电镜观察透明质酸酶对人眼晶状体上皮细胞作用

目的：探讨透明质酸酶是否可以溶解晶状体囊膜上皮细胞及其作用机制。方法。取人眼超声乳化白内障摘除术中撕下的前囊膜，对半切开，一半立即置于37℃水浴箱中的2ml平衡液中。另一半置于37℃水浴箱中的2ml平衡液 1500IU透明质酸酶中。分别放置5、1O、15min，然后立即放入4％戊二醛中固定，送透射电镜检查。结果：置于平衡液中5、10、15min的前囊膜，囊膜表面的晶状体上皮细胞之间的细胞连接完整;而置于透明质酸酶中5，10、15min的前囊膜，其表面的晶状体上皮细胞之间以及细胞与囊膜之间的缝隙随时间延长而增大。结论:透明质酸酶可以破坏晶状体上皮细胞之间以及细胞与囊膜之间的连接，从而使晶状体囊膜上的上皮细胞易于被清除。     

人睾丸组织中成熟精子的分离方法初探

探索简单、适用的体外分离睾丸组织中成熟精子的方法,以用于男性无精子症病人行卵母细胞内单精子注射受精-胚胎移植为目的。用含不同浓度透明质酸酶的10 %人血白蛋白人类输卵管液培养人睾丸曲细精管组织匀浆2 4 h,再用Percoll梯度离心法洗涤后计算所获各级精子的总数,并设置前后对照和空白对照进行比较。用不同浓度透明质酸酶处理后的4个实验组分别与2个对照组比较,实验组所获得的各级精子的总数高于对照组,有显著差异(P<0 .0 1)。实验组中透明质酸酶浓度分别为5 0、10 0、15 0 u/ ml的10 %人血白蛋白人类输卵管液处理组获得的各级精子总数组间比较,处理前对照和处理后空白对照之间比较,无显著差异(P>0 .0 5 )。而用浓度为2 0 0 u/ ml的10 %人血白蛋白人类输卵管液处理组获得的各级精子的总数与其它浓度处理组之间比较存在统计学差异(P<0 .0 1) ,其所获得的各级精子总数相似,但活动能力较其它浓度处理组低。透明质酸酶结合Percoll梯度离心可从人睾丸曲细精管组织匀浆中分离出较多更纯的成熟精子,但过高浓度的透明质酸酶可能会影响分离出的精子的活动能力。     

玻璃酸酶预防诺维苯所致静脉炎的实验研究及临床应用

目的探讨局部外用硝酸甘油、玻璃酸酶、2%山莨菪碱、25%硫酸镁预防诺维苯(长春瑞滨,NVB)所致静脉炎的效果.方法采用NVB 0.7 mg/kg静脉推注,在静脉推注NVB前5 min分别采用上述四种药物溶液涂抹兔耳缘静脉的局部皮肤,静脉推注后于0,0.5,1,2,3 h沿静脉走向涂抹.以穿刺点及距穿刺点2 cm为中心,各切下面积为1 cm2的耳廓组织,福尔马林固定24 h做病理检查.23例患者应用NVB前后采用玻璃酸酶溶液涂抹,方法同动物实验,并与对照组相比较.结果应用不同药物涂抹后NVB对兔耳缘静脉及周围组织的损害以玻璃酸酶组最轻.临床实验组静脉炎发生率为3.9%,疼痛程度评分3.92±1.25,疼痛持续时间2.42±0.58 d;对照组静脉炎发生率为29.4%,疼痛评分6.26±1.96,疼痛持续时间8.25±2.41 d,两组差异有显著性.结论应用玻璃酸酶溶涂抹局部血管可有效地预防静脉炎的发生.     

球后阻滞麻醉中透明质酸酶对眼内压及间接眶压的影响

目的：研究球后阻滞麻醉中使用透明质酸酶对眼内压及间接眶压的影响。方法：采用双盲法，将40例40只白内障术眼随机分成两组：一组用添加150U透明质酸酶的麻药作球后阻滞麻醉，另一组则用添加生理盐水的麻药。分别测量麻醉前、麻醉后即刻、麻醉后5、10、15、20和30分钟的眼内压及间接眶压值，做对比分析。结果：球后阻滞麻醉后两组术眼的眼内压及间接眶压均较麻醉前升高，其中透明质酸酶组眼内压及间接眶压的升高幅度和持续时间均低于生理盐水组。结论：在球后阻滞麻醉中使用透明质酸酶可以降低眼内压及间接眶压的升高幅度和持续时间，增加手术安全性。     

Probing the biofunctionality of biotinylated hyaluronan and chondroitin sulfate by hyaluronidase degradation and aggrecan interaction

Molecular interactions involving glycosaminoglycans (GAGs) are important for biological processes in the extracellular matrix (ECM) and at cell surfaces, and also in biotechnological applications. Enzymes in the ECM constantly modulate the molecular structure and the amount of GAGs in our tissues. Specifically, the changeable sulfation patterns of many GAGs are expected to be important in interactions with proteins. Biotinylation is a convenient method for immobilizing molecules to surfaces. When studying interactions at the molecular, cell and tissue level, the native properties of the immobilized molecule, i.e. its biofunctionality, need to be retained upon immobilization. Here, the GAGs hyaluronan (HA) and chondroitin sulfate (CS), and synthetically sulfated derivatives of the two, were immobilized using biotin-streptavidin binding. The degree of biotinylation and the placement of biotin groups (end-on/side-on) were varied. The introduction of biotin groups could have unwanted effects on the studied molecule, but this aspect that is not always straightforward to evaluate. Hyaluronidase, an enzyme that degrades HA and CS in the ECM, was investigated as a probe to evaluate the biofunctionality of the immobilized GAGs, using both quartz crystal microbalance and high-performance liquid chromatography. Our results showed that end-on biotinylated HA was efficiently degraded by hyaluronidase, whereas already a low degree of side-on biotinylation destroyed the degrading ability of the enzyme. Synthetically introduced sulfate groups also had this effect. Hence hyaluronidase degradation is a cheap and easy way to investigate how molecular function is influenced by the introduced functional groups. Binding experiments with the proteoglycan aggrecan emphasized the influence of protein size and surface orientation of the GAGs for in-depth studies of GAG behavior.     

膈下逐瘀汤治疗乙型肝炎肝硬化临床疗效Meta分析

目的:探讨膈下逐瘀汤对乙型肝炎肝硬化疗效的Meta分析。方法:检索国内外大型文献数据库中关于膈下逐瘀汤对乙型肝炎肝硬化的随机对照实验(RCT)文献,采用Cochrane系统评价的方法,以RevMan5.1软件对数据进行分析,对所得文献进行证据质量评价。结果:获得12篇RCT文献,未提及盲法,质量普遍偏低。膈下逐瘀汤治疗乙型肝炎肝硬化的有效率较高[OR=3.25,95%CI(2.08,5.42)]。两组在乙型肝炎肝硬化HBV-DNA转阴率上,无统计学差异[OR=2.77,95%CI(0.59,13.09)]。膈下逐瘀汤治疗乙型肝炎肝硬化后Child-Pugh评分较低[OR=-1.34,95%CI(-1.61,-18.52)]。膈下逐瘀汤治疗乙型肝炎肝硬化后血ALT水平较低[OR=-27.90,95%CI(-37.28,-18.52)]。膈下逐瘀汤治疗乙型肝炎肝硬化后血HA水平低[OR=-46.72,95%CI(-62.89,-30.55)]。结论:膈下逐瘀汤可提高乙型肝炎肝硬化疗效,改善肝功能及纤维化,对HBV-DNA转阴无影响,但文献质量偏低,仍需进一步探索。     

Purification and properties of hyaluronidase from Hippasa partita (funnel web spider) venom gland extract.

Spider venom is a complex mixture of protein and peptide toxins. Hyaluronidase a 'spreading factor' has not been studied extensively in spider venom. In this paper, we describe the purification and characterization of a hyaluronidase from Hippasa partita venom gland extract. Hyaluronidase (HPHyal) has been purified by the successive chromatography on a Sephadex G-100 and on CM-Sephadex C-25 columns. HPHyal has been purified to an extent of about approximately 20-folds. The molecular mass was found to be 42.26 kDa by matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometry. HPHyal was optimally active at pH 5.8 at 37 degrees C and in the presence of 300 mM NaCl in the reaction mixture. HPHyal showed absolute specificity for hyaluronan and belongs to neutral active group of enzymes. HPHyal revealed single-precipitin line, while venom gland extract revealed multiple bands in Western blotting with the antiserum prepared against venom gland extract. HPHyal indirectly potentiates the myotoxicity of VRV-PL-VIII myotoxin and also the hemorrhagic potency of hemorrhagic complex-I. Cations, Na(+) and K(+) enhanced the activity and chloride ions do not have any effect while, divalent cations, inhibited the enzyme activity.