兔眼表碱烧伤后复明治疗的实验

目的:评估角膜缘移植、丝裂霉素、血管抑素对眼表碱烧伤术后免疫排斥反应的抑制作用。方法:建立兔眼表碱烧伤模型,5个实验组分别行角膜移植、角膜缘移植、丝裂霉素、血管抑素及联合治疗。观察180d的角膜新生血管、混浊度、水肿度、免疫排斥反应情况并行免疫组织化学检测。结果:新生血管、角膜混浊度、角膜水肿度、角膜排斥反应指数、角膜生存率各组均具有显著性差异;免疫反应指数大于9的样本均检测到IgG型免疫复合物沉积于角膜组织中,正常角膜未见IC的沉积。结论:角膜缘移植联合丝裂霉素和血管抑素治疗眼表碱烧伤具有良好的疗效。     

血管抑素抑制兔眼表碱烧伤角膜缘移植术后的角膜血管新生

目的:检测眼表碱烧伤角膜缘移植术后血管抑素抑制角膜新生血管的作用。方法:16只新西兰大白兔双眼制作碱烧伤模型1d后,双眼行角膜缘移植术,术后左眼局部应用血管抑素治疗2周,右眼作对照;观测4周,根据新生血管侵入角膜缘内的范围、角膜混浊与水肿程度进行分级并作统计学处理;同时测量术后7、14、21及28d的眼压。结果:术后4周时,应用血管抑素的左眼的新生血管的评分为1.19±0.10,而对照组为1.63±0.72,统计学处理显示左眼的新生血管较右眼的明显减少(P<0.05),角膜混浊与水肿程度亦明显下降。各时间点各术眼的眼压均在正常范围,无统计学差异。结论:局部应用血管抑素能有效抑制眼表碱烧伤角膜缘移植术后的新生血管增生。     

带角膜缘干细胞移植治疗翼状胬肉的临床疗效分析

目的探讨带角膜缘干细胞移植治疗翼状胬肉的临床疗效;方法手术显微镜下对235例280只眼翼状胬内行切除联合带角膜缘干细胞移植术;结果回访280只眼中263只眼移植片生长良好,3月至3年未见复发;结论带角膜缘干细胞移植术治疗翼状胬肉复发率低,只有6％。     

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。     

植床四点定位法降低深板层角膜移植后散光

背景：深板层角膜移植已广泛应用于圆锥角膜的治疗,不但能够达到与穿透性角膜移植术同样的增视效果,且具有移植后排斥反应发生率低等优点。目的：探讨采用植床四点定位法,降低深板层角膜移植后散光度的效果。方法：回顾性分析行深板层角膜移植的圆锥角膜病例50例(50眼),分别在移植中采用植床四点定位法25例(25眼)和直接剖切植床即未采用植床四点定位法25例(25眼),对比观察两组患者治疗后散光度及角膜地形图模拟角膜镜度数。结果与结论：移植后随访6-12个月,平均8.3个月。两组患者无角膜植片排斥反应、继发感染、继发性青光眼等发生。采用植床四点定位法,治疗后患者散光度及角膜地形图模拟角膜镜度数均较未采用四点定位降低。深板层角膜移植术中采用植床四点定位法能够有效地减少因眼内压下降、眼球变形、植床扭曲造成的移植后散光。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程全文链接：     

自体角膜缘干细胞移植联合丝裂霉素C治疗翼状胬肉疗效观察

目的：观察自体角膜缘干细胞移植术联合丝裂霉素C应用治疗翼状胬肉的疗效。方法对102例122只眼行胬肉切除,将同侧眼颞上方带有角膜缘干细胞的结膜瓣移植到胬肉处的巩膜上,术中联合应用0.2mg/ml丝裂霉素C治疗。结果术后反应轻,移植片存活,4只眼复发,复发率为3.3%。结论自体角膜缘干细胞移植术联合丝裂霉素C应用治疗翼状胬肉,方法简单,恢复角膜缘屏障,降低了复发率,是安全、有效的方法。     

准分子激光联合板层角膜移植术在角膜瘢痕性混浊治疗中的应用

目的：探讨应用准分子激光联合板层角膜移植术治疗角膜瘢痕性混浊眼病的临床疗效。方法：对20例病变深度均未累及角膜后弹力层和内皮细胞层的角膜瘢痕性混浊患者（其中角膜云翳5例、斑翳3例、白斑12例）,按照角膜混浊的范围和累及层次,分别采用准分子激光新技术行PTK、LASIK板层或全厚植片深板层角膜移植术进行治疗;术后观察患者角膜植片愈合情况、角膜内皮细胞密度,并比较手术前后光学增视效果。结果：20例患者角膜植床和植片愈合良好,随访期内均未发生排斥现象,手术眼视力较手术前均有不同程度提高。结论：按照病变累及角膜组织解剖上的不同深度,合理选择激光联合板层角膜移植术的不同术式治疗角膜瘢痕性混浊,治疗效果良好,并发症少,免疫排斥发生率低,可在临床推广应用。     

丝裂霉素联合自体角膜移植术治疗翼状胬肉50例分析

目的探讨丝裂霉素C联合自体角膜缘干细胞移植治疗翼状胬肉的效果。方法对50例（75）眼翼状胬肉患者行翼状胬肉切除和丝裂霉素C联合自体角膜缘干细胞移植术。结果50例（75眼）翼状胬肉患者随访12-24个月，2例复发。结论翼状胬肉切除和丝裂霉素C联合自体角膜缘干细胞移植术疗效好．可较大地降低胬肉术后的复发率。     

角膜干细胞重建眼表后泪膜稳定性改变的试验研究

目的：研究角膜缘干细胞重建眼表术后泪膜生理功能改变，探讨利用角膜缘干细胞移植重建眼表的有效性和评价指标。方法：以健康雄性新西兰兔为实验对象，取左眼角膜缘干细胞体外培养，然后进行眼表重建，观察泪膜生理功能改变情况。结果：采用羊膜为载体培养角膜缘干细胞，移植修复眼表结构后，眼表细胞形态与烧伤前相似；泪膜破裂时间测试，修复后与烧伤后有显著差异（P<0.05），修复后与烧伤前无显著差异（P>0.05）。结论：利用角膜缘干细胞培养可能是重建眼表有效途径，修复后细胞结构形态和泪膜生理功能恢复良好；眼表细胞结构形态分析和泪膜破裂时间是良好的眼表重建疗效分析指标。     

雷公藤滴眼液防治大鼠角膜移植排斥反应的实验研究

目的：探讨并比较雷公藤多甙（T_Ⅱ）滴眼液及雷公藤内酯醇（T₁₀）滴眼液局部应用对角膜移植免疫排斥反应的影响。方法：建立封闭群大鼠同种异体角膜移植模型，随机分组：A、B、C组为SD-Wistar组间同种异体角膜移植，SD为受体，Wistar为供体，其中A组为空白对照组、B组为1 mg/L T₁₀滴眼液组、C组为0.03%（300 mg/L）T_Ⅱ滴眼液组，另设D组为SD-SD对照组，即SD大鼠间同种异体移植组和E组为SD大鼠自体角膜移植组。用裂隙灯显微镜记录及比较各组移植排斥指数（RI），包括：角膜透明度、水肿度、新生血管度以及角膜排斥发生时间。结果：术后各组角膜植片透明度、植片水肿度、角膜新生血管以及角膜移植排斥时间，A组与B组、A组与C组、B组与C组比较均有极显著差异（P<0.01）；D组和E组两对照组之间比较无显著差异（P>0.05）。病理学检查：B、C组15 d时的角膜植片淋巴细胞浸润明显少于A组，新生血管减少；免疫组化显示：ICAM-1和IL-2表达在15 d的B、C组角膜植片上明显少于A组。结论：T_Ⅱ滴眼液可有效地防治角膜移植免疫排斥反应，0.03% T_Ⅱ比1 mg/L T₁₀滴眼液更有效。     

Blockade of the 4-1BB (CD137)/4-1BBL and/or CD28/CD80/CD86 costimulatory pathways promotes corneal allograft survival in mice

To explore the roles of 4-1BB (CD137) and CD28 in corneal transplantation, we examined the effect of 4-1BB/4-1BB ligand (4-1BBL) and/or CD28/CD80/CD86 blockade on corneal allograft survival in mice. Allogeneic corneal transplantation was performed between two strains of wild-type (WT) mice, BALB/c and C57BL/6 (B6), and between BALB/c and B6 WT donors and various gene knockout (KO) recipients. Some of the WT graft recipients were treated intraperitoneally with agonistic anti-4-1BB or blocking anti-4-1BBL monoclonal antibody (mAb) on days 0, 2, 4 and 6 after transplantation. Transplanted eyes were observed over a 13-week period. Allogeneic grafts in control WT B6 and BALB/c mice treated with rat immunoglobulin G showed median survival times (MST) of 12 and 14 days, respectively. Allogeneic grafts in B6 WT recipients treated with anti-4-1BB mAb showed accelerated rejection, with an MST of 8 days. In contrast, allogeneic grafts in BALB/c 4-1BB/CD28 KO and B6 CD80/CD86 KO recipients had significantly prolonged graft survival times (MST, 52.5 days and 36 days, respectively). Treatment of WT recipients with anti-4-1BB mAb resulted in enhanced cellular proliferation in the mixed lymphocyte reaction and increased the numbers of CD4(+) CD8(+) T cells, and macrophages in the grafts, which correlated with decreased graft survival time, whereas transplant recipients with costimulatory receptor deletion showed longer graft survival times. These results suggest that the absence of receptors for the 4-1BB/4-1BBL and/or CD28/CD80/CD86 costimulatory pathways promotes corneal allograft survival, whereas triggering 4-1BB with an agonistic mAb enhances the rejection of corneal allografts.     

Renal Allograft Infiltrating Lymphocytes: Frequency of Tissue Specific Lymphocytes

Acute rejection, mediated by T lymphocytes recognizing donor MHC class I and II, is a major factor influencing renal transplant survival. To define the specificity of these effector cells we examined cytolytic activity of graft infiltrating T lymphocytes (GIL) from renal biopsies of individuals undergoing acute cellular rejection. The majority of these cells recognized MHC class I on both donor kidney epithelial cells (KCL) and B-lymphoblastoid cells (LCL) suggesting these T cells recognized peptides from various tissues. However, cold target inhibition experiments demonstrated a significant proportion of GIL T cells were tissue specific. We reported previously that kidney specific CTL can be isolated from biopsies of kidney allografts undergoing acute cellular rejection. Here we extend that observation showing we were able to isolate tissue specific CTL from two additional biopsies. Greater than 10% of the clones isolated (4 of 36 and 5 of 37) from these biopsies were CTL recognizing donor KCL but not LCL targets suggesting that peptides, recognized in the context of donor MHC, were tissue specific. Repeated isolation of significant numbers of tissue specific CTL suggests these T cells play a role in allograft rejection and may be important effector cells mediating rejection in HLA matched transplants.     

Percutaneous needle biopsies of renal allografts: the relationship between morphological changes present in biopsies and subsequent allograft function

In the Cambridge renal transplant unit percutaneous needle biopsies of renal transplants have been extensively used to help identify the cause of impaired allograft function. During the period 1966--1973, 154 of the 269 renal allografts transplanted were biopsied at least once during the first 90 days after transplantation. In this survey the relationship between morphological changes in these biopsy specimens and allograft function 1, 3 and 5 years after transplantation is assessed. A highly significant direct relationship exists between early graft failure and the presence of medial necrosis of arteries, acute glomerular lesions and interstitial haemorrhage. Less than 10% of grafts with one or more of these changes and none in which all three types of lesion were present were capable of supporting life at 1 year. There is a significant association between poor subsequent graft function and mononuclear cell infiltration of the intima of arteries. No clear relationship exists, however, between the function of grafts at 1 and 3 years and the degree of mononuclear cell infiltration of the interstitial tissue. Tubular necrosis was frequently observed and future graft performance is related to the extent and cause of the tubular damage.     

瓣下心肌可增强大鼠同种带瓣主动脉移植物免疫原性

对大鼠同种带瓣主动脉 (AVH)不同组织 (瓣下心肌、瓣叶、大动脉壁 )免疫原性进行比较 ,观察AVH移植后机体免疫排斥反应。采用大鼠AVH腹主动脉异位移植模型。实验分 3组 ,分别接受带瓣下心肌的AVH (A组 ,n =12 )、去除瓣下心肌的AVH(B组 ,n =12 )和单纯主动脉壁 (C组 ,n =12 )移植 ,以术前为对照。术后 2、4、8、12周测定血ICAM 1、TCR αβ、移植标本CD4 0 /CD4 0L的表达 ,并行组织病理学观察。 3组术后ICAM 1、TCR αβ和CD4 0 /CD4 0L的表达均比对照组明显增高 (P <0 0 5 )。A组术后 2～ 4周ICAM 1分别为 (9 0 32± 2 5 82 ) %和 (17 0 6±2 6 4 6 ) % ,明显高于B、C 2组 (P <0 0 1) ;术后 2周TCR αβ为 (4 1 5 9± 5 96 5 ) % ,亦明显高于B、C 2组 (P <0 0 5 ) ;术后 2周CD4 0表达为 4 333± 0 5 77,与C组相比差异明显 (P <0 0 5 )。病理学观察 3组均呈不同程度的免疫排斥反应表现 ,B、C 2组改变相似 ,A组病理改变比B、C 2组出现早、程度重。大鼠AVH移植诱发了机体免疫排斥反应 ,与瓣叶和动脉壁相比 ,瓣下心肌具有较强的免疫原性     

Morphometrics of corneal growth in chicks raised in constant light

In this study we wish to augment our understanding of the effect of environment on corneal growth and morphology. To understand how corneal development of chicks raised in constant light differs from that of ‘normal’ eyes exposed to cyclic periods of light and dark, white Leghorn chicks were raised under either constant light (approximately 700 lux at cage top) or in 12 h light/12 h dark conditions for up to 12 weeks after hatching. To determine whether corneal expansion is uniform, some birds from each group received corneal tattoos for periodic photographic assessment. By 16 days of age, constant light corneas weighed less than light/dark regimen corneas [7.39 ± 0.35 mg (SE) vs. 8.47 mg ± 0.26 mg SE wet weight, P ≤ 0.05], and corresponding differences were seen in corneal dry weights. Spatial expansion of the corneal surface was uniform in both groups, but the rate of expansion was slower in constant light chicks [0.0327 ± 0.009 (SE) vs. 0.144 ± 0.018 (SE) mm² day⁻¹ for normal chicks, P ≤ 0.001]. At 1 day of age, there were 422 ± 12.5 (SE) stromal cells 0.01 mm⁻² in the central cornea and 393 ± 21.5 (SE) stromal cells 0.01 mm⁻²peripherally. Although this difference is not statistically significant, the cell densities in the central cornea were always larger than those of the peripheral cornea in all eight measurements over a 10.5-week period, and this difference is significant (P ≤ 0.008, binomial test). Light/dark regimen birds show no such consistent difference in cell densities between central and peripheral corneas. Thus, the density distribution of corneal stromal cells of chicks grown in constant light differs from that of normal chicks. Taken together, all these observations suggest that diurnal cycles of light and darkness are necessary for normal corneal growth.     

Mutational spectrum of Korean patients with corneal dystrophy

Corneal dystrophy typically refers to a group of rare hereditary disorders with a heterogeneous genetic background. A comprehensive molecular genetic analysis was performed to characterize the genetic spectrum of corneal dystrophies in Korean patients. Patients with various corneal dystrophies underwent thorough ophthalmic examination, histopathologic examination, and Sanger sequencing. A total of 120 probands were included, with a mean age of 50 years (SD = 18 years) and 70% were female. A total of 26 mutations in five genes (14 clearly pathogenic and 12 likely pathogenic) were identified in 49 probands (41%). Epithelial–stromal TGFBI dystrophies, macular corneal dystrophy and Schnyder corneal dystrophy (SCD) showed 100% mutation detection rates, while endothelial corneal dystrophies showed lower detection rates of 3%. Twenty six non‐duplicate mutations including eight novel mutations were identified and mutations associated with SCD were identified genetically for the first time in this population. This study provides a comprehensive characterization of the genetic aberrations in Korean patients and also highlights the diagnostic value of molecular genetic analysis in corneal dystrophies.     

同种异体松质骨移植的生物力学研究

目的：探讨同种异体松质骨移植后的力学性能变化以及不同力学环境对其的影响。方法：在40只家兔前肢尺骨中段进行同种异体松质骨移植，并使左右侧植骨块分别承受正常生理载荷与低载荷，动物分批处死后取出植骨区标本进行骨密度值、三点弯曲、平均骨小梁厚度等测试。结果：同种异体松质骨愈合时的骨密度值、最大弯矩、平均骨小梁厚度逐渐上升。在移植后第16周时，和低载荷侧比较，正常载荷侧的上述指标都明显优于低载荷侧（P值分别＜0.01，0.05，0.05）。结论：同种异体松质骨移植后的力学性能变化与它所承受的载荷大小有较强的相关性。     

Genetics of the corneal endothelial dystrophies: an evidence‐based review

The aim of this review was to provide an evidenced‐based review of the genetic basis of the corneal endothelial dystrophies. A review of the English language peer‐reviewed literature describing the molecular genetic basis of posterior polymorphous corneal dystrophy (PPCD), congenital hereditary endothelial dystrophy (CHED), Fuchs endothelial corneal dystrophy (FECD) and X‐linked endothelial corneal dystrophy (XECD) was performed. Mutations in several genes have been implicated as playing a pathogenic role in the corneal endothelial dystrophies: VSX1 mutations in PPCD1; COL8A2 mutations in PPCD2 and FECD; ZEB1 mutations in PPCD3 and FECD; and SLC4A11 mutations in CHED2 and FECD. However, linkage, association and familial segregation analyses support a role of only one gene in each corneal endothelial dystrophy: ZEB1 in PPCD3, SLC4A11 in CHED2 and COL8A2 in FECD (early onset). In addition, insufficient evidence exists to consider the autosomal dominant form of CHED (CHED1) as distinct from PPCD. An accurate classification of the corneal endothelial dystrophies requires a critical review of the evidence to support the role of each suggested chromosomal locus, gene and genetic mutation associated with a corneal endothelial dystrophy. Only after the separation of evidence from opinion is performed can a critical examination of the molecular pathways that lead to endothelial dysfunction in each of these disorders be accurately performed.     

Allograft inflammatory factor-1 (AIF-1) is crucial for the survival and pro-inflammatory activity of macrophages

Our previous studies revealed that macrophages played an important role in linking injury, inflammatory and immune response in small-for-size liver transplantation. However, the molecular basis that promoted macrophage activation was not clear. In the present study, we explored the potential role of allograft inflammatory factor-1 (AIF-1) in mediating the survival and pro-inflammatory activity of macrophages in a macrophage cell line. First, the expression of AIF-1 was investigated with the stimulation of pro-inflammatory cytokines and anti-inflammatory treatment. Second, the level of inducible nitric oxide synthase (iNOS) and the survival and migration activity of macrophages were determined with the alterations of AIF-1 expression. Finally, a potential molecule that regulated AIF-1 expression was identified by the proteomic approach. The macrophage cell line expressed a certain level of endogenous AIF-1, which could be enhanced by pro-inflammatory cytokines IL-1beta or tumor necrosis factor-alpha and suppressed by anti-inflammatory drug sodium salicylate. AIF-1 augmentation induced by AIF-1/PCDNA3.1(+) transfection enhanced the levels of iNOS and monocyte chemoattractant protein-1, and promoted the cell migration. On the other hand, suppression of AIF-1 expression by AIF-1/short interference RNA (siRNA) inhibited iNOS production, induced macrophage cell apoptosis and blocked the cell migration. Using two-dimensional electrophoresis, a disintegrin and metalloproteinase domain 3 (ADAM3) was identified after AIF-1/siRNA transfection. Transfection of ADAM3/PCDNA3.1(+) up-regulated the expression of AIF-1 and iNOS, whereas suppression of ADAM3 expression down-regulated AIF-1 and iNOS expression. In conclusion, AIF-1 played an important role in the survival and pro-inflammatory activity of macrophages, and ADAM3 might be an upstream molecule that regulated AIF-1 expression.     

Rapidly proliferative arteriopathy in cyclosporin-induced permanently surviving rat cardiac allografts simulating chronic vascular rejection.

In an attempt to create a model of chronic rejection in rat, cardiac allograft transplantation was performed of hearts from PVG rats to DA rats treated with cyclosporin, 20 mg/kg body weight per day for 14 days and thereafter no immunosuppressive therapy. Experiments were made in 83 rats fed on a normal diet and in 24 rats fed on a diet containing an additional 0.5% cholesterol. Rats on the normal diet showed moderate signs of acute rejection during the first 20-40 days and grafts were lost in acute rejection during this period of time. However, after 2-3 months no signs of acute rejection were present. On the contrary, excessive proliferative changes of the vascular intima and endocardium along with fibrosis and fibrin deposition appeared and was progressive until 6 months post-transplantation. These morphological changes are similar to those found in chronically rejected organs like heart and kidney. In rats fed on a cholesterol diet after cessation of cyclosporin, development of the vascular and endocardial proliferative changes appeared three to four times as fast and were on average fully developed within 4-6 weeks post cessation of cyclosporin treatment. The recipients' own hearts showed no signs of vascular or endocardial damage. It is thus concluded that two models of vascular and endocardial proliferative changes in cardiac allografts have been developed showing distinct similarities to chronic vascular rejections seen in the clinical transplantation and with apparent similarities to severe arteriosclerosis. The models could be useful in the investigation of pathogenesis and therapeutical means for preventing chronic vascular damage in transplanted organs and arteriosclerosis.