壳聚糖-胶原角膜修复材料的制备及生物相容性

背景：将胶原和壳聚糖按一定比例制成复合膜后,能够降低壳聚糖的正电荷,改善壳聚糖的吸附力,促进细胞的黏附生长、迁移和增殖,增强壳聚糖的生物学性能,成为十分优良的生物材料。 目的：制备壳聚糖-胶原复合膜,观察其与角膜基质层的组织相容性。 方法：制作壳聚糖-胶原复合膜材料。将16只新西兰兔随机分为两组,实验组于右眼角膜基质袋内植入壳聚糖-胶原复合膜,对照组右眼角膜基质袋内植入壳聚糖膜。移植后进行裂隙灯显微镜、前节-光学相干断层成像及组织学观察。 结果与结论：①裂隙灯显微镜：移植后第8周,实验组膜片中央出现降解浸润,皱褶屈曲不明显;对照组膜片全部浸润,皱褶屈曲明显。②前节-光学相干断层成像：移植后第6周,实验组的复合膜边界模糊,密度与正常角膜组织很接近,角膜形态恢复正常。③组织学观察：移植后第8周,实验组膜片表层少量降解,降解材料与角膜基质融合,膜片周围角膜基质有少量炎性细胞浸润;对照组膜片表层降解程度大于实验组,降解物质与角膜基质交织融合,膜片周围角膜基质有较多炎性细胞浸润。表明壳聚糖-胶原复合膜具有可降解性和良好的组织相容性。中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程     

Ultrasonographic findings of experimental glaucoma in rabbits

Ocular ultrasonography is a valid and non-invasive diagnostic method used to evaluate ocular and retrobulbar structures, especially when opacity of the anterior segments precludes ophthalmic examination of deeper structures of the eye or when exophthalmos is present. This study describes the B-mode ultrasonographic findings of the globe in 10 rabbits with experimental glaucoma. Ultrasonography of the eyes, using Titan TM machine and 7 MHz linear array transducer, was performed transpalpebrally. Ocular ultrasonographic findings revealed two cases of increased corneal thickness, seven cases of change in the anterior chamber depth, one case of increased echogenicity in the anterior chamber, three cases of retinal detachment, one case of increased iris and ciliary body thickness and six cases of change in axial globe length. Increased echogenicity of the capsule, cortex and nucleus was found in two cases. Six cases showed change in lens diameter. Increased echogenicity and altered lens diameter are typical of cataract. Two cases of point-like lesions, mass and/or linear echodensities in mild extent were observed within the vitreous representing haemorrhage, vitreous degeneration or detachment. The B-mode ultrasonographic imaging technique has become an essential diagnostic tool in most ocular disease. This method provides additional information allowing the clinician to offer more accurate diagnosis, treatment and prognosis in glaucoma.     

A boussinesq model of natural convection in the human eye and the formation of Krukenberg's spindle

The cornea of the human eye is cooled by the surrounding air and by evaporation of the tear film. The temperature difference between the cornea and the iris (at core body temperature) causes circulation of the aqueous humor in the anterior chamber of the eye. Others have suggested that the circulation pattern governs the shape of the Krukenberg spindle, a distinctive vertical band of pigment on the posterior cornea surface in some pathologies. We modeled aqueous humor flow the human eye, treating the humor as a Boussinesq fluid and setting the corneal temperature based on infrared surface temperature measurements. The model predicts convection currents in the anterior chamber with velocities comparable to those resulting from forced flow through the gap between the iris and lens. When paths of pigment particles are calculated based on the predicted flow field, the particles circulate throughout the anterior chamber but tend to be near the vertical centerline of the eye for a greatest period of time. Further, the particles are usually in close proximity to the cornea only when they are near the vertical centerline. We conclude that the convective flow pattern of aqueous humor is consistent with a vertical pigment spindle. © 2002 Biomedical Engineering Society. PAC2002: 4266Ew, 8710+e, 8719Pp     

Disruption of anterior segment development by TGF-beta1 overexpression in the eyes of transgenic mice.

Previous experiments showed that transgenic mice expressing a secreted self-activating transforming growth factor (TGF) -beta1 did not show a phenotype in the lens and cornea until postnatal day 21, when anterior subcapsular cataracts, sporadic thickening of the corneal stroma, and thinning of the corneal epithelium were noted (Srinivasan et al., 1998). To examine the effects of higher concentrations of TGF-beta1 on the lens and cornea, we constructed transgenic mice harboring the strong, lens-specific chicken betaB1-crystallin promoter driving an activated porcine TGF-beta1 gene. In contrast to the earlier study, the transgenic mice had microphthalmic eyes with closed eyelids. Already at embryonic day (E) 13.5, the future cornea of the transgenic mice was threefold thicker than that of wild-type littermates due to increased proliferation of corneal stromal mesenchyme cells. Staining of fibronectin and thrombospondin-1 was increased in periocular mesenchyme. At E17.5, the thickened transgenic corneal stroma was vascularized and densely populated by abundant star-shaped, neural cell adhesion molecule-positive cells of mesenchymal appearance surrounded by irregular swirls of collagen and extracellular matrix. The corneal endothelium, anterior chamber, and stroma of iris/ciliary body did not develop, and the transgenic cornea was opaque. Fibronectin, perlecan, and thrombospondin-1 were elevated, whereas type VI collagen decreased in the transgenic corneal stroma. Stromal mesenchyme cells expressed alpha-smooth muscle actin as did lens epithelial cells and cells of the retinal pigmented epithelium. By E17.5, lens fiber cells underwent apoptotic cell death that was followed by apoptosis of the entire anterior lens epithelium between E18.5 and birth. Posteriorly, the vitreous humor was essentially absent; however, the retina appeared relatively normal. Thus, excess TGF-beta1, a mitogen for embryonic corneal mesenchyme, severely disrupts corneal and lens differentiation. Our findings profoundly contrast with the mild eye phenotype observed with presumably lower levels of ectopic TGF-beta and illustrate the complexity of TGF-beta utilization and the importance of dose when assessing the effects of this growth factor.     

Pathomorphology of uveitis caused by an echovirus

The monkey eyes infested by inoculation into the eye anterior chamber of the ECHO-19 virus from children with uveitis were studied morphologically. It was established electron microscopically that the inoculated virus is replicated in the capillary endothelium and stromal cells of the uveal tract. Crystal-like accumulation of virions and degenerative changes in the organelles are observed in these cells. Acute granulomatous alterative-exudative-hemorrhagic uveitis with the destruction of the iris, ciliary body and processes and secondary changes in the cornea and crystalline lens with the symptoms of keratitis and cataract develop in the anterior part of the eye as observed in the light microscope. The anterior and posterior goniosynechia develop. An acute inflammatory process is further transformed into a progressing autoimmune inflammation.     

Autosomal dominant cataracts and Peters anomaly in a large Australian family

Peters anomaly is a congenital corneal opacity with underlying defects in the posterior stroma, Descemets membrane and corneal endothelium. It is a disorder resulting from abnormal migration or function of neural crest cells and may include abnormalities of other anterior segment structures, such as the lens and iris. We report a family in which anterior segment abnormalities, including Peters anomaly and cataracts, were inherited in an autosomal dominant fashion. Although the PAX6 gene on chromosome 11 has been shown to be involved in some cases of anterior segment developmental defects, we found no evidence that the condition in this family is linked to the PAX6 gene. Identification of this gene will indicate another gene with major involvement in the development of the anterior segment of the eye.     

A Finite Element Model for Ultrafast Laser–Lamellar Keratoplasty

A biomechanical model of the human cornea is employed in a finite element formulation for simulating the effects of Ultrafast Laser–Lamellar Keratoplasty. Several computer simulations were conducted to study curvature changes of the central corneal zone under various physiological and surgical factors. These factors included the combined effect of corneal flap and residual stromal bed thickness on corneal curvature; the effect of the shape of the lenticle on the surgical procedure outcomes and the effect of flap thickness on stress distribution in the cornea. The results were validated by comparing computed refractive power changes with clinical results. The effect of flap thickness on the amount of central flattening indicates that for flap thickness values 28% over the corneal thickness, central corneal flattening decreases. Moreover, the change in corneal curvature induced by subtraction of a plano-convex lenticle under a uniform flap, naturally imply a smaller change in the structure of the anterior layers of the cornea, but a bigger deformation in the structure of the posterior layers that are left behind the resection of the lenticle. In addition, the model also verified that the corneal curvature increased peripherally with simultaneous thinning centrally after subtraction of corneal tissue. This result shows that not only the treated zone is affected by the surgery, indicating the important role of the biomechanical response of the corneal tissue to refractive surgery, which is unaccounted for in current ablation algorithms. The results illustrate the potentialities of finite element modeling as an aid to the surgeon in evaluating variables.     

Postoperative follow up by dynamic light scattering: detection of inflammatory molecular changes in the cornea, the vitreous and the lens.

OBJECTIVE: To detect post-op molecular changes and inflammatory processes by dynamic light scattering (DLS) in eye segments. To develop DLS parameters which are appropriate for the control of post-op course after ophthalmologic surgery. METHODS, DESIGN: Three new DLS devices were tested in different post-op clinical settings such as pseudoexfoliation syndrome (PEX), cataract surgery and retinal surgery. In a second step DLS was used to detect molecular changes in the cornea of 6 patients after photo refractive keratectomy (PRK) and 16 patients after photo astigmatic refractive keratectomy (PARK). RESULTS: In PEX-patients cataract surgery changed the DLS signals of the anterior chamber and the lens in a specific way compared to the pre-op situation. Cataract patients without PEX had more scattering particles post-op and retinal surgery modified the DLS signal of the vitreous. Furthermore it was shown that the length of the corneal scattering vector l correlated with the process of the corneal wound healing after PARK. One and three months after surgery the lengths of the scattering vectors had increased significantly but returned to pre-op values twelve months post-op. DISCUSSION: The post-op transparency of eye segments such as the cornea, the vitreous and the anterior chamber depends primarily on the wound healing mechanisms. DLS seems to be a suitable method to control the corneal recovery after refractive surgery. CONCLUSIONS: The length of the scattering vector l correlates well with the wound healing after refractive surgery. However DLS-signals seem to contain information about the molecular composition of tissue too. Therefore further research is needed to refine the interpretation of the DLS signals.     

以猪角膜脱细胞基质为载体培养人脐静脉内皮细胞构建实验性角膜后板层

 目的： 探讨以异种后弹力膜/基质为载体诱导人脐静脉内皮细胞向角膜内皮细胞分化的可行性及移植术后在活体的生理功能。方法：体外分离培养脐静脉内皮细胞作为诱导移植的种子细胞;取当地质检合格的猪眼球以直径6.2 mm、厚100 μm、冷冻脱水进行角膜深板层载体的制备;接种经CM-DiI荧光标记的第2~3代人脐静脉内皮细胞于载体的后弹力层面,体外培养7~10 d行细胞形态、密度、组织学和扫描电镜观察,待细胞与后弹力层面融合形成单层后,用于兔角膜移植。受眼：正常健康新西兰大白兔24只（24眼）,实验组（人脐静脉内皮细胞移植组）12眼,对照组（单纯猪角膜深板层移植组）12眼,全角膜范围去除术眼内皮细胞,实施移植手术。结果：人脐静脉内皮细胞在猪后弹力膜/基质载体上贴附生长,形成紧密连接的单层,形态近似六角形,呈铺路石状分布,具有正常兔角膜内皮细胞的超微结构。术后8周实验组角膜基本透明,周边角膜略有水肿。对照组单纯猪角膜深板层移植后,移植角膜明显水肿、混浊。结论：以异种角膜后弹力膜/基质为载体培养的人脐静脉内皮细胞,行异种异体移植后,能够在活体上成活,并能维持角膜透明,具有正常角膜内皮细胞的生物学功能。深板层角膜内皮移植术是体外培养血管内皮细胞移植的一种有效术式。     

Keratoglobus lesions in the eyes of rearing broiler breeders.

This report describes the occurrence of keratoglobus lesions in a rearing flock of broiler breeder grandparent stock (female line), and its ophthalmological and ocular pathomorphological features. Keratoglobus is known as a recessive sex-linked (Z chromosome) trait in some lines of chickens. The first cases were encountered at 5 weeks of age. At the end of the rearing period a total of 1.5% of the flock was affected. In the flock and affected birds, no signs of any other disease were detected. Compared to eyes of healthy flock mates, the most prominent symptom was ocular protrusion due to bulging and thinning of the cornea, and increase in anterior chamber depth. In addition, there were mononuclear, granulocytic and plasma cell infiltrates in the iris and corpus ciliare, and mild to minimal exudate in the anterior chamber in all cases examined. Although the corneal epithelium and collagen bundles appeared normal on electron microscopy, the corneal endothelium showed some loss of cells and the keratocytes some degenerative changes. The cornea had not lost its transparency and did not show any signs of inflammation, whereas the conjunctiva contained mononuclear infiltrations. Other eye structures did not show any clinical or pathological changes. The pathogenesis of keratoglobus lesions in chickens is unknown. Although a hereditary origin seems probable, it is not known whether the inflammatory changes found caused the presented pathology or were secondary to the extreme corneal bulging.     

An ultrastructural investigation of dermatopontin-knockout mouse corneas

Introduction The corneal stroma is composed of a network of heterotypic collagen fibrils, proteoglycans and matrix proteins. Transparency of the tissue principally requires the uniformity of fibril diameters and interfibrillar distances and the presence of a quasi‐hexagonal lattice arrangement of parallel fibrils. Keratan sulfate proteoglycans (KSPGs) have a crucial role and the KS chains are clearly required for the maintenance of transparency. Undersulfation of corneal KS results in tissue opacity and the lumican (a KSPG) knockout mouse shows corneal opacity and the disruption of collagen fibril diameters and interfibrillar distances ( Chakravarti et al. 1998 ). Biochemical analysis has shown that dermatopontin is an abundant component of the extracellular matrix and that it interacts with KSPGs via the KS chains. This study aims to determine whether dermatopontin has a direct role in corneal matrix organization by investigating the corneal ultrastructure of dermatopontin‐null (dpt^–/–) mouse corneas. Materials and methods Conventional light microscopy was used to compare the corneal thickness of dpt^–/– mice ( Takeda et al. 2002 ) with that of the wild‐type. Collagen fibril distribution was studied using transmission electron microscopy and the datasets analysed using SIS‐pro image analysis software to determine fibrillar volume, shape factor, fibril diameter and spacing. Results Light microscopy demonstrated that dpt^–/– corneas show a 24% reduction in average stromal thickness compared to wild‐type (P < 0.001). The epithelium and Descemet's membrane appear normal. Examination of dpt^–/– stroma by transmission electron microscopy indicates a significant disruption to lamellar organization in the posterior region while the central and anterior regions appear largely unaffected compared to wild‐type. The collagen fibrils in dpt^–/– stroma show a pronounced increase in interfibrillar spacing as well as exhibiting a lower fibril volume fraction. There is no apparent difference in fibril diameter between dpt^–/– and wild‐type mice. Discussion Collectively, these data suggest that dermatopontin plays a key role in collagen fibril organization and deposition. Like the cornea from lumican‐knockout mice ( Chakravarti et al. 1998 ), the defects in collagen organization in dpt^–/– cornea appear to be most severe in the posterior stroma. It is likely that dermatopontin interacts with the KS chains on lumican and this interaction is involved in the maintenance of stromal architecture.     

多层羊膜填塞治疗微小角膜溃疡穿孔

目的观察多层羊膜填塞治疗微小角膜溃疡穿孔的临床效果,了解其有效性、安全性和局限性。方法对12例12眼角膜溃疡继发微小穿孔（穿孔直径0．5～3．0mm）患者采用多层羊膜填塞治疗。其中细菌性6例、病毒性2例、真菌性4例。清除溃疡处坏死组织,用多层羊膜填塞穿孔,其表层羊膜间断或连续缝合固定于浅层角膜上,单层羊膜再次覆盖于整个角膜表面,连续或间断缝合固定于角巩缘。术后继续使用药物治疗原发病。随访6—24个月。观察患眼溃疡愈合、前房形成、角膜厚度及并发症等。结果术后第1天,患眼前房均形成,穿孔区密闭良好,炎症控制。术后40—60d,溃疡瘢痕愈合,溃疡表面完全被角膜上皮覆盖,角膜厚度基本正常,视力均有不同程度提高,随访期间前房稳定,未出现溃疡复发及严重并发症。结论羊膜填塞治疗微小角膜溃疡穿孔安全有效。但溃疡以瘢痕愈合。     

Heightened Resistance of Athymic, Nude (nu/nu) Mice to Experimental Pseudomonas aeruginosa Ocular Infection

BALB/c-derived athymic, nude (nu/nu) mice exhibited a heightened natural resistance to experimental corneal infection with Pseudomonas aeruginosa when compared with their heterozygote (nu/+) littermates. Stereomicroscopic examination of the eyes of nu/nu mice 24 h after corneal trauma and topical bacterial application revealed slightly cloudy corneas (iris visible), whereas nu/+ littermate corneas were opaque (iris not visible). Nu/+ mice failed to resolve the infection, and endophthalmitis and shrinkage of the infected eye occurred in these mice within 2 weeks after experimental Pseudomonas infection, as in the parent BALB/c strain. However, nu/nu mice, similarly infected, resolved the infection within 24 h and never exhibited full corneal opacity or eye shrinkage. Histological examination of the corneas of nu/nu mice 24 h after experimental wounding and bacterial application demonstrated subepithelial capillaries and a few polymorphonuclear neutrophils (with numerous intracellular bacteria) in the central cornea. In contrast, the equivalent corneal areas of infected nu/+ littermates, examined similarly, showed a more striking neutrophilic response (but with few intracellular bacteria) to similar bacterial infection, as well as a lack of blood vessels within the central cornea. The central corneas of uninfected and saline control nu/nu mice also were observed. This area in nu/nu mice exhibited an infrequent polymorphonuclear neutrophil (with no intracellular bacteria) and capillaries similar in size and location to those described for experimentally infected nu/nu mouse corneas. Untreated and saline control nu/+ mice, on the other hand, lacked both vessels and polymorphonuclear neutrophils in the central cornea.     

转化生长因子β1在兔房水中的浓度变化

背景：转化生长因子β1可参与角膜损伤后的修复。 目的：观察转化生长因子β1滴眼液滴眼后房水中的浓度变化规律。 方法：将新西兰大白兔随机分为5组,分别给予PBS和质量浓度为0.5,1.0,2.0,4.0 mg/L的转化生长因子β1滴眼液滴右眼。 结果与结论：通过裂隙灯观察兔角膜和结膜结构,各组兔眼均无结膜分泌物、球结膜充血、角膜水肿增厚、角膜后沉着物、前房炎性反应及晶状体混浊改变。ELISA检测结果显示,与PBS组比较,质量浓度2.0和4.0 mg/L转化生长因子β1滴眼液能有效提高兔眼房水中转化生长因子β1的质量浓度(P < 0.01),角膜穿透性良好,在房水中可以达到有效的治疗浓度。     

兔角膜碱烧伤模型房水中肿瘤坏死因子α及血管内皮生长因子表达与姜黄素的抑制

背景：稳定的角膜新生血管动物模型是研究角膜新生血管调控机制,姜黄素对碱烧伤角膜新生血管具有抑制作用和保护作用。 目的：探讨姜黄素对碱烧伤角膜新生血管模型中肿瘤坏死因子α及血管内皮生长因子表达的影响,为防治角膜新生血管提供理论依据。 方法：纳入33只新西兰大耳白兔,随机取3只设为正常组,其余30只建立兔角膜碱烧伤诱发角膜新生血管模型,右眼设为对照组给予生理盐水,左眼设为干预组给予姜黄素,裂隙灯观察角膜新生血管生长及角膜混浊情况,酶联免疫吸附实验检测肿瘤坏死因子α和血管内皮生长因子在房水中的表达。 结果：正常组没有角膜新生血管生成。与对照组比较,干预组角膜新生血管受到抑制且角膜混浊较轻(P < 0.05)。房水肿瘤坏死因子α和血管内皮生长因子在3组中均有表达,对照组和干预组明显高于正常组,但干预组低于对照组(P < 0.05)。说明姜黄素可以有效降低角膜碱烧伤后房水肿瘤坏死因子α及血管内皮生长因子的表达进而抑制兔角膜碱烧伤后角膜新生血管的生长。     

Collagen fibril assembly during postnatal development and dysfunctional regulation in the lumican-deficient murine cornea.

The transparent cornea is the outer barrier of the eye and is its major refractive surface. Development of a functional cornea requires a postnatal maturation phase involving development, growth and organization of the stromal extracellular matrix. Lumican, a leucine-rich proteoglycan, is implicated in regulating assembly of collagen fibrils and the highly organized extracellular matrix essential for corneal transparency. We investigated the regulatory role(s) of lumican in fibril assembly during postnatal corneal development using wild type (Lum+/+) and lumican-null (Lum-/-) mice. In Lum+/+ mice, a regular architecture of small-diameter fibrils is achieved in the anterior stroma by postnatal day 10 (P10), while the posterior stroma takes longer to reach this developmental maturity. Thus, the anterior and the posterior stroma follow distinct developmental timelines and may be under different regulatory mechanisms. In Lum-/- mice, it is the posterior stroma where abnormal lateral associations of fibrils and thicker fibrils with irregular contours are evident as early as P10. In contrast, the anterior stroma is minimally perturbed by the absence of lumican. In Lum+/+ mice, lumican is expressed throughout the developing stroma at P10, with strong expression limited to the posterior stroma in the adult. Therefore, the posterior stroma, which is most vulnerable to lumican-deficiency, demonstrates an early developmental defect in fibril structure and architecture in the Lum-/- mouse. These defects underlie the reported increased light scattering and opacity detectable in the adult. Our findings emphasize the early regulation of collagen structure by lumican during postnatal development of the cornea.     

Unexpected expression of thrombospondin 1 by corneal and iris fibroblasts in the pseudoexfoliation syndrome

Pseudoexfoliation (PEX) syndrome is a common, but little known, systemic degenerative condition manifest by the extracellular deposition of a distinctive fibrillar material (PEX material) in various organs. In the eye, PEX material is characteristically found on the surfaces of structures that line the anterior and posterior chambers, and it is associated with cataract and glaucoma. Although PEX material contains several elements normally present in basement membranes, its precise composition remains obscure. Because the glycoprotein thrombospondin 1 (TSP1) can be shown in some basement membranes, we attempted to define its involvement in the composition of PEX material by immunohistochemical analysis of ocular tissues from patients with PEX syndrome. Although we were unable to detect TSP1 in PEX material, we were surprised to find that iris and corneal stromal fibroblasts expressed TSP1. In age-matched normal eyes, iris and corneal fibroblasts did not contain demonstrable TSP1. These observations indicate that TSP1 is not a significant component of PEX material but suggest that, in PEX syndrome, stromal fibroblasts remote from the principal sites of PEX material deposition are altered at the molecular level. The findings add evidence to the theory that PEX syndrome represents a disorder of connective tissue metabolism and intimate that the syndrome involves anomalous production of proteins other than those found in PEX material.     

Clinical variability of autosomal dominant cataract, microcornea and corneal opacity and novel mutation in the alpha A crystallin gene (CRYAA)

We studied 28 individuals from a four-generation Chilean family (ADC54) including 13 affected individuals with cataracts, microcornea and/or corneal opacity. All individuals underwent a complete ophthalmologic exam. We screened with a panel of polymorphic DNA markers for known loci that cause autosomal dominant cataracts, if mutated, and refined the locus using the ABI Prism Linkage Mapping Set Version 2.5, and calculated two-point lod scores. Novel PCR primers were designed for the three coding exons, including intron-exon borders, of the candidate gene alpha A crystallin (CRYAA). Clinically, affected individuals had diverse and novel cataracts with variable morphology (anterior polar, cortical, embryonal, fan-shaped, anterior subcapsular). Microcornea and corneal opacity was evident in some. Marker D21S171 gave a lod score of 4.89 (theta(m) = theta(f) = 0). CRYAA had a G414A transition that segregated with the disease and resulted in an amino acid alteration (R116H). The phenotypic variability within this family was significant with novel features of the cataracts and a corneal opacity. With the exception of iris coloboma, the clinical features in all six previously reported families with mutations in the CRYAA gene were found in this family. We identified a novel G414A transition in exon 3 of CRYAA that co-segregated with an autosomal dominant phenotype. The resulting amino acid change R116H is in a highly conserved region and represents a change in charge. The genotype-phenotype correlation of this previously unreported mutation provides evidence that other factors, genetic and/or environmental, may influence the development of cataract as a result of this alteration.     

B6-Co突变系小鼠混浊角膜组织蛋白的二维凝胶电泳分析

背景：从蛋白质组学水平研究B6-Co突变系小鼠浑浊角膜与正常B6小鼠角膜组织的异同,有利于阐释人类角膜混浊的发生机制,开辟从表型驱动研究基因功能的新途径。 目的：应用二维凝胶电泳分析技术比较分析B6和B6-Co小鼠的角膜蛋白质组学差异。 方法：分别提取B6与B6-Co突变系小鼠角膜组织蛋白,将所获得的蛋白质样品进行二维凝胶电泳和凝胶染色,分析电泳图谱,比较正常角膜组织与混浊角膜组织的蛋白质异同。 结果与结论：实验成功建立了对小鼠两种角膜组织蛋白的提取及二维凝胶电泳的基本方法和条件,二维凝胶电泳图谱清晰,通过比较分析发现B6-Co突变系小鼠混浊角膜蛋白质组中有13个蛋白质点显著下调,6个表达显著上调。B6小鼠和B6-Co突变系小鼠角膜蛋白质组有显著差异表达,提示由于基因的突变导致了相关信号通路的基因表达上调、下调或沉默。     

柔红霉素、骆驼蓬总碱对兔视网膜的毒性观察

目的 探讨药物柔红霉素(daunorubicin,DNR)、骆驼蓬总碱(total alkaloid Of Harmaline,TAH)眼内注射防治后发性白内障对视网膜的毒性作用.方法 于兔右眼晶体囊摘除(extracapsular lens extraction,ECLE)术中分别前房内注射0.2mg/mlTAH和0.2mg/ml DNR溶液0.1ml,通过眼底镜检查、视网膜电图及眼组织病理学等研究对象兔眼视网膜的影响.结果 对照眼与15只注入TAH眼眼底无明显的病理变化;TAH眼手术前后视网膜电图b波的振幅和潜时与对照眼无差别.而10只注入DNR眼出现严重的内皮性角膜水肿混浊,前房大量纤维索性渗出特等明显的炎性反应眼底及视网膜电图无法检查.组织病理学检查显示TAH组眼前节结构及视网膜各层结构正常,细胞排列规律;透射电镜检查TAH眼的视网膜感光细胞外节盘膜结构清晰.排列整齐.视网膜细胞排列紊乱,部分明显呈核固缩,表现出明显的毒性反应.结论 TAH眼内注射对兔视网膜的毒性较小,有可能用于后发性白内障的防治研究,而DNR对兔视网膜及其它组织表现出明显毒性,应进一步研究其使用的安全剂量和剂型.