胶原酶诱导不同部位脑出血大鼠模型的神经功能比较

目的 观察胶原酶诱导纹状体和内囊部位脑出血模型的行为学和神经纤维损伤差异.方法 利用立体定向技术,将一定量的Ⅳ型胶原酶用微量进样器分别精确注入大鼠纹状体和内囊诱导脑出血模型,观察两组大鼠的运动功能差异,并进行大体形态学和神经纤维受损程度的比较.结果 内囊组大鼠的运动功能受损程度明显重于纹状体组大鼠,前者的神经纤维破坏程度显著重于后者.结论 不同部位的脑出血模型的神经损害程度存在差异,内囊区脑出血模型更适合于研究神经纤维的损伤机制及神经纤维的再生和修复.     

胶原酶溶解术治疗腰椎间盘突出症

采用国产胶原酶行腰椎间盘内注射治疗腰椎间盘突出症９０例。有效率为９３％，优良率为８４％。作者体会该方法创伤小，无出血，对椎管内无干扰。只要严格选择适应症，掌握正确的操作方法，无疑是一种有效的治疗方法。     

介绍一种大鼠肝细胞的分离和培养方法

介绍分离、纯化大鼠肝细胞(用胶元酶二步灌流和PercoⅡ不连续密度离心)及分离后的培养。此方法可靠易行，可制备出质量合格的肝细胞(实质细胞)。讨论了保证分离和纯化成功的一些关键步骤、条件和原理．提出分离肝细胞应首选胶元酶Ⅳ，PereoⅡ的渗透压和pH也是影响肝细胞的漂浮密度和存活能力的因素。     

Histopathological and biochemical studies on pancreatic fibrosis in WBN/Kob rats

 We investigated the time-course of changes in pancreatic fibrosis accompanied with pancreatitis in WBN/Kob rats. The areas of fibrosis and fatty replacement were analysed morphometrically, and biochemical measurements of pancreatic and plasma prolyl hydroxylase and of pancreatic collagenase were assessed. Male rats showed acute pancreatitis at 2–3 months of age, lesions that later underwent a transition to widespread fibrosis. The fibrosis then decreased, and the fibrotic tissue was replaced with adipose tissue. Morphometrically, the fibrotic area reached its maximal size when the rats were 4 months old, diminishing thereafter. The fibrosis occurred mainly in the intralobular space, and was principally attributable to type-III collagen. Type-I collagen scarcely appeared throughout the experimental period. α-Smooth muscle actin appeared in and around myofibroblasts that developed in an early stage and diminished later in accordance with the progressive manner of fibrosis. The plasma prolyl hydroxylase level was higher in males than in females from 4 through 10 months of age. Pancreatic collagenase activity in the males also increased during the same period. These findings suggest that pancreatic fibrosis in male WBN/Kob rats is affected by the balance between prolyl hydroxylase and collagenase. Received: 1 October 1998 / Accepted: 2 October 1998     

Distribution of matrix metalloproteinases and their inhibitor, TIMP-1, in developing human osteophytic bone

Connective tissues synthesise and secrete a family of matrix metalloproteinases (MMPs) which are capable of degrading most components of the extracellular matrix. Animal studies suggest that the MMPs play a role in bone turnover. Using specific polyclonal antisera, immunohistochemistry was used to determine the patterns of synthesis and distribution of collagenase (MMP-1), stromelysin (MMP-3), gelatinase A (MMP-2) and gelatinase B (MMP-9) and of the tissue inhibitor of metalloproteinases-1 (TIMP-1) within developing human osteophytic bone. The different MMPs and TIMP showed distinct patterns of localisation. Collagenase expression was seen at sites of vascular invasion, in osteoblasts synthesising new matrix and in some osteoclasts at sites of resorption. Chondrocytes demonstrated variable levels of collagenase and stromelysin expression throughout the proliferative and hypertrophic regions, stromelysin showing both cell-associated and strong matrix staining. Intense gelatinase B expression was observed at sites of bone resorption in osteoclasts and mononuclear cells. Gelatinase A was only weakly expressed in the fibrocartilage adjacent to areas of endochondral ossification. There was widespread but variable expression of TIMP-1 throughout the fibrous tissue, cartilage and bone. These results indicate that MMPs play a role in the development of human bone from cartilage and fibrous tissue and are likely to have multiple functions.     

Matrix metalloproteinase expression in basal cell carcinoma: relationship between enzyme profile and collagen fragmentation pattern

Matrix metalloproteinases (MMPs) with collagenolytic and gelatinolytic activities are up-regulated in basal cell carcinoma. In the present study we demonstrate that the major collagenolytic enzyme detected is MMP-1 (interstitial collagenase) while gelatinolytic enzymes include both MMP-2 (72-kDa gelatinase A) and MMP-9 (92-kDa gelatinase B). Significant fractions of all three enzymes are present as active forms. In spite of the fact that high levels of gelatinolytic enzymes are present, the major fragmentation products resulting from digestion of intact type I collagen are the 1/4 and 3/4 fragments (products of MMP-1-mediated digestion). Thus, it appears that the gelatinolytic enzymes are not capable of degrading the collagen fragments as rapidly as they are produced. Since previous studies have demonstrated that interaction of interstitial fibroblasts with high molecular weight fragments of type I collagen leads to increased MMP production, the present results suggest a mechanism underlying altered function of stromal elements in the connective tissue adjacent to the growing neoplasm.     

鞘内注射胶原酶对兔脊髓及周围组织损伤作用的研究

目的:通过鞘内注射不同剂量的胶原酶,观察其对脊髓及周围组织的损伤作用。方法:把40只兔分成A、B、C、D、E5组,分别在鞘内注射0.3mL的生理盐水和24、50、100、250U的胶原酶,通过形态学、脊髓诱发电位和组织病理学的检查以观察胶原酶对上述组织的损伤。结果:鞘内注射胶原酶的4组兔中都有不同程度的瘫痪,没有瘫痪的兔其SCEP也有改变,病检发现脊髓、神经根和血管都有不同程度的损伤,胶原酶剂量越大,损伤越重。结论:胶原酶鞘内注射对兔脊髓及其周围组织有严重的损伤作用。     

化学髓核溶解术治疗腰椎间盘突出症

目的 探讨化学髓核溶解术的一种新的穿刺方法,用于治疗破裂型腰椎间盘突出症。方法 采用经背后入路直接穿刺椎间盘脱出部位,用600单位的胶原酶溶解脱出的椎间盘组织治疗破裂脱出型腰椎间盘突出症。本组治疗43例临床确诊为破裂脱出型腰椎间盘突出症,并随访半年至2年。结果 采用本穿刺方法治疗43例腰椎间盘脱出患者,治愈27例,治愈率为63%,有效11例,有效率为88%,无椎间盘变窄和退行性变等并发症出现。结论 经背后入路直接穿刺脱出的椎间盘组织行胶原酶溶解术是一种操作简单、安全、有效的新方法,可减少并发症的发生。     

胶原酶注射治疗腰椎间盘突出症32例随访报告

为了解胶原酶治疗腰椎间盘突出症的远期疗效,对３２例病人进行了随访。其中１５例将胶原酶４００～６００Ｕ注入病变椎间盘内（盘内注射法）;１７例将胶原酶６００～１２００Ｕ注入病变椎间盘的椎间孔附近的硬膜外间隙（盘外注射法）。３年随访结果为：优１７例、良２例、可６例、劣７例（其中４例又做开放椎间盘髓核摘除术）。优良率５９．４％,有效率７８．１％,再手术率为１２．５％,未发现明显并发症。     

The purification and partial characterisation of two novel metalloproteinases from the venom of the West African carpet viper, Echis ocellatus.

Separation of previously uncharacterised Echis ocellatus venom by phenyl-Superose FPLC (Fast Liquid Protein Chromatography) yielded eight protein fractions. Three of these displayed high proteolytic activity when assayed by in vivo and in vitro assays (including enzyme linked immunosorbant assay), and were further separated using Superdex 75 and Mono-Q FPLC. This resulted in the purification of a non-haemorrhagic 24 kDa metalloproteinase (EoVMP1, pI 7.0), and a haemorrhagic 56 kDa metalloproteinase (EoVMP2, pI 5.5). Following tryptic digest, short amino acid sequences of EoVMP1 and EoVMP2 were obtained using Edman degradation. Both sequences displayed homology when aligned with existing snake venom metalloproteinases (SVMPs). The strong homology observed among previously well-characterised SVMPs suggests that principles governing the interaction of substrates and inhibitors are likely to be similar for EoVMP1, EoVMP2 and all members of the reprolysin family.