Reconstruction of the ocular surface by autologous transplantation of rabbit adipose tissue-derived stem cells on amniotic membrane.

BACKGROUND: Corneal disease is the second most common cause of blindness in China. Clinically, treatment options for corneal diseases with limbal stem cell deficiency (LSCD) are limited due to a shortage of organ donors and inevitable immune rejection. This study aims to determine the efficacy of reconstructing the ocular surface using autologous cultivated adipose tissue-derived stem cells (ADSCs) and to develop a new clinical treatment for patients with LSCD. METHODS: A rabbit LSCD model was first established. Two weeks later, the animals were divided into three groups, including the sham group, the amniotic membrane transplantation group, and the ADSC combined with amniotic membrane transplantation group, and underwent surgery. The efficacy of reconstructing the ocular surface using ADSCs was evaluated using immunofluorescent staining, confocal microscopy (CM) observation, H&E staining, immunohistochemical staining, and scanning transmission electron microscopy observation one, two and four weeks after surgery. RESULTS: Evaluations of immunofluorescent staining of the cornea pre- and post-surgery yielded significantly lower scores for the corneas in the ADSCs transplantation group than for those in the sham group (F=-7, P=0.002, <0.05) and the amniotic membrane transplantation group (F=-4.67, P=0.018, <0.05) two weeks after surgery. Four weeks after surgery, the corneas of the ADSC combined with amniotic membrane transplantation group were scored significantly lower than those in the sham group (F=-8, P=0.007, <0.05) and the amniotic membrane transplantation group (F=-5.33, P=0.046, <0.05). The data suggest that the use of ADSCs to treat LSCD showed greater efficacy than the other treatment methods. The growth of ADSCs on the corneal surface was examined using confocal and electron microscopes. K3/K12 expression in the corneal epithelium, which was reconstructed by ADSCs, was negative, as shown by immunohistochemical staining. CONCLUSIONS: Ocular surface reconstruction can be improved by using ADSCs as seed cells and the amniotic membrane as a carrier, thus providing a new therapeutic strategy for patients with LSCD.

Metabolomic analysis of the changes of pathways in immunosuppressive mechanism caused by cyclophosphamide in mice / 中国实验动物学报

Objective To investigate the changes of metabolic pathways in the process of immunesuppression in mice caused by cyclophosphamide by metabolomic analysis. Methods Flow cytometry was used to detect the changes of immune cells, and the results were analyzed using partial least squares model. The potential biomarkers were screened by variable importance in projection ( VIP) parameters in the partial least squares model, and the differential metabolites were determined by statistical analysis. The differential metabolites were identified using the metabolomics rapid identification and analysis software. Nine kinds of metabolites were identified by searching and calculation, and result of pathway enrich-ment showed three differential metabolic pathways. Results Cyclophosphamide had a significant effect on the biosynthesis of unsaturated fatty acids, metabolism of mitochondrial fatty acids, metabolism of glycophospholipid, biosynthesis of steroid hormones, metabolism of arachidonic acid, metabolism of fatty acids, and the biosynthesis of pyrimidine. Conclusions The most important metabolic pathways affected by cyclophosphamide in the normal body are the metabolism of arachidonic acid, glycophospholipid and pyrimidine.

Establishment and evaluation of a guinea pig model of itching / 中国实验动物学报

Objective To optimize the optimal doses of histamine and 4-aminopyridine (4-AP) in the establish-ment of guinea pigs models of itching, and to establish a new guinea pig model of itching. Methods The central composite design-response surface method was used to arrange the experiment. In the experiment different pruritus agents were hypo-dermically injection of 0. 5 mL in the depilated area, and the scratching incubation period and scratching number in 30 mi-nutes were counted after the injection. The guinea pig itching model was evaluated by observing the behavioral changes of guinea pigs and measuring the levels of histamine and interleukin-6 in the blood. Results The behavioral experiments found that the scratching frequency in the the combination group was significantly higher than the histamine group and 4-AP group (P<0. 01). The itching latency of the combination group was significantly shorter than that of the histamine group and 4-AP group (P<0. 01). Compared with the control group, the histamine concentrations of the combination group and histamine group were significantly increased ( P<0. 05 or P<0. 01 ) , and the level of the combination group was lower than that of the histamine group (P<0. 05). Compared with the control group, the serum IL-6 concentrations of histamine group, 4-AP group and combination group were significantly higher (P<0. 01 or P<0. 05), and those in the combination group were significantly higher than the histamine and 4-AP groups. Compared with the control group, pathologic examina-tion showed proliferation of inflammatory cells in all model groups, and the reaction of the combination group was more ob-vious. Conclusions The optimal conditions used in this experiment are easy to achieve and have good reproducibility in the establishment of a guinea pig model of itching.

Metabolomic analysis of the changes of pathways in immunosuppressive mechanism caused by cyclophosphamide in mice / 中国实验动物学报

Objective To investigate the changes of metabolic pathways in the process of immunesuppression in mice caused by cyclophosphamide by metabolomic analysis. Methods Flow cytometry was used to detect the changes of immune cells, and the results were analyzed using partial least squares model. The potential biomarkers were screened by variable importance in projection ( VIP) parameters in the partial least squares model, and the differential metabolites were determined by statistical analysis. The differential metabolites were identified using the metabolomics rapid identification and analysis software. Nine kinds of metabolites were identified by searching and calculation, and result of pathway enrich-ment showed three differential metabolic pathways. Results Cyclophosphamide had a significant effect on the biosynthesis of unsaturated fatty acids, metabolism of mitochondrial fatty acids, metabolism of glycophospholipid, biosynthesis of steroid hormones, metabolism of arachidonic acid, metabolism of fatty acids, and the biosynthesis of pyrimidine. Conclusions The most important metabolic pathways affected by cyclophosphamide in the normal body are the metabolism of arachidonic acid, glycophospholipid and pyrimidine.

Establishment and evaluation of a guinea pig model of itching / 中国实验动物学报

Objective To optimize the optimal doses of histamine and 4-aminopyridine (4-AP) in the establish-ment of guinea pigs models of itching, and to establish a new guinea pig model of itching. Methods The central composite design-response surface method was used to arrange the experiment. In the experiment different pruritus agents were hypo-dermically injection of 0. 5 mL in the depilated area, and the scratching incubation period and scratching number in 30 mi-nutes were counted after the injection. The guinea pig itching model was evaluated by observing the behavioral changes of guinea pigs and measuring the levels of histamine and interleukin-6 in the blood. Results The behavioral experiments found that the scratching frequency in the the combination group was significantly higher than the histamine group and 4-AP group (P<0. 01). The itching latency of the combination group was significantly shorter than that of the histamine group and 4-AP group (P<0. 01). Compared with the control group, the histamine concentrations of the combination group and histamine group were significantly increased ( P<0. 05 or P<0. 01 ) , and the level of the combination group was lower than that of the histamine group (P<0. 05). Compared with the control group, the serum IL-6 concentrations of histamine group, 4-AP group and combination group were significantly higher (P<0. 01 or P<0. 05), and those in the combination group were significantly higher than the histamine and 4-AP groups. Compared with the control group, pathologic examina-tion showed proliferation of inflammatory cells in all model groups, and the reaction of the combination group was more ob-vious. Conclusions The optimal conditions used in this experiment are easy to achieve and have good reproducibility in the establishment of a guinea pig model of itching.

Is glaucoma comprehension associated with psychological disturbance and vision-related quality of life for patients with glaucoma? A cross-sectional study.

OBJECTIVE: To investigate whether understanding of glaucoma is associated with psychological disturbance and vision-related quality of life (QoL) in patients with glaucoma. DESIGN: A prospective, hospital-based, cross-sectional study. SETTING: Shanghai, China. PARTICIPANTS: Five hundred patients with glaucoma were included in the study. Participants were asked to complete the glaucoma comprehension questionnaire, the Hospital Anxiety and Depression Scale (HADS) questionnaire, and the 25-item National Eye Institute Visual Functioning Questionnaire (NEI VFQ-25). OUTCOME MEASURES: Sociodemographic data and clinical data were collected. Multivariate adjusted linear regression analyses were performed to examine factors related to psychological disorder and QoL. The relationship between understanding of glaucoma and psychological status or QoL was analysed. RESULTS: The mean age of patients was 56.5±17.2 years, and the mean duration of glaucoma was 4.6±6.3 years. About half (54.2%) of the patients had primary open angle glaucoma, 37.2% had primary angle closure glaucoma, and 8.6% had secondary glaucoma. The total score and the subscores of knowledge of glaucoma, compliance and habits were all negatively correlated with the HADS-Depression and HADS scores. For NEI VFQ-25, the total comprehension score was positively related to general vision, near activity, distance activity, social function, colour vision and the composite score. Habits had a significant influence on both HADS-Depression and the composite score of NEI VFQ-25. CONCLUSIONS: The level of understanding about glaucoma is an independent factor negatively associated with psychological disturbance and positively associated with QoL in patients with glaucoma. However, it is a subjective factor which can be regulated, so imparting knowledge about glaucoma and establishing appropriate habits might help patients alleviate psychological disturbance and enhance their QoL.

[Comparison of manual versus automatic titration in pressure determination for long-term therapy of continuous positive airway pressure in patients with obstructive sleep apnea hypopnea syndrome].

OBJECTIVE: To compare the continuous positive airway pressure (CPAP) of automatic titration with that of manual titration. METHODS: A total of 58 patients with obstructive sleep apnea and hypopnea syndrome (OSAHS) diagnosed by overnight polysomnography at sleep center of First Affiliated Hospital, Guangzhou Medical University were studied between December 2010 and December 2012. Manual titration was performed under full polysmnography and auto-titration at home for 3-7 nights. RESULTS: There were 52 males and 6 females with an age range of (48 ± 11) years. CPAP pressure titrated by automatic device (10.0 ± 2.2) cm H2O (1 cm H2O = 0.098 kPa) was significantly higher than that titrated manually (7.5 ± 1.5) cm H2O (P = 0.000). Apnea-hyponea index decreased significantly from (54.0 ± 21.0) events/h pre-treatment to (3.8 ± 2.5) events/h post-treatment under manual titration (P < 0.01). CONCLUSIONS: CPAP pressure titrated by automatic device is usually higher than that titrated manually. Manual titration should be performed if a patient can not tolerate the CPAP pressure titrated by an automatic device.

[Morphological characteristics of cornea in patients with vernal keratoconjunctivitis by in vivo laser scanning confocal microscopy].

OBJECTIVE: To explore the morphological characteristics on cornea in patients with vernal keratoconjunctivitis (VKC) by the application of in vivo laser scanning confocal microscopy (LSCM). METHODS: The experimental design was retrospective observation case series (case control study). Twenty-six patients, each diagnosed as bilateral VKC, were enrolled in the study, among which 13 were tarsal form, 5 were bulbar form and the rest were mixed form. Nine patients had the clinical course less than one year, eight subjects longer than three years, and the rest between them. Another twenty-six healthy volunteers with matching age and gender were selected as normal control. All participants had their right eyes examined with the in vivo confocal microscopy (HRT II/RCM). Central cornea and superior peripheral cornea were chosen as the examination points. The images were recorded automatically and cellular density of each layer was analyzed by installed software. Software ImageJ was utilized to analyze the density, diameter, branch number and tortuosity of subbasal nerve fiber in VKC patients. Independent t test was performed to assess the differences on cellular density between VKC patients and normal control, as well as those between central and peripheral cornea in VKC patients. Fisher chi-square test was used to compare the infiltration rate of Langerhans cells in corneal epithelium between VKC patients and controls. ANOVA was applied to assess the differences in cellular density among three subtypes, as well as among different duration of VKC. Independent t-test and chi-square test were applied to analyze the parameters of subbasal nerve fiber. RESULTS: The morphological changes in cornea included the absence of superficial hyperreflective polygonal epithelial cells, infiltration of Langerhans cells in and(or) underneath corneal epithelium and activation of keratocytes in anterior stroma. Corneal epithelium conjunctivalization and stromal neovascularization could be identified in patients with corneal neovascular epithelium. Longitudinal or oblique dark striae could be found in the posterior stroma in patients with complicated keratoconus. The density of epithelial cells at central and peripheral cornea in healthy controls were (6033.1 ± 998.7) cells/mm(2) and (6098.4 ± 298.3) cells/mm(2), while that in VKC patients were (5972.2 ± 1148.2) cells/mm(2) and (6178.5 ± 318.9) cells/mm(2) respectively, the differences being no statistical significant between them (t = 1.191, 1.011; P = 0.238, 0.318). However, it's found in VKC patients that cellular density at peripheral cornea was significantly higher than that at central area (t = 2.249, P = 0.03). The density of anterior stromal cells at central and peripheral cornea in healthy controls was (1001.4 ± 125.3) cells/mm(2) and (924.6 ± 201.4) cells/mm(2), while that in VKC patients was (1184.5 ± 115.3) cells/mm(2) and (1101.4 ± 151.1) cells/mm(2), the difference bearing no statistical significance (t = 6.617, 3.439; P = 0.001). The density of posterior stromal cells in normal subjects and VKC patients was (537.7 ± 42.6) cells/mm(2) and (548.7 ± 79.8) cells/mm(2), that of endothelial cells was (2985.7 ± 401.2) cells/mm(2) and (3021.5 ± 383.3) cells/mm(2), respectively, neither difference had statistical significance (t = 0.174, 1.112; P = 0.864, 0.282). Langerhans cell infiltration could be identified in 61.5% (16 cases) VKC patients, which was significantly higher than normal control (2 cases, 7.7%) (χ(2) = 12.49, P = 0.001). Furthermore, much intense Langerhans cells infiltration was found in bulbar form and mix form than tarsal form. (t = 6.617, P = 0.001). The density and diameter of subbasal nerve fiber in VKC patients decreased significantly than those in normal subjects, whereas the tortuosity increased significantly. CONCLUSIONS: The morphological changes of cornea in VKC patients mainly involve corneal epithelium, subbasal nerve fiber and anterior stroma. In vivo LSCM is helpful in discriminating the subtypes of VKC.

[Clinical applications and limitations of anterior segment optical coherence tomography].

Optical coherence tomography (OCT) is being employed more and more often to facilitate the diagnosis and management of the anterior eye segment diseases. The cross-sectional imaging capability of its higher resolution allows OCT to measure and visualize clear anatomic structures, specifically in anterior chamber biometry, corneal pachymetric mapping, and anterior chamber angle. Fourier-domain OCT technique which achieves higher speed and higher resolution and time-domain OCT technique have driven their further application in ophthalmological practice. In this review, we describe the principles and characteristics of OCT, summarize the recent utility and the limitations of anterior segment OCT in the pathologies and surgical planning of anterior chamber, cornea and surrounding areas, and prospect its future development.

[The ocular surface of severe alkali burns patients on confocal microscopy].

OBJECTIVE: To analyze the morphology on the ocular surface of severe alkali burns patients by in vivo laser scanning confocal microscopy. METHODS: This research was a retrospective observation case series. From February to November 2008 in Eye Ear Nose and Throat Hospital of Fudan University, 39 alkali burns patients who classified as III or IV according to Roper-Hall classification were enrolled in this study. They were divided into four groups according to the course of disease: A (less than 3 months), B (3 - 6 months), C (6 - 12 months) and D (over 12 months). In vivo laser scanning confocal microscopic examinations were performed on the injured cornea, the limbus and the bulbar conjunctiva and the images were recorded. The morphology of the injured cornea, the limbus and the bulbar conjunctiva was analyzed and the densities of the inflammatory cells and dendritic cells in the limbus were calculated. One-way analysis of variance was used to compare the means of the inflammatory cells and dendritic cells. Subsequently the data between two groups were analyzed by least significant difference. RESULTS: The corneal epitheliums of the patients in Group A manifested large irregular features with hyperreflective cytoplasm and hyporeflective nuclei, sometimes losing cell features. There were numerous small hyperreflective inflammatory cells in groups beneath the superficial epitheliums. Shallow corneal stroma was edema, and it was hard to discriminate the morphology of the stromal cells. Deep stromal cells were in the activated state. The images of the endothelial layer were unclear. In Group B and Group C, there were the same manifestation of the superficial epitheliums as the group A and it disappeared in Group D. The inflammatory cells beneath the superficial epitheliums reduced and some residual basal epitheliums and hyperreflective conjunctiva-like epitheliums were visible in Group B and Group C. In Group D, there were small oval tight-arranged cells with punctiform hyperreflective nuclei instead of normal corneal basal epitheliums. In Group B, it was still hard to discriminate the morphology of the shallow stroma cells. Deep stromal cells were still in the activated state. In Group C and Group D, corneal stroma was replaced by the fibrous tissues. The images of the endothelial layer were still unclear in the other groups. The Vogt palisades in the limbus of the severe alkali burns patients were destroyed in all groups. There were rich vascular nets in the limbus. The densities of the limbal inflammatory cells in four groups were (4023 +/- 343), (2975 +/- 246), (2652 +/- 375), (2679 +/- 299) cells/mm(2), respectively. Significant difference in inflammatory cell density was found among groups (F = 40.001, P = 0.000). The densities of the limbal dendritic cells in four groups were (106 +/- 19), (132 +/- 35), (141 +/- 26), (98 +/- 24) cells/mm(2), respectively. Significant difference in dendritic cell density was found among groups (F = 8.053, P = 0.000). When the injured area of the conjunctiva was limited, it was hard to discriminate the morphology of the conjunctival epitheliums in both Group A and Group B. Numerous inflammatory cells infiltrated in the conjunctival lamina propria and goblet cells were invisible. In Group C and Group D, the conjunctival epitheliums were almost normal. There were still some inflammatory cells and dendritic cells in the conjunctival lamina propria, and there were residual goblet cells visible in parts of the patients. When the injured area of the conjunctiva was large, the conjunctivas in four groups displayed hyperreflective stripe fibrous tissues instead of normal conjunctival epitheliums. CONCLUSIONS: The application of laser scanning confocal microscopy indicates that there is much difference on the cellular morphology of the ocular surface of severe alkali burns patients among diverse courses of the disease. The technique is a useful tool to the observation on the ocular surface of severe alkali burns patients.

[Advances in penetrating keratoplasty technique].

Rejection after corneal transplantation is currently the lead of the main reasons for corneal graft failure. Penetrating keratoplasty (PK) is still one of the gold standard surgeries for these patients. In recent years, improving the surgical techniques to reduce the incidence of corneal allograft rejection has been the way to more and more attention to the majority of ophthalmologists. To generalize and analyze the recent research of new technologies of femtosecond laser shaped penetrating keratoplasty of top-hat configuration (FS-TH-PKP), manual top-hat wound configuration for penetrating keratoplasty (M-TH-PKP), and half-top-hat penetrating keratoplasty (HTH-PKP) surgeries.

An in vivo confocal microscopy and impression cytology analysis of goblet cells in patients with chemical burns.

PURPOSE: To evaluate goblet cell density (GCD) on conjunctiva and cornea in patients with ocular chemical burns by in vivo laser scanning confocal microscopy (LSCM) and impression cytology (IC) and to explore the correlation between two methods. METHODS: Fifty-four patients (58 eyes) with chemical burn were enrolled in the study. LSCM was applied to identify the goblet cells on conjunctiva and cornea under in vivo conditions, and GCD was analyzed with the customized software. Impression cytology was then performed, and the biopsy specimens were stained to visualize goblet cells in vitro and to measure the density. Statistical software was used to analyze the correlation between GCD taken by two methods. RESULTS: Conjunctival goblet cells could be discriminated in 55 eyes and 57 eyes by in vivo LSCM and IC. They could be identified on the cornea in nine eyes and eight eyes by two methods. The positive rate of two methods had no significant difference. GCDs on conjunctiva measured by in vivo LSCM and IC were 136 +/- 79 cells/mm(2) and 121 +/- 66 cells/mm(2). Median GCDs on cornea detected by two methods were 30 cells/mm(2) and 23 cells/mm(2), respectively. A significant positive correlation was found between the GCDs on conjunctiva measured by these two methods as well as the GCDs on cornea. CONCLUSIONS: GCD decreased in patients with chemical burns. A positive correlation was found between GCD measured by in vivo LSCM and IC after chemical burns. In vivo LSCM was a promising device to study goblet cells in vivo under pathologic conditions.

Chromosome karyotype and Y chromosome microdeletion analysis in 133 idiopathic male infertile patients / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the relationship between chromosomal abnormalities and Y chromosome microdeletions in idiopathic male infertile patients.</p><p><b>METHODS</b>Karyotypic analysis and PCR were carried out in 133 male idiopathic infertile patients for chromosome karyotype and Y chromosome microdeletion analysis.</p><p><b>RESULTS</b>Of the 133 patients, 25 (18.80%) were found to have chromosome abnormalities, and 12 (9.02%) were found to have Y chromosome microdeletions including 2 with varicocele and 2 with cryptorchism. Four patients had both chromosomal abnormalities and Y chromosome microdeletions.</p><p><b>CONCLUSION</b>The chromosomal abnormalities and Y chromosome microdeletions may play an important role in idiopathic male infertility, suggesting the importance of examinations of chromosomal abnormalities and Y chromosome microdeletions in such patients.</p>

[Expression of regulatory T cells and related cytokines in mice allogenic corneal transplantation rejection].

OBJECTIVE: To study the changes of CD4(+)CD25(+) and CD8(+)CD103(+) regulatory T cells and their relevant cytokines in corneal allograft transplantation in mice. METHODS: BABL/c (H-2(d)) mice received corneal allografts from C3H/He (H-2(k)) mice were served as the experimental group. BABL/c (H-2(d)) mice received corneal from BABL/c (H-2(d)) mice were used as the control group. Corneal graft survival time was recorded pre-, 3rd day, 7th day, 4th week, 8th week post-operatively. Infiltration of inflammatory cells and corneal neovascularization were evaluated by histopathologic examination. The percentage of CD4(+)CD25(+) and CD8(+)CD103(+) T cell in peripheral blood and spleen was determined by flow cytometry. The expression of interleukin-10 (IL-10), IL-4, gamma-interferon (IFN-gamma) and IL-1beta in serum and aqueous humor was measured by enzyme-linked immunospecific assay (ELISA). RESULTS: The graft rejection in experimental group occurred at 7 days to 4 weeks after the operation, averaged (14.79 +/- 1.02) days. The grafts in the control group remained clear within 8 weeks observation and the survival time is much longer than that of the allografts (chi(2) = 46.934, P = 0.000). Flow cytometry showed that the percentage of CD4(+)CD25(+) T cell in peripheral blood in the control group after surgery was (3.36 +/- 0.29)%, (4.09 +/- 0.44)%, (5.44 +/- 0.35)%, (5.73 +/- 0.53)% at day 3, day 7, 4th week, and 8th week, respectively, which was significantly higher than that in the experimental group [(2.50 +/- 0.39)%, (3.24 +/- 0.25)%, (4.20 +/- 0.45)%, (4.18 +/- 0.14)%; t = 3.828, 2.898, 3.780, 4.892; P < 0.05]. On the other hand, CD8(+)CD103(+) T cell in peripheral blood in the experimental group after surgery was (2.20 +/- 0.33)%, (2.79 +/- 0.57)%, (4.55 +/- 1.03)%, (4.31 +/- 0.07)% at different periods, which was much higher than that in the control group (t = 7.133, 4.876, 5.196, 19.960; P < 0.05). The changes of CD4(+)CD25(+) and CD8(+)CD103(+) T cells in the spleen was earlier than those in peripheral blood. ELISA showed the expression of IL-10 and IL-4 in the serum in experimental group after surgery was much lower than that in the control group (t = 3.203, 3.141, 3.012, 2.869 and 2.340, 6.681, 8.839, 8.574; P = 0.011, 0.012, 0.013, 0.019 and 0.053, 0.000, 0.000, 0.000). The serum levels of IFN-gamma and IL-1beta in experimental group were higher than that in the control group (t = 3.508, 3.265, 4.402, 5.539 and 3.630, 5.796, 1.728, 0.660; P = 0.006, 0.011, 0.002, 0.000 and 0.005, 0.000, 0.115, 0.524). Furthermore, the cytokine levels in the aqueous humor behaved similarly with those in the serum. CONCLUSION: The down-regulation of CD4(+)CD25(+) Treg, IL-10 and IL-4 and the up-regulation of CD8(+)CD103(+) Treg, IFN-gamma and IL-1beta in mice allograft corneal rejection may play an important role in allograft rejection.

[Roles of angiopoietins family and its receptor in the pathogenesis and regulation of corneal neovascularization].

Corneal neovascularization is one of the tough problems in the cornea diseases, but its pathogenesis is still unclear. Binding of angiopoietins family and its receptor Tie2 set up the signal transducing system of Ang/Tie2, which plays the important role in the angiogenesis and regulation. Recently, the study of the signal transducing system of Ang/Tie2 has archived some progression in the corneal neovascularization. So this article reviewed the advances of related role of angiopoietins family and its receptor in the pathogenesis and regulation of corneal neovascularization including the structure and function of angiopoietins and its receptor Tie2.

Pediatric penetrating keratoplasty in Shanghai: a retrospective multiple centre study from 2003 to 2007.

BACKGROUND: With advances in pediatric keratoplasty over the last two decades, the success rate of surgery has increased remarkably. However, few epidemiological studies in this field have been performed in China. The present study investigated the indications and characteristics of pediatric penetrating keratoplasty in Shanghai. METHODS: All records of pediatric keratoplasty performed on 156 eyes in 149 children (< 14 years) at four ophthalmic units in Shanghai during a 5-year period (2003 - 2007) were used for this retrospective study. Patients were from the Eye, Ear, Nose & Throat Hospital of Fudan University, Shanghai No.10 People's Hospital, Shanghai No.1 People's Hospital, and Shanghai Peace Ophthalmology Hospital; and included mostly school-age children (97 boys and 59 girls). RESULTS: The median age at surgery was 9 years with an interval quartile range of 6-12 years. Scarring after keratitis (29.5%, 46/156) and traumatic corneal scar (19.2%, 30/156) were the most common indications. Best Corrected Snellen visual acuity (BCSVA) was reported only in 72% (112/156) of cases. Visual acuity outcomes were significantly better for keratoplasty 1 year postoperatively compared to preoperative visual acuity (P = 0.001). Of all patients, 13% (14/112) achieved a BCSVA of 6/18 or better. None of the indications was associated with a higher rate of failure compared to after 1 year follow-up. CONCLUSION: We obtained valuable information on pediatric keratoplasty in Shanghai. There was no significant difference in graft survival rate among the two indications. Vision outcomes after corneal transplantation in Chinese children were significantly better compared with pre-operation status. In conclusion, corneal graft survival can be achieved in childhood.

[The influence of lens and vitreous resection on anterior chamber associated immune deviation in mice model].

OBJECTIVE: To explore the impact of lens and vitreous resection on the induction of the anterior chamber associated immune deviation (ACAID) in the mice, and its relationship with the time duration postoperatively. METHODS: The current study was an experimental research. Fifty-one Wistar mice were chosen to involve this study. After the surgeries, 50 Wistar mice were randomly divided into 5 groups. Other 5 SD mice and 1 Wistar mouse were used as the providers of epithelium removed corneal grafts to induce ACAID. Group A: corneal grafts from wistar mouse were implanted into the anterior chamber 1 week after the lens and vitreous resection. Group B: the spleen cells were injected into the nape of neck 1 week after the lens and vitreous resection. The corneal grafts from SD mice were implanted into the anterior chamber 1 week, 4 week and 8 week after the lens and vitreous resection in group C, D and E respectively. To induce the delayed type hypersensitivity (DTH), spleen cells were injected into the right ear pinna 1 week after the neck injection in group B, 2 weeks after the corneal grafts anterior chamber implantation in other groups. At the same time point as the DTH inducement, aqueous humor was collected to measure the concentration of transforming growth factor (TGF) -beta2 and interleukin (IL)-10; heart blood was collected to detect the concentration of IL-4 and IL-10. And the spleens were removed to evaluate the expression of the GATA-3 mRNA by RT-PCR. The eyeballs were enucleated to evaluate the histopathologic changes. RESULTS: (1) The evaluation of DTH. The DTH were found in group B, C, D and E. (2) The analysis of the TGF-beta2 and IL-10 concentrations in aqueous humor and the IL-4, IL-10 concentrations in serum. The concentrations of IL-4 in serum were (4.073 +/- 0.198) ng/L in group A, (5.806 +/- 0.635) ng/L in group B, (5.535 +/- 0.278) ng/L in group C, (4.102 +/- 0.344) ng/L in group D and (5.313 +/- 0.317) ng/L in group E. The concentrations of IL-10 in serum in group A to group E were (7.854 +/- 2.349) ng/L, (25.633 +/- 6.307) ng/L, (40.103 +/- 16.010) ng/L, (14.321 +/- 2.983) ng/L and (28.620 +/- 5.251) ng/L. The IL-10 concentrations in the aqueous humor were (8.857 +/- 0.401) ng/L, (22.882 +/- 3.315) ng/L, (21.548 +/- 0.477) ng/L, (7.742 +/- 0.952) ng/L and (12.119 +/- 0.477) ng/L, respectively. The concentrations of TGF-beta in the aqueous humor were (5.800 +/- 2.899) ng/L in group A, (60.010 +/- 0.000) ng/L in group B, (57.055 +/- 4.179) ng/L in group C, (28.490 +/- 4.144) ng/L in group D and (36.370 +/- 3.169) ng/L in group E. Significant differences were found in concentrations of TGF-beta2 and IL-10 in aqueous humor and the concentrations of IL-4 and IL-10 in serum in group C, D and E. The concentrations of TGF-beta2, IL-4 and IL-10 were significantly lower in group D than those in group C and E. The trend that the concentrations of TGF-beta2, IL-4 and IL-10 increased after one week postoperatively, then decreased, and finally increased again was observed. (3) The expression of GATA-3 mRNA in spleen. The expression values of GATA-3 mRNA were 662.5 +/- 114.4 in group A, 730.7 +/- 53.8 in group B, 881.9 +/- 10.7 in group C, 1288.3 +/- 258.0 in group D and 1129.7 +/- 95.7 in group E. An obvious up-regulation of the GATA-3 mRNA was noted in group D and E. CONCLUSIONS: The eye after the lens and vitreous resection lost the ability to induce ACAID in the research duration of this study. But with the gradually decrease of the intraocular inflammation, the ability to induce ACAID might recover with the time.

[Variations of the microstructure in human limbus by confocal microscopy in vivo].

OBJECTIVE: To demonstrate the variations in the niche of limbal stem cells and the limbal basal epithelial cells in vivo by confocal microscopy. METHODS: One hundred and twenty eyes of 120 healthy subjects were enrolled in this study after a routine slit-lamp examination. They were divided into four groups according to the age: group A (0-), group B (20-), group C (40-) and group D (60 - 79). Confocal microscopic examination in vivo was performed and the images of the inferior limbus were recorded. The morphologic features of each group were analyzed and the average size of limbal basal epithelial cells of each group were measured. RESULTS: The morphologic features of limbus varied among groups. 96.7% (29/30) of group A showed clear Vogt Palisades, while 3.3% (1/30) showed no Vogt Palisades. 96.7% (29/30) of group B also showed clear Vogt Palisades, while 3.3% (1/30) showed no Vogt Palisades.70.0% (21/30) of group C showed clear Vogt Palisades, while 13.3% (4/30) showed atrophic Vogt Palisades and 16.7% (5/30) showed no Vogt Palisades at all. 33.3% (10/30) of group C showed clear Vogt Palisades, while 10.0% (3/30) showed atrophic Vogt Palisades and 56.7% (17/30) showed no Vogt Palisades at all. The average size of limbal basal epithelial cells in each group was (9.89 +/- 1.12), (10.65 +/- 1.45), (10.70 +/- 1.39) and (12.22 +/- 1.42) microm in sequence. CONCLUSIONS: There is variations in the microenvironment of niche in human limbus. Meanwhile, limbal basal epithelial cells showed different quantity and cell size, demonstrating varied proliferative potential.

[Comparative study on deep lamellar keratoplasty and penetrating keratoplasty for keratoconus].

OBJECTIVE: To investigate the therapeutic outcomes of deep lamellar keratoplasty (DLKP) and penetrating keratoplasty (PKP) for keratoconus. METHODS: Retrospective review of 29 eyes of 29 keratoconus patients, who underwent surgery in Eye & ENT Hospital of Fudan University from April in 2003 to April in 2006. Eleven eyes underwent DLKP and the rest had PKP. Pre- and post-operative visual acuity, astigmatism, complications and post-operative graft status were assessed. RESULTS: 82% of patients (9 eyes) in DLKP group gained BCVA better than 0.5, while in PKP group the proportion was just 78% (14 eyes). Postoperative astigmatism of two groups had no statistical significant difference with the value (-4.03 +/- 1.87) D and (-3.43 +/- 2.31) D respectively (DS: t = 2.135, P = 0.460, DC: t = -0.643, P = 0.528). The confocal image of epithelial cell, basal epithelium and Bowman's membrane was similar to that of normal cornea. The stromal cell was a little bit smaller and disordered. The endothelium in DLKP-treated eyes had normal cellular size and shape. There was no statistical significance between the density of operated eyes and contralateral unoperated eyes (2311.72 +/- 439.73) cells/mm2 and (2477.81 +/- 535.92) cells/mm2 respectively (t = 1.060, P = 0.780). However, the endothelial cells in PKP-treated eyes were highly pleomorphic with a decreased cellular density of (1642.17 +/- 583.41) cells/mm2, whereas the contralateral unoperated eyes had endothelium density of (2739.05 +/- 401.77) cells/mm2. The difference was statistical significance (Z = 7.32, P = 0.006). Complication rates were similar for DLKP and PKP, although the classification of the complications varied, being less severe in the DLKP group. CONCLUSIONS: DLKP seems to be a safe alternative for patients with keratoconus because of its equivalent effect to PKP. DLKP is more technically challenging but allows the risk of endothelial rejection to be avoided and may reduce the risk of late endothelial failure.

[Clinical analysis of keratoplasty for 121 children in Shanghai].

OBJECTIVE: To determinate the clinical characteristics of children underwent corneal transplantation in Shanghai, including the indications, the age and gender distributions, etc.. METHODS: Records of 122 keratoplasties in 122 eyes of 121 children below 17 years old, undertaken from January 1, 2003 to January 1, 2006, were gathered by four ophthalmic units in Shanghai as a multiple centre study and the data were analyzed by the statistic software SPSS 13.0 retrospectively, especially on the indication, the age and gender distributions. RESULTS: One hundred and twenty one patients were enrolled in this study. Traumatic corneal scar (22.3%) and scarring after keratitis (19.8%) were the leading indications for keratoplasty, followed by congenital corneal opacity (11.6%), keratoconus (10.7%), corneal ulceration (9.1%), graft rejection (5.0%), band keratopathy (5.0%), chemical injury (5.0%), thermal burns (4.1%), corneoconjunctival dermatosis (4.1%) and congenital hereditary corneal endothelial dysfunction (3.3%). The median age was 11.0 years with the interval quartile range at 8 - 14 years. The number of male patients (77) was approximately twice as many as the female patients (44). Ninety-five patients underwent penetrating keratoplasty (78.5%), 19 patients underwent lamellar keratoplasty (15.7%) and 7 patients underwent combined penetrating keratoplasty with other surgeries (5.8%). CONCLUSIONS: Traumatic corneal scar and scarring after keratitis were the leading indications. The age of operated cases mainly in the school age. Male is more frequent than the female.