狭鳕鱼皮胶原蛋白肽对过氧化氢诱导的人脐静脉内皮细胞氧化损伤的作用研究

目的：通过建立H2O2 导致人脐静脉内皮细胞损伤的模型来研究狭鳕鱼皮胶原蛋白肽作用。方法：用人脐静脉内皮细胞作为体外研究对象,采用不同的浓度（40M、200M、400M）胶原蛋白和维生素E预处理12h,然后在加入50uM H2O2继续培养12h造成损伤模型。模型成功后用CCK-8检测吸光度,根据试剂盒操作检测细胞内及培养上清液谷胱甘肽过氧化物酶（GXH-PX）,一氧化氮（NO）、过氧化氢酶（CAT）,丙二醛（MDA）等指标的含量和活性变化。结果：经胶原蛋白预保护后细胞增值率升高,GXH-PX、CAT等活性增强,NO含量升高,丙二醛含量降低。     

Effects of AT1 Antagonist on MMP2, MMP9 Expression and Collagen Remodeling in Left Ventricle of Rabbit Undergoing Chronic Pressure Overload

Objectives To study the effects of AT1 antagonist on MMP2, MMP9 expression and collagen remodeling in left ventricle of rabbit undergoing chronic pressure overload. Methods 30 rabbits were randomly divided into 3 groups (n= 10,each group), including sham operation group, abdominal aorta banded group(banded group), abdominal aorta banded valsartan group (valsartan group).Twelve weeks after operation,hemodynamic parame-ters were acquired, then collagen volume fraction(CVF) and MMP2, MMP9 expression of left ventricle were measured by using VG and immunohistochemical stain. Results Compared with sham operation group, both MMP2 and MMP9 expression were enhanced in banded group; meanwhile, LVW/BW,LVEDP and CVF increased significantly. Compared with banded group, both MMP2 and MMP9 expression were weakened in valarstan group; simultaneously,LVW/BW, LVEDP and CVF decreased significantly.Conclusions Expression of MMP2 and MMP9 was enhanced in left ventricle of rabbit undergoing chronic pressure overload, which may be associated with collagen proliferation, ventricule remodeling and impaired heart function; Valsartan could inhibit collagen proliferation, prevent ventricule remodeling and preserve heart function by inhibiting abnormal expression of MMP2 and MMP9.     

细胞膜表面Galectin-1抗体阻断对黏附LST-R1细胞凋亡的影响

目的观察细胞膜galectin-1抗体阻断对黏附LST-R1细胞凋亡的影响。方法激光共聚焦显微镜检测galectin-1在LST-R1细胞的表达。galectin-1抗体预孵LST-R1细胞后种植于Ⅰ型胶原（20μL/孔）包被的48孔板中,流式细胞仪检测黏附LST-R1细胞凋亡变化。结果galectin-1表达于LST-R1细胞膜表面,galectin-1抗体预孵LST-R1细胞后结合率为（67.2±6.73）%。galectin-1抗体阻断组黏附细胞呈不规则角形,而对照组黏附细胞呈圆形或类圆形;黏附LST-R1细胞凋亡率分别为（3.46±0.89）%和（8.61±1.23）%。结论LST细胞黏附至细胞外基质后,膜表面galectin-1可促进黏附细胞凋亡增加,表明膜表面galectin-1可能参与了LST病变侧向扩散非侵袭性生长特性的形成。     

Clinical usefulness of biochemical markers of liver fibrosis in patients with nonalcoholic fatty liver disease

AIM: Nonalcoholic steatohepatitis (NASH) is a severe form of nonalcoholic fatty liver disease (NAFLD), and progresses to the end stage of liver disease. Biochemical markers of liver fibrosis are strongly associated with the degree of histological liver fibrosis in patients with chronic liver disease. However, data are few on the usefulness of markers in NAFLD patients. The aim of this study was to identify better noninvasive predictors of hepatic fibrosis, with special focus on markers of liver fibrosis, type VI collagen 7S domain and hyaluronic acid. METHODS: One hundred and twelve patients with histologically proven NAFLD were studied. RESULTS: The histological stage of NAFLD correlated with several clinical and biochemical variables, the extent of hepatic fibrosis and the markers of liver fibrosis were relatively strong associated. The best cutoff values to detect NASH were assessed by using receiver operating characteristic analysis: type VI collagen 75 domain ≥5.0 ng/mL, hyaluronic acid ≥43 ng/mL. Both markers had a high positive predictive value: type VI collagen 7S domain, 86% and hyaluronic acid, 92%. Diagnostic accuracies of these markers were evaluated to detect severe fibrosis. Both markers showed high negative predictive values: type VI collagen 7S domain (≥5.0 ng/mL), 84% and hyaluronic acid (≥50 ng/mL), 78%, and were significantly and independently associated with the presence of NASH or severe fibrosis by logistic regression analysis. CONCLUSION: Both markers of liver fibrosis are useful in discriminating NASH from fatty liver alone or patients with severe fibrosis from patients with non-severe fibrosis.     

CD13/aminopeptidase N in collagen vascular diseases

To determine the significance of CD13/aminopeptidase N in collagen vascular diseases (CVD), we examined its activity and expression in sera and disease sites of patients with CVD. Significantly higher aminopeptidase activity was detected in bronchoalveolar lavage fluid from patients with interstitial lung diseases due to rheumatoid arthritis (RA), polymyositis/dermatomyositis (PM/DM), systemic sclerosis (SSc), and Sjögren's syndrome than from control subjects. Increased aminopeptidase activity and increased expression of CD13/aminopeptidase N protein were found in alveolar macrophages from CVD patients with interstitial lung diseases. Significantly higher aminopeptidase activity was detected in pleural effusions from patients with systemic lupus erythematosus (SLE) than in transudate effusions. The mean aminopeptidase activity in synovial fluids from RA patients was significantly higher than from patients with osteoarthritis. The mean value of serum aminopeptidase activity was significantly higher in patients with SLE, RA, SSc, and PM/DM than in normal subjects. This study suggests that the activity of CD13/aminopeptidase N, locally produced in the disease site, is a useful marker for CVD and that CD13/aminopeptidase N may have an important role in the pathogenesis of CVD.     

视黄醇脂对慢性酒精性肝病患者肝脏纤维化的抑制作用

目的:证明维生素A(Vit A)对慢性酒精性肝病 (ALD)患者的肝脏纤维化形成有抑制作用,为肝纤维化的治疗提供一条简便有效的新途径．方法:选择58例ALD患者,采用临床观察的方法,通过外源性的给予Vit A,以酶联免疫吸附法检查ALD患者血清Ⅲ型前胶原肽(PⅢP)及血清透明质酸(HA)的变化．结果:Vit A可以降低ALD患者血清Ⅲ型前胶原肽及血清透明质酸水平．Vit A治疗后 18 mo,血清Ⅲ型前胶原肽(16．9±6．8 μg/L vs 28．2±9．7 μg/L)及透明质酸(132．3±71 μg/L vs 210．0±87 μg／L)水平均较治疗前显著降低 (均P<0．01),且较对照组也有显著降低(PⅢP: 16．9±6．8μg/L vs 23．6±9．6μg/L,P<0．01:HA: 132．3±71 μg/L vs 192．0±97 μg／L,P<0．05)．结论:Vit A可以抑制ALD患者肝脏纤维化的形成,其机制可能是Vit A参与肝星状细胞 (HSC)的代谢,从而抑制胶原的释放．     

Steatosis recovery after treatment with a balanced sunflower or olive oil-based diet： Involvement of perisinusoidal stellate cells

AIM: To analyze the relationship between perisinusoidal stellate cell (PSC) activation and the dietary fat quantity and composition in the treatment of hepatic steatosis. METHODS: Using an experimental rat model of steatosis based on the intake of a hyperlipidic diet (14% fat as olive oil or sunflower oil, HL-O and HL-S, respectively), we analyzed the liver's capability of recovery after the treatment with a normal-lipidic diet (5% fat as olive oil or sunflower oil, NL-O and NL-S, respectively) by immu-nocytochemical and Western blot analysis of glial fibril-lary acidic protein (GFAP) expression in PSCs, collagen quantification and serum aminotransferase determination. RESULTS: The fatty infiltration in the steatotic livers decreased after the treatment with both NL diets, indicating liver recovery. This decrease was accompanied with a lower collagen deposition and aminotransferase level as well as changes in the PSC population that increased the GFAP expression. The above-mentioned effects were more pronounced in animals fed on NL-O based diet. CONCLUSION: Treatment with a balanced diet enriched in olive oil contributes to the liver recovery from a steatotic process. The PSC phenotype is a marker of this hepatic-recovery model.     

A multicenter trial of the use of the proteolytic enzyme inhibitor aprotinin in colorectal surgery

Animal studies have demonstrated the value of the proteolytic enzyme inhibitor, aprotinin, in reducing collagen breakdown and improving the healing of experimental colonic anastomoses. A double-blind, multicenter, prospective trial has evaluated the use of aprotinin in the prevention of anastomic leakage in patients. Two hundred sixteen patients undergoing colonic resection and anastomosis were studied. Patients were randomized to receive either aprotinin or placebo intravenously, peroperatively, and for the first three postoperative days. Anastomotic integrity was assessed clinically and by Hypaque^® enema on the tenth postoperative day. Although the use of aprotinin was not associated with a significant overall decrease in anastomotic leakage rates, in 95 patients undergoing anterior resection, leakage rates in those receiving aprotinin (clinical 10.8 percent; radiologic 32.4 percent) were lower than in those receiving placebo (17.2 percent and 43.1 percent, respectively). An apparent adverse association was noted, however, in patients undergoing left hemicolectomy or sigmoid colectomy who received aprotinin.     

Tracheomalacia in a Neonate with Kniest Dysplasia: Histopathologic and Ultrastructural Features

Kniest dysplasia is an autosomal-dominant chondrodysplastic condition characterized by disproportionate dwarfism, short trunk, small pelvis, kyphoscoliosis, short limbs, prominent joints, premature osteoarthritis, and craniofacial manifestations. The craniofacial abnormalities include tracheomalacia, midface hypoplasia, cleft palate, early onset myopia, retinal detachment, prominent eyes, and sensorineural hearing loss. Radiologic features include dumbbell-shaped femora, platyspondylia with anterior wedging of vertebral bodies, coronal clefts of thoracolumbar vertebral bodies, low broad ilia, and short tubular bones with broad metaphyses and deformed large epiphyses. This form of chondrodysplasia is associated with mutations in type II collagen splicing sequences. Mutations have been identified in the COL2A1 (type II collagen) gene between exons 12 and 24. Type II collagen is the predominant structural protein in cartilage, and mutations in this collagen account for the Kniest dysplasia phenotype. Histopathologic and ultrastructural features of epiphyseal plate cartilage have been described, but tracheal cartilage in an affected neonate has not been examined. The authors report the histopathologic and ultrastructural findings of anterior tracheal cartilage from a 35-day-old female with suspected chondrodysplasia who had tracheomalacia with airway obstruction. The tracheal cartilage was moderately cellular, but lacked cystic and myxoid changes in its matrix. The chondrocytes had abundant cytoplasmic PAS-positive inclusions. Some of these inclusions were diastase-resistant and were also highlighted on Alcian blue staining. Ultrastructural examination revealed chondrocytes with greatly dilated rough endoplasmic reticulum containing granular proteinaceous material. There were also frequent aggregates of typical glycogen. The defect in the COL2A1 gene is secondary to mutations, especially at splice junctions, and this markedly disrupts triple helix formation. The mutated type II procollagen results in intracellular retention within the chondrocytes, as abundant granular proteinaceous material within the dilated RER. A relationship is known to exist between the proportion of mutated to normal type II collagen in the matrix and the severity of the phenotype. With low levels of normal type II collagen, the phenotypic manifestations become more severe, such as in achondrogenesis type II. Both the quantity and quality of type II collagen modulates the phenotypic expression of type II collagenopathies.     

Rapid tissue engineering of biomimetic human corneal limbal crypts with 3D niche architecture

Limbal epithelial stem cells are responsible for the maintenance of the human corneal epithelium and these cells reside in a specialised stem cell niche. They are located at the base of limbal crypts, in a physically protected microenvironment in close proximity to a variety of neighbouring niche cells. Design and recreation of elements of various stem cell niches have allowed researchers to simplify aspects of these complex microenvironments for further study in vitro. We have developed a method to rapidly and reproducibly create bioengineered limbal crypts (BLCs) in a collagen construct using a simple one-step method. Liquid is removed from collagen hydrogels using hydrophilic porous absorbers (HPAs) that have custom moulded micro-ridges on the base. The resulting topography on the surface of the thin collagen constructs resembles the dimensions of the stromal crypts of the human limbus. Human limbal epithelial cells seeded onto the surface of the constructs populate these BLCs and form numerous layers with a high proportion of the cells lining the crypts expressing putative stem cell marker, p63α. The HPAs are produced using a moulding process that is flexible and can be adapted depending on the requirements of the end user. Creation of defined topographical features using this process could be applicable to numerous tissue-engineering applications where varied 3-dimensional niche architectures are required.