The effect of cigarette smoke on the bovine lens

Smoking has recently been identified from epidemiological studies as a possible cause of cataract but the mechanism involved is not known. Therefore, our laboratory has initiated studies aimed at elucidating these mechanisms. Whole bovine lenses were cultured to examine possible effects of cigarette smoke on amino acid uptake and protein synthesis. Cigarette smoke, filtered to remove nicotine and tar which would not reach the eye in vivo, was bubbled through culture medium. Bovine lenses were incubated in this medium in the presence of [¹⁴C]-leucine for four days. A significant decrease in uptake of [¹⁴C]-leucine and a decrease in protein synthesis were found with smoke treated lenses. This is the first demonstration of an effect of cigarette smoke on the lens. Further work is needed to determine how this metabolic upset is mediated and how it could lead to cataract.     

Suppurative corneal ulceration in Bangladesh

Suppurative keratitis is an important preventable cause of blindness, particularly in the developing world. This study analyses 142 cases of suppurative keratitis referred to Chittagong Eye Infirmary, Bangladesh. Some 53.5% of cases were bacterial and 35.9% were fungal. The five most common pathogens were: Pseudomonas sp. 24%, Streptococcus pneumoniae 17%, Aspergillus sp. 13%, Fusarium sp. 7% and Curvularia sp. 6%. Gram stain and culture results were consistent in 62.6% of cases. Previous antibiotic treatment was a significant factor for failure of culture isolation and less so for Gram stain failure. On Gram stain, 55.9% of pseudomonal cases were missed, but only 2% of fungal cases were missed. Over all, Gram stain had a sensitivity of 62% and positive predictive value of 84% for bacterial cases, and 98% and 94% for fungal cases, respectively. Fungal ulcers were typically filamentous, but an antecedent history of trauma was not common. The most frequent injury was due to rice grains, but the inoculum appeared to be introduced during eye washing with contaminated water. Pseudomonal ulcers occurred most frequently in the monsoon season, and Fusarium cases were seen only in the hot, dry season.     

二甲亚砜对人表皮的核酸和蛋白质合成的影响

用体外培养的人的伪表皮作为模型，进行药物毒理学作用的研究，观察了二甲亚砜（ＤＭＳＯ）在不同浓度和不同接触时间条件下，对人的伪表皮细胞脱氧核糖核酸（ＤＮＡ）、核糖核酸（ＲＮＡ）和蛋白质合成的影响：随着接触时间的延长，ＤＮＡ、ＲＮＡ和蛋白质合成均受抑制。低浓度条件下（１％），ＤＮＡ、ＲＮＡ和蛋白质合成增加；在１５～５０％浓度下，ＤＮＡ和蛋白质合成抑制，而ＲＮＡ合成仍增加；在高浓度条件下（７０％～１００％），ＤＮＡ、ＲＮＡ和蛋白质合成均明显抑制。     

微量地塞米松与抗病毒药物合用对抗单疱病毒作用的影响

报告在组织培养中地塞米松对单纯疱疹病毒(HSV)的生长繁殖无影响,对无环鸟苷(ACV)和环胞苷(CC)的抗病毒作用亦无干扰。在动物实验中,微量地塞米松[0.001%,常用浓度(0.1%)的百分之一]并不恶化上皮型HSV角膜炎。同时证明有效抗病毒药物的合用,微量地塞米松仍保留有良好的消炎作用,能促进上皮型和实质层型单疱角膜炎的痊愈过程。     

Morphologic characterization of osteoblast-like cell cultures isolated from newborn rat calvaria

Summary Two methods for harvesting osteoblast-like cell populations from newborn (10 days) rat calvaria were compared. The first one consisted in culturing the periosteum-free bones and then trypsinizing the cells on the bone surface. The second one involved the migration of the osteoblasts on glass fragments before trypsinization. Since the plating efficiency, the proportion of alkaline phosphatase-positive cells, the population doubling time, and the calcium deposition were more adequate, the second method was used to further characterize the behavior of the cultures. During the first week of culture, the cells featured shapes similar to those observedin vivo on the surface of periosteum-free calvaria. They formed multilayers and, in the presence of ascorbic acid, synthetized an organic matrix containing exclusively type I collagen. Later, small amounts of type III collagen appeared. The cells were embedded in the matrix and progressively acquired the morphologic phenotype of osteocyte-like cells. The matrix mineralized in the presence of β-glycerophosphate. The technique of dropinoculation (high concentration of cells in a small volume of medium) promoted the multilayer formation and the achievement of large mineralized plates (about 1 cm²) in 3 weeks of culture.     

PsD007对四株NPC细胞放射敏感性的影响

采用ＭＴＴ方法观察癌光啉结合＾６０Ｃｏγ射线放射对四株人鼻咽癌细胞体外放射增敏作用，结果表明在一定的ＰｓＤ００７浓度范围内，随ＰｓＤ００７浓度的升高，放射对细胞的杀灭作用明显增强，当浓度分别为１．２５，６．２５，１２．５，２５和５０μｇ／ｍｌ时，对四株细胞杀灭增加倍数分别为１．０１－１．０６，１．１５－１．２９，１．２８－１．５９，１．７９－３．１８，２．８６－５．５５；放射量效存活曲线按多靶单击     

动态旋转系统构建组织工程化血管模型中血管内皮细胞分泌功能监测

目的 比较静态和动态旋转系统中构建组织工程化血管模型中血管内皮细胞分泌功能。方法新型可降解材料聚羟基丁酸酯(PHB),用胶原包埋,形成多孔状PHB 胶原管形支架。分离传代、分化人脐静脉内皮细胞,接种于PHB管型支架内腔面,分别在静态、动态旋转系统中培养14 d后,测定血管内皮细胞分泌一氧化氮(NO)、前列环素(PGI2)水平。结果 在动态旋转系统构建组织工程化血管模型中血管内皮细胞分泌NO、PGI2水平显著高于静态系统,在11 d NO分别为(120.52±3.83)μmmol／L、(80.98±5.98)μmmol／L,PGI2分另9为(20.48±1.52)μg／L,(16.59±1.29)μg／L,与静态系统相比,差异有显著性(P<0.05)。结论胶原包埋PHB支架有利于细胞的黏附和生长,可作为组织工程化血管的支架材料。在动态旋转系统构建组织工程化血管模型中血管内皮细胞,具有与正常血管类似的"生理功能"。     

分离肝细胞纯化培养方法的比较研究

目的 评价一种可提高肝细胞纯度和存活率的分离培养方法。方法 以体外两步胶原酶灌流法分离肝细胞,然后将其分成两组,对照组在接种培养前不经进一步处理,试验组则在应用适宜的Percoll梯度液离心纯化之后再行培养。藉台盼蓝拒染法比较两组肝细胞的存活率,采用MTT法动态比较两组肝细胞的增殖状态,在相差显微镜下观察细胞的纯度和形态。结果 未经进一步纯化处理的猪肝细胞存活率为90％±5％,鼠肝细胞存活率为80％±5％,两者纯度均约90％;经Percoll梯度液离心纯化后,其高活力肝细胞比率均提高至98％±2％,纯度可达99％以上。从开始接种到大部分肝细胞贴壁生长,试验组比对照组肝细胞的时间有所缩短。结论 用Percoll梯度液纯化新分离肝细胞,可提高肝实质细胞的活力与纯度。     

大鼠成体心肌细胞的分离和培养

目的: 探索稳定的大鼠成体心肌细胞的分离和培养方法. 方法: 应用主动脉逆行生物酶(5 g/L胰蛋白酶及1 g/L胶原酶)灌流法分离大鼠成体心肌细胞,免疫荧光染色法鉴定分离的心肌细胞,比较有无血清培养对心肌细胞的影响. 结果: 心肌细胞的存活率为95%以上;存活细胞呈杆状,形态完整,长宽比约为4～6∶ 1;无血清培养心肌细胞只能存活1 wk,形态不发生改变;而加血清培养心肌细胞可存活2 wk以上,但超微结构发生了改变,且不同血清浓度对成纤维细胞生长具有不同的作用. 结论: 主动脉逆行生物酶(胶原酶和蛋白酶)灌流法是比较理想的心肌细胞分离方法;血清对成体心肌细胞的培养起着重要作用.     

乳猪肝细胞的分离、培养及功能检测

目的 为构建生物人工肝进行肝细胞的准备。方法 采用胶原酶半原位灌流法分离单个乳猪肝细胞,并对其活力及单层和聚集培养后的白蛋白、尿素合成功能进行检测。结果 采用本方法从每头乳猪中分离到的单个肝细胞数为(3.1±1.5)×10~(10),活性超过95％。在加入激素和生长因子的培养基中单层培养时,肝细胞功能良好,可维持2周左右。而在未加入激素和生长因子的培养中肝细胞虽能存活1周,但功能于24h后即丧失。球形聚集培养可实现肝细胞的大量培养,且生物学活性较单层培养显著提高。结论 采用胶原酶半原位灌注法所得单个乳猪肝细胞基本能满足构建生物人工肝对肝细胞数量的要求。聚集培养接种密度大,细胞生物学活性高,可用于构建生物人工肝。