Plant-derived angiogenin fusion protein's cytoprotective effect on trabecular meshwork damage induced by Benzalkonium chloride in mice.

BACKGROUND: Benzalkonium chloride (BAK), commonly used in glaucoma treatment, is an eye drop preservative with dose-dependent toxicity. Previous studies have observed the multi-functional benefits of angiogenin (ANG) against glaucoma. In our study, we evaluated ANG's cytoprotective effect on the trabecular meshwork (TM) damage induced by BAK. Additionally, we developed a plant-derived ANG fusion protein and evaluated its effect on TM structure and function. METHODS: We synthesized plant-derived ANG (ANG-FcK) by fuzing immunoglobulin G's Fc region and KDEL to conventional recombinant human ANG (Rh-ANG) purified from transgenic tobacco plants. We established a mouse model using BAK to look for degenerative changes in the TM, and to evaluate the protective effects of ANG-FcK and Rh-ANG. Intraocular pressure (IOP) was measured for 4 weeks and ultrastructural changes, deposition of fluorescent microbeads, type I and IV collagen, fibronectin, laminin and α-SMA expression were analyzed after the mice were euthanized. RESULTS: TM structural and functional degeneration were induced by 0.1% BAK instillation in mice. ANG co-treatment preserved TM outflow function, which we measured using IOP and a microbead tracer. ANG prevented phenotypic and ultrastructure changes, and that protective effect might be related to the anti-fibrosis mechanism. We observed a similar cytoprotective effect in the BAK-induced degenerative TM mouse model, suggesting that plant-derived ANG-FcK could be a promising glaucoma treatment.

Transplantation of autologous perichondrium with amniotic membrane for progressive scleral necrosis.

PURPOSE: Scleral necrosis with severe ischemia is refractory to conventional treatment because of avascular progressive necrosis. We assessed the therapeutic efficacy and safety of autologous perichondrium transplantation in patients with progressive scleral necrosis (PSN) and analyzed the clinical effects. METHODS: This study was a prospective, interventional, and noncomparative case series. Reconstructive surgery using autologous perichondrium and amniotic membrane (AM) was performed in patients with PSN who showed progressive ischemic scleral melting with impending perforation state and/or broad avascular area larger than 10 mm in diameter. The primary outcome was restoration of scleral integrity with healthy vascularized epithelium over the graft at six months after surgery. The secondary outcome was complication rate associated with autologous perichondrium graft use. RESULTS: Eighteen eyes of 14 patients underwent reconstructive surgery using autologous perichondrium patch and AM grafts. Observations indicated the graft provided the eyeball with successful structural integrity in 17 out of the 18 cases (94.4%) at six months after surgery. One eye showed a small scleral defect due to wound dehiscence at four month after the surgery. Additional surgery using perichondrium and AM stabilized the eye. The scleral necrosis healed completely after perichondrium and AM transplantation, even in cases with full-thickness scleral defect. The scleral integrity was maintained until the last follow-up session. There were no serious complications of endophthalmitis or graft infection. CONCLUSIONS: Reconstructive surgery using autologous perichondrium and AM is an effective method for restoration of scleral integrity and vascularization of the episclera and conjunctiva in eyes with PSN. Therefore, autologous perichondrium can be considered as an appropriate new biologic tissue for PSN.

Current approaches and future directions in the management of pterygium.

Pterygium, a common ocular surface disorder, has a complex pathophysiology that may mimic tumorigenesis. There is altered expression of cell cycle/proliferation-related factors in pterygium tissues. Therefore, similar to cancer treatments, the management of pterygium ought to be multifactorial based on the patient's condition. Current therapeutic methods for pterygium are focused on surgical resection in conjunction with antimetabolite use, in addition tissue graft is usually performed in the context of the avoidance of bare sclera. However, future directions in the management of pterygia will likely focus on genetic approaches. This perspective views the pathogenesis of pterygium, its existing therapies as well as current and future challenges in its treatment.

TNF-α upregulates HIF-1α expression in pterygium fibroblasts and enhances their susceptibility to VEGF independent of hypoxia.

The clinical manifestations of pterygium are characterized by rapid growth and postoperative recurrences. We had previously proposed that hypoxia-inducible factor (HIF)-1α recruits progenitor cells during the development and progression of pterygia. Recently, it was reported that various stimuli, including inflammation, could activate HIF-1α even under normoxic conditions. The ocular surface directly faces external environments, and is thus frequently exposed to inflammatory insults. First, we examined the gene expression of HIF-1α, its downstream molecule, vascular endothelial growth factor (VEGF)-A, and VEGF receptor (VEGFR)-2 in corneal and conjunctival cells compared with cultured human umbilical vein endothelial cells. Corneal fibroblasts had high expression of VEGFR-2 in the presence of TNF-α, and HIF-1α was activated by TNF-α in diverse ocular surface cells. The HIF-1α/VEGF/VEGFR signaling pathway in response to TNF-α was evaluated in cultured human pterygium fibroblasts (HPFs) at the gene and protein levels and was compared to treatment with cobalt chloride (CoCl2), a hypoxic mimetic, to exclude the effect of hypoxia. Although VEGF-A expression was not changed by TNF-α, expression of HIF-1α and VEGFR-2 was enhanced in HPFs treated with TNF-α, independent of hypoxia conditioning. In addition, VEGF-C gene expression was activated solely by TNF-α in HPF, but VEGF-B levels were not significantly affected. These results may provide mechanistic explanations for the uniquely vigorous proliferation of pterygium fibrovascular tissue during TNF-α-induced ocular surface inflammation.

Pulmonary glass particles may persist in the lung suppressing function of immune cells.

The health effects of silica may depend on the inherent properties of crystalline silica or on external factors affecting the biological activity or distribution of its polymorphs. Inhaled crystalline silica is classified as a Group I carcinogen, however, information on the health effects of amorphous silica is still insufficient. Considering that alveolar macrophages play a key role in both innate and adaptive immune responses for removal of foreign bodies that enter via the respiratory system, we treated sheet-like glass particles (SGPs), a type of noncrystalline amorphous silica, to MH-S cells, an alveolar macrophage cell line. SGPs reduced the generation of ROS and NO and induced cell death via multiple pathways. Although the expression of CD80, CD86, and CD40, increased by exposure to SGPs, the expression of MHC class II molecules had not notably changed. Additionally, expression of ICAM-1 tended to decrease. In mice, SGPs were distributed in the interstitial region of the lung without notable pathological lesion on day 14 after a single intratracheal instillation. Pulmonary total cell number increased significantly with the highest dose, but the levels of all measured inflammatory cytokines and chemokines, except IL-1, were lower in BAL fluid from SGP-treated mice compared to control. More interestingly, the expression of antigen presentation-related proteins was enhanced in the lungs of SGP-exposed mice concomitant with an increase in the number of mature dendritic cells, whereas the expression of ICAM-1, an important adhesion molecule for helper T cell recruitment, was suppressed. Taken together, we suggest that SGPs may induce adverse health effects by down-regulating function of immune cells in the lungs. Furthermore, ICAM-1 may play a key role in immune response to remove pulmonary SGPs.

Ribonuclease 5 facilitates corneal endothelial wound healing via activation of PI3-kinase/Akt pathway.

To maintain corneal transparency, corneal endothelial cells (CECs) exert a pump function against aqueous inflow. However, human CECs are arrested in the G1-phase and non-proliferative in vivo. Thus, treatment of corneal endothelial decompensation is limited to corneal transplantation, and grafts are vulnerable to immune rejection. Here, we show that ribonuclease (RNase) 5 is more highly expressed in normal human CECs compared to decompensated tissues. Furthermore, RNase 5 up-regulated survival of CECs and accelerated corneal endothelial wound healing in an in vitro wound of human CECs and an in vivo cryo-damaged rabbit model. RNase 5 treatment rapidly induced accumulation of cytoplasmic RNase 5 into the nucleus, and activated PI3-kinase/Akt pathway in human CECs. Moreover, inhibition of nuclear translocation of RNase 5 using neomycin reversed RNase 5-induced Akt activation. As a potential strategy for proliferation enhancement, RNase 5 increased the population of 5-bromo-2'-deoxyuridine (BrdU)-incorporated proliferating CECs with concomitant PI3-kinase/Akt activation, especially in CECs deprived of contact-inhibition. Specifically, RNase 5 suppressed p27 and up-regulated cyclin D1, D3, and E by activating PI3-kinase/Akt in CECs to initiate cell cycle progression. Together, our data indicate that RNase 5 facilitates corneal endothelial wound healing, and identify RNase 5 as a novel target for therapeutic exploitation.

Angiogenin ameliorates corneal opacity and neovascularization via regulating immune response in corneal fibroblasts.

BACKGROUND: Angiogenin (ANG), a component of tears, is involved in the innate immune system and is related with inflammatory disease. We investigated whether ANG has an immune modulatory function in human corneal fibroblasts (HCFs). METHODS: HCFs were cultured from excised corneal tissues. The gene or protein expression levels of interleukin (IL)-1beta (ß), IL-4, IL-6, IL-8, IL-10, complements, toll-like receptor (TLR)4, myeloid differentiation primary response gene (MYD)88, TANK-binding kinase (TBK)1, IkappaB kinase-epsilon (IKK-ε) and nuclear factor-kappaB (NF-κB) were analyzed with or without ANG treatment in tumor necrosis factor-alpha (TNF-α)- or lipopolysaccharide (LPS)-induced inflammatory HCFs by real-time polymerase chain reaction (PCR), Western blotting and immunocytochemistry. Inflammatory cytokine profiles with or without ANG were evaluated through immunodot blot analysis in inflammatory HCFs. Corneal neovascularization and opacity in a rat model of corneal alkali burn were evaluated after application of ANG eye drops. RESULTS: ANG decreased the mRNA levels of IL-1ß, IL-6, IL-8, TNF-α receptor (TNFR)1, 2, TLR4, MYD88, and complement components except for C1r and C1s and elevated the mRNA expression of IL-4 and IL-10. Increased signal intensity of IL-6, IL-8 and monocyte chemotactic protein (MCP)-1 and MCP-2 induced by TNF-α or LPS was weakened by ANG treatment. ANG reduced the protein levels of IKK-ε by either TNF-α and LPS, and decreased TBK1 production induced by TNF-α, but not induced by LPS. The expression of NF-κB in the nuclei was decreased after ANG treatment. ANG application lowered corneal neovascularization and opacity in rats compared to controls. CONCLUSION: These results demonstrate that ANG reduces the inflammatory response induced by TNF-α or LPS in HCFs through common suppression of IKK-ε-mediated activation of NF-κB. This may support the targeting of immune-mediated corneal inflammation by using ANG.

Angiogenin for the Diagnosis and Grading of Dry Eye Syndrome.

PURPOSE: To investigate the properties of angiogenin (ANG) as a potential tool for the diagnosis and grading of dry eye syndrome (DES) by analyzing tear protein profiles. METHODS: Tear samples were collected with capillary tubes from 52 DES patients and 29 normal individuals as controls. Tear protein profiles were analyzed with an immunodot blot assay as a screening test. To confirm that the tear ANG levels were in inverse proportion to the disease severity grade, the ANG and lactoferrin (LF) tear contents of normal controls and DES patients were compared in an enzyme-linked immunosorbent assay. RESULTS: In the immunodot blot assay, the ANG area was lower in patients with grades 3 and 4 DES than in normal controls. The areas of basic fibroblast growth factor, transforming growth factor ß2, and interleukin 10 were significantly greater than those of normal controls only in grade 4 DES patients, but these proteins were not linearly correlated with dry eye severity. Upon enzyme-linked immunosorbent assay analysis, the mean concentrations of ANG and LF decreased significantly as dry eye severity increased, except between grades 1 and 2. In addition, the ratios of ANG and LF to total tear proteins were correlated significantly with DES severity. CONCLUSIONS: ANG level was significantly lower in DES patients than in normal controls, and was significantly correlated with the worsening severity of DES, except between grades 1 and 2, as was LF. Therefore, ANG may be a useful measure of DES severity through proteomic analysis.

Oncologic results and prognostic predictors of patients with locally advanced rectal cancer showing ypN0 after radical surgery following neoadjuvant chemoradiotherapy.

PURPOSE: Adjuvant chemotherapy is recommended for patients with locally advanced rectal cancer after radical surgery following neoadjuvant chemoradiotherapy (NCRT) regardless of the final pathologic stage. However, the efficacy of adjuvant chemotherapy in ypN0 patients remains controversial. The aim of this study was to evaluate the oncologic outcomes and analyze the prognostic factors for ypN0 patients in order to estimate prognosis and establish an effective adjuvant chemotherapy strategy for stage 0-II rectal cancers after radical surgery following NCRT. METHODS: Between January 1999 and December 2009, the medical records of 202 patients who had been diagnosed with locally advanced rectal cancer, underwent radical surgery following NCRT, and showed ypN0 were retrospectively reviewed. RESULTS: The median follow-up period was 60.5 months. The 5-year local recurrence rate was 3.1 %. The 5-year disease-free survival and 5-year overall survival were 86.3 and 86.9 %. Postirradiation T3-4 and abdominoperineal resection (APR) were the independent prognostic indicators for disease-free survival (p = 0.001, p = 0.003) and overall survival (p = 0.001, p = 0.002). Adjuvant chemotherapy improved local recurrence in the patient with ypT3-4 and patients who had undergone APR (p = 0.014, p = 0.002). APR affected local recurrence, disease-free survival, and overall survival of ypT3-4 patients (p = 0.013. 0.029, and 0.001) CONCLUSIONS: Postirradiation T3-4 and APR are the significant prognostic factors for ypN0. Further randomized prospective study is needed to evaluate the oncologic benefit of adjuvant chemotherapy in ypN0 patients, especially those with ypT3-4 and those having undergone APR, and to confirm which chemotherapeutic agent could improve the oncologic outcomes of patients poorly responding to NCRT.

Deep anterior lamellar keratoplasty using irradiated acellular cornea with amniotic membrane transplantation for intractable ocular surface diseases.

PURPOSE: To report the clinical outcomes of deep anterior lamellar keratoplasty (DALK) when sterile gamma-irradiated acellular corneal tissues (VisionGraft) are used in combination with amniotic membrane transplantation (AMT) for intractable ocular surface diseases. METHODS: The medical records of fifteen patients who had DALK with AMT were retrospectively reviewed. Indications for surgery included ocular burn, bacterial keratitis, herpes simplex virus keratitis, corneal opacity with Stevens-Johnson syndrome, Mooren's ulcer, idiopathic myxoid degeneration of corneal stroma, and recurrent band keratopathy. DALK was performed using partial-thickness acellular corneal tissue and a temporary amniotic membrane patch was added at the end of the operation. RESULTS: All cases that underwent DALK with AMT became epithelialized within 2 postoperative weeks. Twelve patients showed favorable outcomes without graft rejection, corneal opacification, or neovascularization. The other three grafts developed corneal opacification and neovascularization, and required additional penetrating keratoplasty (PK). Unlike the results of previous PKs, there were no graft rejections and the graft clarity was well-maintained in these three cases for at least 8 months after PK. CONCLUSIONS: DALK using sterile acellular corneal tissues in combination with AMT may be a good therapeutic strategy for treating intractable ocular surface diseases because of lowered immune rejection, fibroblast activation, and facilitation of epithelialization. Furthermore, DALK can help stabilize the ocular surface, prolong graft survival, and may allow better outcomes when combined with subsequent PK.

Influence of congenital facial nerve palsy on craniofacial growth in craniofacial microsomia.

Facial muscles are of major importance in human craniofacial growth and development. The purpose of our study was to investigate whether congenital facial nerve palsy influences craniofacial growth in craniofacial microsomia. Fifty-one patients with unilateral craniofacial microsomia and no history of craniofacial skeletal surgery whose radiographs were taken after craniofacial growth was complete were included in this study. These patients were divided into groups in which the facial nerve was involved or uninvolved. The authors evaluated a total of seven measurement items to analyze the midface and mandibular asymmetry. Twenty patients had facial nerve involvement, and 31 had no involvement. None of the measurement items revealed any significant differences between the facial nerve-involved group and the uninvolved group within the same modified Pruzansky grade. There was no correlation between the type of facial nerve involvement and the measurement items. In relationships among the measurement items within each group, maxillary asymmetry was indirectly correlated with mandibular asymmetry or midline deviation through the occlusal plane angle in the uninvolved groups. However, in the facial nerve-involved group, the relationships disappeared. When the correlations in the facial nerve-involved group were compared with those of the uninvolved group, the relationships in the uninvolved group appeared more significant than in the facial nerve-involved group. The loss of relationships between the upper and lower jaw in the facial nerve-involved group might have been caused by subtle changes, which occur in midfacial bones and in the mandible due to facial nerve palsy. The main limitation of our study is that aside from facial nerve palsy, craniofacial microsomia has many factors that can influence craniofacial growth, such as hypoplasia of the mandibular condyle and soft tissue deficiencies.

Evaluation of stem cell components in retrocorneal membranes.

The purpose of this study was to elucidate the origin and cellular composition of retrocorneal membranes (RCMs) associated with chemical burns using immunohistochemical staining for primitive cell markers. Six cases of RCMs were collected during penetrating keratoplasty. We examined RCMs with hematoxylin and eosin (H&E), periodic acid-Schiff (PAS) staining and immunohistochemical analysis using monoclonal antibodies against hematopoietic stem cells (CD34, CD133, c-kit), mesenchymal stem cells (beta-1-integrin, TGF-ß, vimentin, hSTRO-1), fibroblasts (FGF-ß, α-smooth muscle actin), and corneal endothelial cells (type IV collagen, CD133, VEGF, VEGFR1). Histologic analysis of RCMs revealed an organized assembly of spindle-shaped cells, pigment-laden cells, and thin collagenous matrix structures. RCMs were positive for markers of mesenchymal stem cells including beta-1-integrin, TGF-ß, vimentin, and hSTRO-1. Fibroblast markers were also positive, including FGF-ß and α-smooth muscle actin (SMA). In contrast, immunohistochemical staining was negative for hematopoietic stem cell markers including CD34, CD133 and c-kit as well as corneal endothelial cell markers such as type IV collagen, CD133 except VEGF and VEGFR1. Pigment-laden cells did not stain with any antibodies. The results of this study suggest that RCMs consist of a thin collagen matrix and fibroblast-like cells and may be a possible neogenetic structure produced from a lineage of bone marrow-derived mesenchymal stem cells.

Angiogenin reduces immune inflammation via inhibition of TANK-binding kinase 1 expression in human corneal fibroblast cells.

Angiogenin (ANG) is reportedly multifunctional, with roles in angiogenesis and autoimmune diseases. This protein is involved in the innate immune system and has been implicated in several inflammatory diseases. Although ANG may be involved in the anti-inflammatory response, there is no evidence that it has direct anti-inflammatory effects. In this study we sought to determine whether ANG has an anti-inflammatory effect in human corneal fibroblasts (HCFs) exposed to media containing tumor necrosis factor-alpha (TNF-α). We found that ANG reduced the mRNA expression of interleukin-1 beta (IL-1ß), -6, -8 and TNF-α receptors (TNFR) 1 and 2. In contrast, ANG increased the mRNA expression of IL-4 and -10. Protein levels of TANK-binding kinase 1 (TBK1) were reduced by ANG in HCFs treated with TNF-α. Moreover, ANG diminished the expression of IL-6 and -8 and monocyte chemotactic protein- (MCP-) 1. The protein expression of nuclear factor-κB (NF-κB) was downregulated by ANG treatment. These findings suggest that ANG suppressed the TNF-α-induced inflammatory response in HCFs through inhibition of TBK1-mediated NF-κB nuclear translocation. These novel results are likely to play a significant role in the selection of immune-mediated inflammatory therapeutic targets and may shed light on the pathogenesis of immune-mediated inflammatory diseases.

Treatment of an 8-mm myxoma using acellular corneal tissue.

A myxoma is a benign tumor found in the heart and in various soft tissues; however, a corneal myxoma is rare. A mucinous mass of unknown etiology was observed on the left cornea of a 32-year-old male patient. We performed deep anterior lamellar keratoplasty using acellular corneal tissue and concurrent amniotic membrane transplantation. Hematoxylin and eosin staining revealed vacuolation of the parenchyma and myxoid change in the corneal tissue that occurred in the anterior half of the corneal parenchyma. We identified a myxoid stroma by Alcian blue staining and observed collagen fibers with denatured stroma by Masson trichrome staining. The patient's visual acuity improved from light perception to 20 / 200, and the intraocular pressure remained within the normal range for one year after surgery. The transplanted cornea survived successfully with well-maintained transparency, and recurrence was not observed one year after surgery.

Influence of modified Furlow double opposing Z-plasty on mandibular growth in Oriental patients with cleft palate and/or lip.

The purpose of this study was to statistically analyze mandibular growth disturbance in the Oriental cleft population and compare this with that of non-cleft children through cephalometric analysis. Thirty-six children with cleft palates, repaired using a modification of Furlow double opposing Z-plasty, were chosen. Comparative analyses among 3 types of cleft were performed. Statistical analyses of 8 linear and angular measurements were performed in cleft patients and the non-cleft population using Fisher Z-transformation. Comparative analysis showed no significant difference among the 3 types of cleft. In the Oriental cleft group treated with modified double opposing Z-plasty, the spatial position of the mandible showed significant differences compared with the non-cleft group. A backward inclination of the anterior surface of the mandible and downward rotation of the mandibular body were identified. Some of our results regarding gonial angle and length of the mandibular body conflicted with previous Western studies.

Zinc finger protein in severe dry eye syndrome.

PURPOSE: Zinc finger protein known to induce squamous metaplasia and regulate vitamin A expression has been few investigated as tear protein. We investigated tear protein variations in patients with dry eye syndrome (DES). MATERIALS AND METHODS: Tears from healthy subjects as control and patients with DES were collected. Tear proteins were separated by one-dimensional electrophoresis. The protein bands were analyzed by nano liquid chromatography-mass spectrometry. RESULTS: Compared to the controls, significant down-regulation of lactoferrin and lysozyme was detected, while significant up-regulation was observed for serum albumin in patients with DES. DES grade 4 patients showed different protein patterns. Zinc-finger motif-enhancer binding-protein-1 gamma and bromodomain adjacent to zinc finger domain 2B were detected in DES grade 4 patients. CONCLUSIONS: Tear protein changes are valuable to diagnosis DES. Zinc finger proteins may be associated with pathophysiology of severe DES. Further studies are needed to better understand the relationship of zinc finger proteins in tear of patients with DES.

Autologous tragal perichondrium patch graft for ahmed glaucoma valve tube exposure.

PURPOSE: To describe a novel method of repair for an exposed Ahmed valve tube using tragal perichondrium. METHODS: A 56-year-old man with a severe chemical burn was referred to our clinic for the management of total limbal stem cell deficiency. During the treatment, an Ahmed valve tube was implanted to control refractory glaucoma. Tube exposure occurred 39 months after implantation. A glycerin-preserved donor sclera patch graft was used to cover the exposed lesion. However, 21 months later, the tube became exposed again. Autologous tragal perichondrium was then harvested and used to repair the exposed tube. RESULTS: The exposed tube was closed successfully without recurrence, graft thinning, or graft-related infection or inflammation over a 20-month follow-up period. CONCLUSIONS: Perichondrial patch grafts can be a useful alternative for repair of exposed glaucoma drainage tubes.

Mandibular distraction in unilateral craniofacial microsomia: longitudinal results until the completion of growth.

BACKGROUND: The purpose of this study was to evaluate the results of long-term follow-up in patients with relatively severe unilateral craniofacial microsomia after mandibular distraction. METHODS: The sample consisted of 26 patients with an average age of 6.08 years at the time of distraction. All patients had nonsyndromic unilateral craniofacial microsomia (Pruzansky-Kaban types IIA, IIB, and III). Follow-up for all patients continued until the completion of growth. The 26 clinical records and posteroanterior cephalograms of the patients, taken before distraction (time 0) and approximately 1, 4, and 11 years (time 1, time 2, and time 3, respectively) after distraction, were used. Four items (i.e., supraorbital tilting angle and occlusal tilting angle to the horizontal reference line, and maxillary and mandibular tilting angles to the vertical reference line) were analyzed at each of the four time intervals. RESULTS: The overall mean distraction amount was 23 mm vertically and 21 mm horizontally. The average cephalometric follow-up was 131.7 months. In this study, the mandibular horizontal and vertical changes after distraction showed a gradual return of the asymmetry with growth in all patients. There were no statistically significant differences in any of the analyzed angles between the preoperative and final records, except for the supraorbital tilting angle. CONCLUSIONS: The longitudinal results achieved by distraction osteogenesis were unstable and generally led to relapse, although they were very good in the early postoperative period. Therefore, further efforts are suggested to find a solution that prevents relapse. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, IV.

Overexpression of angiogenin in pterygium body fibroblasts and its association with proliferative potency.

PURPOSE: Angiogenin (ANG) originally was identified as an angiogenic tumor factor, and recently its biologic activity is extended to stimulating cell proliferation. With viewing pterygium as a tumorigenic mimicry, we investigated ANG profiles within pterygia. METHODS: Expression levels of ANG were assessed using immunohistochemistry, RT-PCR, and Western blotting through examination of excised specimens and cultured fibroblasts from pterygium and conjunctiva tissues. The phenotypes of pterygia were classified by four grading indices, including recurrence, growth activity, pterygium body translucency (T), and vascularity (V). Then, ANG levels in pterygia were differentiated according to phenotypes of pterygia, and were compared to levels in normal conjunctiva. Furthermore, to investigate ANG-related acquisition of proliferative potency in fibroblasts, the correlation between ANG and α-smooth muscle actin (α-SMA) levels was evaluated. RESULTS: In immunohistochemistry, ANG was expressed strongly in pterygium stroma with all four severe phenotypes (with recurrence, active growth, thick body [T3], and marked vascularization [V3]), especially at the perivascular areas. There was a trend toward higher ANG expression in cultured fibroblasts of pterygia with severe phenotype, compared to those without and with normal conjunctiva. However, pterygium body V had a weak association with ANG expression. Additionally, Western blotting revealed a significant positive correlation between the expression levels of α-SMA and ANG. CONCLUSIONS: Overexpression of ANG in pterygium body fibroblasts might be involved in active pterygium growth with thick pterygium body formation and increased risk of recurrence. A possible mechanism for this finding includes ANG-related transition of pterygium fibroblasts to the proliferative state.

The results and complications of cranial bone reconstruction in patients with craniosynostosis.

The aim of this study was to report the treatment results and complications of cranial bone reconstruction in patients with craniosynostosis at the Seoul National University Children's Hospital over the 14-year period from 1996 to 2009. A retrospective study was undertaken on 96 cases of 94 consecutive patients diagnosed with craniosynostosis, operated on between 1996 and 2009. The authors collected the data regarding age at surgery, operative time, duration of hospitalization, and follow-up period. Surgical results including the amount of blood loss, signs related to increased intracranial pressure, aesthetic results, cranial index, and complications were evaluated. The authors compared these results with those of the past 10-year period from 1986 to 1995 at the same hospital. There were 81 patients with single synostosis and 13 patients with multiple synostoses. The age of patients ranged from 3 months to 130 months (mean, 27.9 months). The mean operative time was 7.54 ± 1.77 hours. The duration of hospital stay ranged from 5 to 70 days with a mean period of 8.97 days. The intraoperative loss of hemoglobin ranged from 0 to 9.2 g/dL (mean, 2.35 g/dL). The symptoms of increased intracranial pressure were improved after cranioplasty. With regard to aesthetic results, 94 patients were classified into category I and 2 into category IV. The cranial index decreased from 88.0% ± 1.2% to 86.9% ± 1.1% on average. There were 34 minor complications in 28 operations (29.2%). There were statistically significant differences in age at surgery, operative time, duration of hospital stay, and reoperation rate between the recent 14-year study and the past 10-year study. We observed good surgical results and low complications of cranial bone reconstruction in patients with craniosynostosis. The results of the recent 14-year operations were better than those of the past 10-year operations.