efficacy against FHV-1 infection in primary cultures of feline corneal cells suggests that the in vivo therapeutic effect of IFN-a should be assessed in controlled clinical trials.     

Effects of anti-inflammatory drugs and preservatives on morphologic characteristics and migration of canine corneal epithelial cells in tissue culture

Objective To determine the effects of commonly used ophthalmic corticosteroids, suprofen, polysulfated glycosaminoglycan and preservatives on morphologic characteristics and migration of canine corneal epithelium grown in cell culture. Animals studied Corneal epithelial cells harvested from the corneas of euthanized dogs were propagated in cell culture. Procedures Canine corneal epithelium was grown in tissue culture. The cells were treated with different corticosteroids, polysulfated glycosaminoglycan, suprofen or preservatives at different concentrations after a defect was created in the monolayer. Cellular morphologic characteristics and closure of the defect were compared between test drugs and controls. Results Morphologically the cells treated with dexamethasone were essentially the same as controls. Prednisolone and hydrocortisone caused rounding and shrinkage of the cells. Both suprofen and polysulfated glycosaminoglycan caused no apparent changes in morphologic characteristics at the lowest concentrations tested, but at higher concentrations there was a concentration‐dependent degree of rounding and shrinkage. Benzylkonium chloride and thimerosal caused rounding and shrinkage of all the cells at all concentrations tested. Dexamethasone, hydrocortisone, and suprofen did not inhibit epithelial migration over the defects at the lowest concentrations tested. All other drugs and concentrations inhibited cellular migration. Conclusion Dexamethasone affected the morphologic characteristics and migration of corneal epithelial cells less than hydrocortisone and prednisolone; therefore, dexamethasone may be the drug of choice when a corticosteroid is indicated and an epithelial defect is present. Suprofen and polysulfated glycosaminoglycan caused a concentration‐dependent effect on morphologic characteristics and migration. The preservatives caused severe changes and inhibited migration of the canine corneal epithelial cells at all concentrations and may therefore contribute to poor epithelialization of ulcers treated with preservative‐containing drugs.     

Cytopathic effects of Moraxella bovis on cultured bovine neutrophils and corneal epithelial cells

The effects of Moraxella bovis on the morphologic features of purified bovine neutrophils and bovine corneal epithelial cells were examined, using transmission and scanning electron microscopy and light microscopy. Within 2 minutes after incubation of bovine neutrophils with living M bovis, electron microscopic cellular changes included vacuolation, swelling, and loss of microplicae. Most of the neutrophils were lysed by 10 minutes of incubation. Human neutrophils phagocytosed the M bovis and remained intact, even after 30 minutes of incubation with the bacteria. Living M bovis killed bovine corneal epithelial cells in vitro. Sterile filtrates prepared from 6-hour shaker cultures of M bovis also killed bovine corneal epithelial cells, but the cytotoxic activity was less than that produced by the living bacteria. Cellular changes were first observed in specimens collected 1 hour after corneal cell monolayers were inoculated with sterile culture filtrates. The changes in these cells included pit-like lesions on the cellular surface, cellular separation, and vacuolation.     

黑白花奶牛乳腺上皮细胞冻存方法的研究

试验研究了北方地区黑白花奶牛乳腺上皮细胞的最佳冻存方法。采用内蒙古地区健康黑白花奶牛的新鲜乳腺组织,将组织进行Ⅱ型胶原酶消化后原代培养,培养的细胞经5代纯化后将密度调整为1×106个/mL。采用6种处理方法进行冻存,各设3个重复。液氮冻存1个月后,分别将6种不同方法冻存的细胞复苏后测定细胞活性,包括冻存死亡率、贴壁细胞占活细胞的比率、凋亡率等,同时进行形态学观察。结果显示,处理1组与2、3、5组间冻存死亡率存在极显著差异（P〈0.01）,处理1、4、6组间冻存死亡率无显著差异（P〉0.05）,处理2、6组间冻存死亡率存在极显著差异（P〈0.01）;处理4组的贴壁率高于处理2、5、6组,存在极显著差异（P〈0.01）。形态学观察结果发现,处理1、4组细胞较其他组生长状态良好。因此,处理4可作为黑白花奶牛乳腺上皮细胞的最佳冻存方法。     

S-亚硝基谷胱甘肽对实验性心源性肺水肿的防治作用

对小鼠腹腔注射肾上腺素造成实验性心源性肺水肿动物模型,观察S-亚硝基谷胱甘肽（GSNO）对肺水肿的防治作用。结果显示,与阳性对照相比,GSNO预防组和治疗组小鼠存活率极显著提高（P〈0.01）,肺指数极显著降低（P〈0.01）,肺组织的病理变化也明显减轻。提示：GSNO对心源性肺水肿具有明显的预防和治疗作用。