MMP-2和COX-2蛋白在非小细胞肺癌组织中的表达

目的探讨基质金属蛋白酶-2（matrix metalloproteinase-2，MMP-2）和环氧合酶-2（cyclooxygenase．2，COX．2）蛋白在非小细胞肺癌组织中的表达与意义。方法应用免疫组化SP方法检测80例非小细胞肺癌组织中MMP-2，COX-2的表达。结果MMP2及COX-2的阳性率分别为31．25％、41．25％。MMP-2及COX-2蛋白表达与临床病理因素无相关性。MMP2和COX-2的阳性表达之间无相关性。结论MMP2及COX-2参与非小细胞肺癌的发生，然而它们也许并不能作为重要的预后指标。     

复方川脊片对兔颈椎病模型椎动脉基质金属蛋白酶-2的干预作用

目的:探讨基质金属蛋白酶-2(MMP-2)在椎动脉血管表达的意义及复方川脊片的干预效果。方法:40只兔随机分为正常组,模型组,颈复康组,复方川脊片组。硬化剂注射法造模,4周后,模型组,颈复康组及川脊片组分别以生理盐水、颈复康及川脊片混悬液灌胃。治疗4周后检测椎动脉血管搏动指数,全血黏度及血浆黏度,椎动脉血管MMP-2的表达。结果:与正常组相比模型组搏动指数提高,全血黏度及血浆黏度增加,MMP-2表达增多(P<0.05);与模型组相比,治疗组搏动指数降低,全血黏度及血浆黏度降低,MMP-2表达较少(P<0.05)。结论:部分的抑制MMP-2在椎动脉的表达可能是复方川脊片治疗颈椎病的机制之一。     

角膜基质细胞体外长期培养过程中生物学特性的改变

目的 研究兔角膜基质细胞在体外长期培养传代过程中,细胞增殖能力和胞外基质表达水平的改变。方法 取体外培养兔角膜基质细胞,由光镜对各代细胞的形态学变化进行观测。通过对各代细胞生长曲线的绘制,得出其各自的群体倍增时间;MTT法比较各代细胞增殖能力的改变。RT-PCR和Western杂交检测各代细胞中Ⅰ、Ⅲ型胶原表达水平的改变;爱茜蓝法检测糖胺聚糖(GAG)的含量。结果 体外培养兔角膜基质细胞随着传代的进程,逐渐显现衰老的形态特征。细胞群体倍增时间由第7代起显著延长(P<0.01);MTT比色法的结果亦显示,第7代细胞的增殖能力开始大幅下降(P<0.01)。第9-11代时,Ⅰ、Ⅲ型胶原在mRNA和蛋白水平的表达都有明显减少(P<0.01),降至第1代细胞的30％左右;GAG的含量也降至40％左右。结论 兔角膜基质细胞经历长期体外培养后,细胞增殖能力和胞外基质表达水平显著下降,逐渐丧失原有的功能。但作为构建组织工程化角膜的种子细胞,在它的生物学特性未发生改变之前,经3次传代培养,获取的细胞量达到组织块消化所得细胞的240倍左右,已足以用于构建之用。     

基质金属蛋白酶-9在腰椎间盘髓核组织中的表达

目的 检测腰椎间盘髓核组织中基质金属蛋白酶-9（MMP-9）的表达，进一步阐明椎间盘退变的机制。方法 利用RT-PCR和Western-Blot技术，对20例正常腰椎间盘髓核、45例退变腰椎间盘髓核的MMP-9进行了检测。结果 正常腰椎间盘组髓核内有一定量的MMP-9表达，退变腰椎间盘组髓核MMP-9mRNA、蛋白质表达水平升高，分别是正常组的1．86和1．65倍，存在差异（P〈0．05）。结论 MMP-9与腰椎间盘髓核退变关系密切。     

Fisher information matrix for nonlinear mixed effects multiple response models: Evaluation of the appropriateness of the first order linearization using a pharmacokinetic/pharmacodynamic model

We focus on the Fisher information matrix used for design evaluation and optimization in nonlinear mixed effects multiple response models. We evaluate the appropriateness of its expression computed by linearization as proposed for a single response model. Using a pharmacokinetic–pharmacodynamic (PKPD) example, we first compare the computation of the Fisher information matrix with approximation to one derived from the observed matrix on a large simulation using the stochastic approximation expectation–maximization algorithm (SAEM). The expression of the Fisher information matrix for multiple responses is also evaluated by comparison with the empirical information obtained through a replicated simulation study using the first‐order linearization estimation methods implemented in the NONMEM software (first‐order (FO), first‐order conditional estimate (FOCE)) and the SAEM algorithm in the MONOLIX software. The predicted errors given by the approximated information matrix are close to those given by the information matrix obtained without linearization using SAEM and to the empirical ones obtained with FOCE and SAEM. The simulation study also illustrates the accuracy of both FOCE and SAEM estimation algorithms when jointly modelling multiple responses and the major limitations of the FO method. This study highlights the appropriateness of the approximated Fisher information matrix for multiple responses, which is implemented in PFIM 3.0, an extension of the R function PFIM dedicated to design evaluation and optimization. It also emphasizes the use of this computing tool for designing population multiple response studies, as for instance in PKPD studies or in PK studies including the modelling of the PK of a drug and its active metabolite. Copyright © 2009 John Wiley & Sons, Ltd.     

Relaxin down-regulates renal fibroblast function and promotes matrix remodelling in vitro.

BACKGROUND: Renal fibroblasts are important effector cells in tubulointerstitial fibrosis, with experimental antifibrotic strategies focusing on the functional down-regulation of these cells. Several experimental models of fibrosis have provided evidence for the effectiveness of the polypeptide hormone relaxin as a potential antifibrotic agent. This study was conducted to further elucidate the antifibrotic mechanisms of relaxin on renal fibroblasts in vitro. METHODS: Rat cortical fibroblasts were obtained from outgrowth culture of renal tissue isolated from kidneys 3 days post-unilateral ureteric obstruction and constituted 100% of cells studied. A relaxin radio-receptor assay was used to establish binding of relaxin to renal fibroblasts in vitro. Functional studies then examined the effects of H2 relaxin (0, 1, 10 and 100 ng/ml) on fibroblast kinetics, expression of alpha-smooth muscle actin (alpha-SMA), total collagen synthesis, collagenase production and collagen-I lattice contraction. CTGF mRNA expression was also measured by northern analysis. RESULTS: H2 relaxin bound with high affinity to rat renal fibroblasts, but receptor numbers were low. Consistent with its previously reported bimodal effect, transforming growth factor (TGF-beta 1) reduced fibroblast proliferation, an effect abrogated by H2 relaxin. Fibroblasts exposed to H2 relaxin (100 ng/ml) for 24 h demonstrated decreased immunostaining for alpha-SMA and reduced alpha-SMA protein expression compared with controls. There was a trend for a relaxin-mediated reduction in total collagen synthesis and alpha 1(I) mRNA expression with large dose-related increases in collagenase protein expression being observed. TGF-beta 1-stimulated collagen-I lattice contraction was significantly inhibited following co-incubation with 100 ng/ml relaxin. Incremental doses of H2 relaxin had no significant effect on CTGF mRNA expression. CONCLUSIONS: The findings of this study suggest that the antifibrotic effects of relaxin involve down-regulation of fibroblast activity, increase in collagenase synthesis and restructuring of collagen-I lattices, which are consistent with its known physiological role of matrix remodelling. Although there appears to be an interaction between TGF-beta 1 and H2 relaxin, this does not appear to involve a reduction in CTGF mRNA expression.     

痔组织弹性纤维退变和血管生成的机制及其意义

目的研究痔组织弹性纤维的病理变化及微血管密度(MVD)和血管生成相关蛋白的表达与痔形成的关系。方法利用免疫组织化学染色技术(SP法),对比研究24例Ⅲ度痔患者的正常肛垫和痔两种组织内弹性纤维和MVD的变化,并检测一氧化氮合酶(NOS)的3种亚型、血管内皮生长因子(VEGF)和金属基质蛋白酶(MMP)2、MMP9在两种组织中表达的差异。结果痔组织存在显著的新生血管形成,痔组织与相对正常肛垫组织间的MVD计数、VEGF、MMP9表达差异存在统计学意义(P<0.05)。痔组织中iNOS明显增高,但与正常肛垫组织比较,不具备统计学意义;痔组织弹性纤维出现断裂、扭曲、变形、玻璃样变等异常,而正常肛垫组织中弹性纤维形态较规则、密集,断裂和变形少见。结论新生血管形成可能是痔的发病机制之一;MMP9对肛垫纤维支持结构的直接降解作用可能是痔形成和加重的一个重要机制;痔组织中iNOS表达增高还提示炎症因素和一氧化氮可能参与了痔的病理过程。     

Extracellular matrix remodeling in oral submucous fibrosis: its stage-specific modes revealed by immunohistochemistry and in situ hybridization

BACKGROUND: Oral submucous fibrosis (OSF) is a chewing habit-related pre-cancerous condition of the oral mucosa affecting predominantly south Asians. It is histopathologically characterized by epithelial atrophy and fibrosis of the subepithelial connective tissue. Fibrosis extends all the way into the muscle layer, leading to difficulty in mouth opening. However, the dynamics of extracellular matrix (ECM) remodeling with OSF progression is largely unknown. METHODS: Forty biopsy specimens of OSF and 10 of normal buccal mucosa were examined for expression/deposition modes of eight ECM molecules by histochemistry, immunohistochemistry, and in situ hybridization. RESULTS: In the early stage of OSF, tenascin, perlecan, fibronectin, collagen type III were characteristically enhanced in the lamina propria and the submucosal layer. In the intermediate stage, the ECM molecules mentioned above and elastin were extensively and irregularly deposited around muscle fibers. In the advanced stage, such ECM depositions decreased and were entirely replaced with collagen type I only. Their gene expression levels varied with progression of fibrosis, but the mRNA signals were confirmed in fibroblasts in the submucosal fibrotic areas. CONCLUSIONS: The results indicate that the ECM remodeling steps in OSF are similar to each phase of usual granulation tissue formation. Restricted mouth opening may be a result of loss of variety of ECM molecules including elastin into the homogeneity of collagen type I replacing muscle fibers.     

Stress und Grundregulation – Wirkungsweise von Akupunktur und Neuraltherapie

Stress is a general reaction to any burden on the body. If such a process becomes chronic, stress-based diseases develop which are hard to treat. The stress cycle in the hypothalamus-hypophysis-suprarenal glands disturbs the bio-psycho-social circuit, which is in a feedback loop with controlled and uncontrolled stress converting processes. This is coupled with the reactivity of the extracellular matrix (ECM) embedded in this circuit. Two “breaks” are installed in the ECM to keep stress reactions under control: The non neuronal acetylcholin system (NNAChS) and the system of endogenous endocannabinoids. Both systems have a limiting effect on pathological increased sympathetic effects raised by stress. Acupuncture stimulates the parasympathicus and NNAChS while neural therapy augments the endocannabinoid system, resulting in slower sympathetic actions.