基于免疫共沉淀-液质联用技术的新型孕烷X受体配体检测方法的建立

目的:建立基于免疫共沉淀-液质联用技术的新型孕烷X受体(PXR)配体检测方法。方法:在HEK293细胞中转染带有Flag标签的PXR表达载体,裂解细胞后用偶联Flag抗体的微珠(beads)结合并分离细胞中表达的FlagPXR蛋白;以PXR的已知最为公认的配体/激动剂利福平为模型药物,配制1μmol/L利福平溶液,与结合有Flag-PXR蛋白的微珠孵育形成微珠-蛋白-利福平复合物;将微珠从体系中分离出来,用蛋白印迹实验检测复合物中的蛋白质,用液相色谱-质谱联用技术(液质联用技术)检测复合物中的利福平。在此基础上对利福平的作用进行验证,在肝细胞癌细胞Hep G2中检测系列浓度梯度的利福平对PXR转录因子活性的影响。结果:用免疫共沉淀技术从HEK293细胞中分离鉴定得到Flag-PXR蛋白;用液质联用技术检测到蛋白与小分子复合物中的利福平;利福平能够剂量依赖地诱导PXR的转录因子活性。结论:建立了基于免疫共沉淀-液质联用技术的新型PXR配体检测方法。     

高分子粘合剂粘鼠效果现场观察简报

运用粘合剂粘捕鼠类是一种很有效的灭鼠方法。我们试用"602”(聚甲基丙烯酸酯)的工业废料配制成粘合剂进行了现场粘鼠试验,取得较好效果。配制和使用方法:取"602"的工业废料、松香、20#机油各500克,放入容器中,用文火加热、溶解后,加饴糖150克拌和,继续加热至沸,发出饴糖香味时     

碱性染料用酸配制的原因分析

“观察植物细胞的有丝分裂”实验中介绍的染色剂有2种,一种是质量浓度为1%或2%的龙胆紫溶液(将龙胆紫溶解在质量分数为2%的醋酸溶液中配制而成的),另一种是醋酸洋红。很多教师及学生对于以上染色剂的配制有很大的疑问:碱性染料为什么用酸配制?配制好后的溶液是酸性的还是碱性的?如果是酸性的还     

利用HPLC 法测定小牛血去蛋白滴眼液中尼泊金乙酯的含量

目的:探讨高效液相色谱法(HPLC)测定小牛血去蛋白滴眼液中尼泊金乙酯含量的可行性。方法:利用Agilent 1100 Series高效液相色谱仪,在Diamonsil C18色谱柱(250 mm×4.6 mm,5μm),甲醇-0.05%乙酸水溶液(65:35)作为流动相,1.0 m L·min~(-1)流速,和30℃柱温下,利用254 nm检测波长对小牛血去蛋白滴眼液中尼泊金乙酯进行定量分析。结果:尼泊金乙酯进样量在0.29~1.43μg之间呈现良好的线性关系(r=0.9999),平均加样回收率为99.80%,RSD为0.91%。结论:HPLC方法简单快速,分离效果好,可用于小牛血去蛋白滴眼液中尼泊金乙酯的定量测定。     

抗生素利福平对Q烟粉虱防御基因表达水平的影响

通过饲喂昆虫,抗生素广泛应用于昆虫共生菌功能研究中。为了探讨抗生素对烟粉虱功能基因的直接影响,研究了通过喂食含利福平棉花叶片后烟粉虱体内乙醇脱氢酶adh-class III,knottin抗菌肽,四次跨膜蛋白tetraspanin-3,丝氨酸蛋白酶抑制剂serpin和TCP1-delta基因等5个防御基因表达量变化,结果表明50.0μg/m L利福平处理,烟粉虱adh-class III、knottin、tetraspanin-3基因表达量显著高于对照(P0.05),而serpin和TCP1-delta基因表达量没有显著差异(P0.05)。100.0μg/m L利福平处理,5个防御基因表达量均显著高于对照(P0.05)。抗生素利福平影响烟粉虱防御基因表达且与其浓度相关。本研究结果对于利用抗生素揭示昆虫共生菌功能研究具有重要参考价值。     

植物维管束透明法

1.选幼嫩蚕豆一株连根拔出,用清水洗净根、茎表面污物备用。 2.将根部浸入碱性洋红溶液(配制:取0.5克碱性洋红溶解于2毫升95%酒精中后,用自来水稀释至150毫升)中染色12小时左右,待细小叶脉染为红色后,取出,用清水漂洗数分钟。     

吸附精制破伤风类毒素质量改进试验

73年生产之吸附精制破伤风类毒素(简称吸精破类)系用钠法配制的氢氧化铝吸附剂制造的。在生产中由于颗粒大、粘度大、排气慢致使外观不理想,一度停止生产。74年曾对氨水法配制之氢氧化铝吸附剂作了一些小量吸附试验,从外观看较过去钠法吸附剂配制之吸精破类有所改善。但在75年生产中发现用氨法吸附剂配制之吸精破类PH不够稳定,经放置后PH逐步上升,有时可达9.0左右,外观也不够理想,有时也会出现颗粒大等问题。为了解决吸精破类生产当中的上述问题,我们对吸附剂加量、PH、以及精破类的纯度等因素作了一些比较试验,使吸精破类的质量得到了一定的改进,现将试验方法及所得结果报告如下:     

季节因素对过敏性结膜炎就诊及用药的影响研究

目的:探讨过敏性结膜炎(AC)与抗过敏滴眼液用量间的关系,并分析季节因素对其产生的影响。方法:采用重力沉降法于2015年全年对北京城区主要气传花粉浓度进行监测,统计2015年我院变态反应科门诊AC就诊诊次及三种主要抗过敏滴眼液处方量,并采用Spearman相关性分析AC就诊诊次与抗过敏滴眼液用量间的关系。结果:2015年全年共监测花粉76164颗,花粉分布呈现春秋两季高峰,花粉分布最高的月份为4月(34.84%),其次为3月(29.72%),5月(10.87%),9月(10.52%),8月(9.94%);花粉分布最少的月份为1月和11月(0.11%)。春季花粉高峰期(3-5月份)月平均花粉数量为19150颗,秋季花粉高峰期(8-9月份)月平均花粉数量为7792颗,秋季花粉高峰期月平均花粉数量低于春季。2015年我院变态反应科AC月平均就诊诊次为(80.42±54.28)人次,8月份AC诊次最高,占全年的18.03%,其次为9月份(16.99%)、4月份(13.99%)、3月份(12.54%)、5月份(7.98%)。抗过敏滴眼液月平均用量为(148.67±148.63)瓶,8月份抗过敏滴眼液用量最高,占全年的28.25%,其次为3月份(17.21%)、9月份(14.18%)、4月份(13.11%)、5月份(8.30%),AC就诊次数及抗过敏滴眼液处方量均呈现春秋季节高峰。Spearman相关性分析显示,每月花粉量与AC就诊诊次、抗过敏滴眼液用量呈正相关(r=0.806,0.830,P=0.000,0.000),AC就诊诊次与抗过敏滴眼液用量呈正相关(r=0.923,P=0.000)。结论:花粉分布、AC诊次及抗过敏滴眼液用量均出现春秋季节高峰。秋季花粉致敏性高于春季花粉,用药及AC就诊最高峰均出现在秋季。     

小麦苗期地下茎蛋白质双向电泳技术体系的优化

以东农冬麦1号为材料,对苗期地下茎处的蛋白提取方法、蛋白溶解、上样量、胶条的转移等方面进行了试验.结果表明:在蛋白提取方面,TCA/丙酮法(T法)和尿素/硫脲法(N法)相比T法能减少低丰度蛋白的损失得到蛋白点数更多的图谱.在蛋白溶解方面,经过两次水化液溶解的蛋纯度较高,在等电聚焦时能保持8000伏较高电压.上样量方面,10mg粗蛋白溶于两次水化液能得到清晰、分离效果好、蛋白点数较多的图像.胶条转移方面,先向胶面中加入400μl0.3%普通琼脂糖溶液后,用200μl的电极缓冲液冲洗胶条支撑膜会使胶条顺利转移到第二向胶面上且胶条与胶面间不会产生气泡.     

肺血管注射制片

材料猫、兔、豚鼠、大鼠。步骤 1.注射浆的配制 (1)取5克卡红放入40毫升蒸馏水中,边搅拌边加氨水,至完全溶解后,用滤纸过滤。 (2)取明胶20克入150毫升蒸馏水中,置水浴锅中加热充分溶解。     

珍珠明目液消炎作用观察

目的观察珍珠明目液对眼结膜刺激性及其对实验性兔眼部炎症的影响.方法给兔眼眶内滴入珍珠明目液(每次0.1ml/眼,1天3次,连续7天),观察给药7天后角膜、虹膜及结膜的反应;用30%巴豆油造成兔眼结膜炎,致炎前及致炎后局部应用珍珠明目液滴眼(0.1ml/眼),并设醋酸可的松眼药水治疗组和生理盐水对照组.于致炎后2、4、12、24、36、48及72h,根据结膜炎充血、水肿及分泌物轻重程度进行评分.结果珍珠明目液对兔眼角膜、虹膜及结膜未见明显异常反应.珍珠明目液治疗30%巴豆油所致兔眼炎症评分值显著低于相应生理盐水组(P＜0.01),与醋酸可的松治疗组比较无显著性差异.结论珍珠明目液无眼刺激性,对巴豆油致兔眼结膜炎具有较好的消炎作用.     

杞菊地黄汤联合玻璃酸钠滴眼液对肝肾阴虚型干眼症患者眼表功能和泪液MMP-2、MMP-9水平的影响

摘要 目的：观察杞菊地黄汤联合玻璃酸钠滴眼液治疗肝肾阴虚型干眼症的疗效及对眼表功能和泪液基质金属蛋白酶-2（MMP-2）、基质金属蛋白酶-9（MMP-9）的影响。方法：病例来源于2018年6月~2021年3月期间就诊于湖南省直中医医院眼科门诊的干眼症患者,共98例。采用随机数字表法分为对照组和实验组,两组均49例。对照组给予玻璃酸钠滴眼液治疗,实验组在对照组基础上接受杞菊地黄汤治疗,两组均治疗4周。对比两组疗效、中医证候积分、眼表功能和泪液MMP-2、MMP-9水平变化,观察两组治疗期间不良反应发生率。结果：实验组的临床总有效率高于对照组（P<0.05）。实验组治疗4周后眼部干涩不爽、畏光、双目频眨、口干少津、白眼泛红、舌红、苔薄、脉细证候积分均低于对照组（P<0.05）。实验组治疗4周后角膜荧光素染色（FL）评分低于对照组,泪膜破裂时间（BUT）、泪液分泌试验（SIT）水平高于对照组（P<0.05）。实验组治疗4周后泪液MMP-2、MMP-9水平低于对照组（P<0.05）。两组间不良反应发生率无统计学差异（P＞0.05）。结论：杞菊地黄汤联合玻璃酸钠滴眼液治疗肝肾阴虚型干眼症,可促进患者症状缓解,改善眼表功能,降低泪液MMP-2、MMP-9水平,安全可靠。     

Sodium sulfapyridazine in new delayed-action bases for the treatment of bacterial conjunctivitis

Possible preparation of 10 per cent sodium sulfapyridazine ophthalmic drops containing aubazidan, a microbial polysaccharide, providing prolongation of the drops action and stability on the storage was studied. The pharmacokinetic studies showed that aubazidan which is a natural polymer provided high prolongation of the sulfapyridazine effect in the tissues of the anterior part of the eye in rabbits. The prolonged effect was similar to the previously observed effect of the solutions containing synthetic polymers such as 1 per cent polyacrylamide and polyvinyl . Satisfactory tolerance by the rabbit eye tissues of 6-fold daily instillations of the 10 per cent sodium sulfapyridazine solution with aubazidan for the observation period of 1 month was stated. When stored in vials the 10 per cent sodium sulfapyridazine ophthalmic drops with 0.5 per cent of aubazidan preserved the stability for 24 months with respect to the colour, transparency, viscosity, pH and drug content. It was demonstrated with using the agar diffusion method and Staphylococcus aureus, Proteus vulgaris, Escherichia coli and Pseudomonas aeruginosa as the test cultures that sodium sulfapyridazine completely preserved its antimicrobial activity in the presence of aubazidan. The data suggested that clinical trials of the 10 per cent sodium sulfapyridazine solution with 0.2-0.5 per cent of aubazidan were promising in prevention and therapy of bacterial conjunctivitis.     

Biodegradable Ocular Inserts for Sustained Delivery of Brimonidine Tartarate: Preparation and <Emphasis Type="Italic">In Vitro</Emphasis>/<Emphasis Type="Italic">In Vivo</Emphasis> Evaluation

The bioavailability of therapeutic agents from eye drops is usually limited due to corneal barrier functions and effective eye protective mechanisms. Therefore, the current study aims to enhance ocular bioavailability of brimonidine, a potent antiglaucoma drug, through the preparation of ocular inserts. Solvent casting technique was employed to prepare the inserts using polyvinylpyrrolidone K-90 (PVP K-90) as film-forming polymer blended with different viscosity grades of bioadhesive polymers namely hydroxypropyl methycellulose, carbopol, sodium alginate, and chitosan. The prepared ocular inserts were evaluated for various physicochemical parameters, swelling behavior, and in vitro release patterns. Sodium alginate-based ocular inserts revealed the most sustainment in drug release (99% at 6 h), so it was selected for further modifications via coating it, on one side or dual sides, using hydrophobic film composed of either ethylcellulose or Eudragit RSPO. The obtained in vitro release results for the modified ocular inserts revealed that ethylcellulose is superior to Eudragit RSPO in terms of brimonidine release sustainment effect. Ocular inserts composed of 7% PVP K-90, 1.5% low molecular weight sodium alginate with or without ethylcellulose coat were able to sustain the in vitro release of brimonidine. Their therapeutic efficacy regarding intraocular pressure (IOP) lowering effect when inserted in albino rabbits eyes showed superior sustainment effect compared with that of brimonidine solution. Furthermore, due to both the mucoadhesive property and the drug sustainment effect, the one-side-coated ocular insert showed more IOP lowering effect compared with that of its non-coated or dual-side-coated counterpart.     

The use of HET-CAM test in detecting the ocular irritation

Agrochemicals must undergo numerous toxicological tests before registration. One of these experiments is the examination of eye irritation potential. To get knowledge about eye irritation, recently only the in vivo Draize-test is accepted, which is one of the most criticized methods because of the injuries inflicted on the test animals. Several in vitro methods have been used to investigate the toxicity of potential eye irritants with a view to replacing in vivo eye irritation testing. In the HET-CAM test chemicals are placed in direct contact with chorioallantoic membrane of the hen's egg. The occurrence of vascular injury or coagulation in response to a compound is the basis for employing this technique as an indication of the likelihood that a chemical would damage mucous membranes (especially the eye) in vivo. In our studies comparative screening was performed with a set of agrochemicals to establish paralell data on in vitro (HET-CAM) and in vivo (Draize) results in case of 6 agrochemicals. The solutions to be tested are added to the membrane and left in contact for 5 minutes and the membrane is examined for vascular damage at set time periods. Irritancy is scored according to the severity and speed at which damage occurs providing an indication of the likely irritant effect of the compound. Our study showed good correlation between results obtained by the HET-CAM test and those of the Draize rabbit eye test most cases. The present form of the HET-CAM test can be proposed as a pre-screen method of eye irritation tests.     

Using in vitro prediction models instead of the rabbit eye irritation test to classify and label new chemicals: a post hoc data analysis of the international EC/HO validation study

The international validation study on alternative methods to replace the Draize rabbit eye irritation test, funded by the European Commission (EC) and the British Home Office (HO), took place during 1992-1994, and the results were published in 1995. The results of this EC/HO study are analysed by employing discriminant analysis, taking into account the classification of the in vivo data into eye irritation classes A (risk of serious damage to eyes), B (irritating to eyes) and NI (non-irritant). A data set for 59 test items was analysed, together with three subsets: surfactants, water-soluble chemicals, and water-insoluble chemicals. The new statistical methods of feature selection and estimation of the discriminant functions classification error were used. Normal distributed random numbers were added to the mean values of each in vitro endpoint, depending on the observed standard deviations. Thereafter, the reclassification error of the random observations was estimated by applying the fixed function of the mean values. Moreover, the leaving-one-out cross-classification method was applied to this random data set. Subsequently, random data were generated r times (for example, r = 1000) for a feature combination. Eighteen features were investigated in nine in vitro test systems to predict the effects of a chemical in the rabbit eye. 72.5% of the chemicals in the undivided sample were correctly classified when applying the in vitro endpoints lgNRU of the neutral red uptake test and lgBCOPo5 of the bovine opacity and permeability test. The accuracy increased to 80.9% when six in vitro features were used, and the sample was subdivided. The subset of surfactants was correctly classified in more than 90% of cases, which is an excellent performance.     

高浓度玻璃酸钠滴眼液与聚乙二醇滴眼液防治飞秒激光辅助LASIK术后干眼的临床疗效对比

目的:比较高浓度玻璃酸钠滴眼液与聚乙二醇滴眼液防治飞秒激光辅助LASIK术后干眼的临床效果。方法:选取2016年1月至2017年1月在我院视光学中心收治的飞秒激光辅助LASIK术后干眼患者80例并将其随机分为A、B两组,分别给予玻璃酸钠滴眼液(3 g/L)、聚乙二醇滴眼液,在用药后1周、2周、1个月进行干眼体征检查,比较患者用药前后泪液分泌试验(SIT)、泪膜破裂时间(BUT)、角膜荧光素染色(FL)的变化。结果:术后1周,所有患者BUT均较术前显著降低,FL均较术前显著升高,差异均有统计学意义(P0.05),但SIT与术前比较差异无统计学意义(P0.05)。两组患者用药后SIT数值随时间变化差异没有统计学意义(P0.05);A组和B组分别在用药后1周、2周、1个月时进行比较,SIT变化差异没有统计学意义(P0.05);而A组BUT时间、FL评分改善时间明显早于B组,差异有统计学意义(P0.05)。结论:滴用高浓度玻璃酸钠滴眼液(3 g/L)对飞秒激光辅助LASIK术后干眼患者的效果明显优于滴用聚乙二醇。     

Assessment of the eye irritating properties of chemicals by applying alternatives to the Draize rabbit eye test: the use of QSARs and in vitro tests for the classification of eye irritation

Huggins has reported on the current situation relating to the development of alternatives to the Draize eye irritation test with rabbits, and an ECVAM Working Group have reviewed the efforts needed in order to replace this animal test within the next 10 years by using the results of non-animal assessment methods. Our report reviews regulatory experience gained over the last 20 years with the EU chemicals notification procedure with respect to the assessment of eye lesions observed in Draize tests. The nature of eye lesions and their importance for classification and labelling of possible hazards to human eyes are evaluated and discussed, with a view to promoting the development of specific in vitro assays which are able to discriminate between eye damage, moderate eye irritation, and minor irritation effects which are completely reversible within a few days. Structural alerts for the prediction of eye irritation/corrosion hazards to be classified and labelled according to international classification criteria, are presented, which should be validated in accordance with internationally agreed (OECD) principles for (Q)SAR system validation. Physicochemical limit values for prediction of the absence of any eye irritation potential relevant for human health can make available a definition of the applicability domains of alternative methods developed for the replacement of the Draize eye irritation test.     

Anti-inflammatory, antioxidant and antimicrobial activity of Ophthacare brand, an herbal eye drops.

In the present study, the herbal preparation of Ophthacare brand eye drops was investigated for its anti-inflammatory, antioxidant and antimicrobial activity, using in vivo and in vitro experimental models. Ophthacare brand eye drops exhibited significant anti-inflammatory activity in turpentine liniment-induced ocular inflammation in rabbits. The preparation dose-dependently inhibited ferric chloride-induced lipid peroxidation in vitro and also showed significant antibacterial activity against Escherichia coli and Staphylococcus aureus and antifungal activity against Candida albicans. All these findings suggest that Ophthacare brand eye drops can be used in the treatment of various ophthalmic disorders.