Rhabdomyoma of the orbit.

The authors describe a rare case of orbital rhabdomyoma in a 3-year-old girl who presented with progressive proptosis of the left eye. An axial computed tomographic scan of the left orbit demonstrated an irregular retrobulbar mass. The tumor was resected locally from the lateral wall of the orbit and the resected specimens were diagnosed as orbital rhabdomyoma. The authors review the literature and discuss the diagnostic implications and treatment strategies.

[Histopathological and ultrastructural characteristics of oil-associated complications in silicone oil-filled human eyes].

OBJECTIVE: To observe morphological changes of oil-associated complications in silicone oil-filled human eyes, and to further explore their pathogenesis. METHODS: The morphology analysis and immunohistochemistrical study were performed in 25 specimens including 8 eyeballs, 1 ocular content, 4 preretinal membranes, 4 corneal buttons and 8 lens from human eyes with silicone oil tamponade. Two of preretinal membranes acquired freshly were also evaluated by transmission electron microscopy (TEM). RESULTS: Endothelium cell loss (90%) and band keratopathy (83%) were the most typical changes in silicone oil-associated keratopathy;while epithelial cell fibrosis was the most frequent histopathological features in silicone oil-associated cataract. In 8 eyeballs and 1 ocular content, it was found that damages to normal retinal layers and formation of preretinal or subretinal membrane with extensive silicone bubbles were obvious in the cases of silicone oil-associated retinopathy, which included loss and degeneration of neuron cells. Moreover, in 3 eyeballs with silicone oil for more than 60 months, retinas were completely replaced by fibril membranes, and the oil vacuoles were also found in sclerocorneal scar, trabecula, iris, ciliary body, choroid, optic nerve and its tunica vaginalis. These finding demonstrated that the longer the silicone oil was retained in eyeballs, the more severe its complications were. Different sizes of silicone bubbles in 2 preretinal membranes were noted easier by TEM than light microscopy. There were some macrophages marker (CD68) positive staining cells in the tissues filled with silicone bubbles, such as preretinal or subretinal membrane and optic nerve. Partial of the membranes surrounding the oil bubbles was positive for GFAP staining, and other part was positive stained for Vimentin. CONCLUSIONS: Intraocular silicone oil can damage the normal tissue structures and function if it is retained in eyeballs too long. This results suggest that silicone oil should be removed timely after the retinal reattachment stabilized and can not be used as a kind of long term intraocular tamponade.

[An experiment study of in vitro induced antitumor immune responses by vaccination of tumor lysate-pulsed dendritic cells to kill SO-RB(50)].

OBJECTIVE: To investigate induced antitumor immune responses by vaccination of tumor lysate-pulsed dendritic cells (DC) to kill retinoblastoma cells SO-RB(50). We hope to offer new approach for the treatment of patients with retinoblastoma. METHODS: DC was pulsed with RB tumor lysates in vitro and incubated with autologous lymphocytes to induce antigen specific CTL (cytotoxic T lymphocyte, CTL). SO-RB(50) cells were used as target cells and Raji cells were used as control target cells. Cytotoxicity of CTL was evaluated by MTT method (methyl thiazolyl letrazolium). The specific cytotoxicity of CTL to SO-RB(50) and Raji cells was compared. The cytotoxicity of CTL from RB and normal subjects was compared between these two groups. RESULTS: Antigen specific CTL showed greater cytotoxicity to SO-RB(50) than Raji cells, the difference was statistically significant, P < 0.01. The cytotoxicity was dose-dependent to the ratio of CTL/target cell. The nonspecific cytotoxicity to Raji cells was the same in CTL from RB patients and normal subjects, P > 0.05. The specific cytotoxicity of CTL from RB patients to SO-RB(50) was weaker than that from the healthy subjects, P < 0.01. CONCLUSION: DC pulsed with RB tumor lysate in vitro can induce antigen specific CTL which can kill the SO-RB(50) target cells specifically. This method may have potential value of therapy for the RB patients.

Angioleiomyoma of the ciliary body: a case report.

PURPOSE: To report a rare case of angioleiomyoma of the ciliary body METHODS: The clinical manifestation, imaging findings, histopathologic characteristics were analyzed in a 32-year-old male patient with angioleiomyoma of the ciliary body. RESULTS: The tumor was removed intact with local resection. Histopathologic examination revealed that the tumor was full of vessels and it was composed of spindle cells with abundant cytoplasm. Immunohistochemical studies showed positive for SMA and Desmin and negative for S100 and HMB-45. CONCLUSIONS: Angioleiomyoma of the ciliary body is a rare tumor that can be successfully treated with local surgical resection in this area. It needs to be differentiated from other tumors, especially malignant melanoma.

Experimental inhibition of corneal neovascularization by endostatin gene transfection in vivo.

OBJECTIVE: To investigate endostatin gene therapy of rat corneal neovascularization induced by acid cauterization. METHODS: pBlast-hEndostatin and pBlast-Mcs were identified by digestion with Nhe Iand Sal I, by PCR reaction, by sequence, and then by alignment of PCR products with the gene Bank using NCBIBLAST software. They were then purified with QIAGEN Endofree plasmid maxi kit. Rat corneal neovascularization models were made with 75% AgNO(3) and 25% KNO(3) cauterization. The treatment method was subconjunctive injection of the pBlast-hEndostatin with the control of pBlast-Mcs. RESULTS: pBlast-hEndostatin was found to contain the human endostatin gene. The rat corneal neovascularization induced by acid cauterization was significantly suppressed after subconjunctive injection of the pBlast-hEndostatin with inhibition rates of 37%, 40.2%, and 42.8% respectively on the sixth, tenth, and fifteenth day. The inhibition rate for the density of corneal neovascularization was 40%. However, no inhibition effect on the length of the neovascularization and corneal inflammatory cells was observed. Corneal neovascularization areas were positively correlated with edema and corneal opacity. CONCLUSIONS: The plasmid of pBlast-hEndostatin contained the human endostatin gene. The rat corneal neovascularization induced by acid cauterization can be partially inhibited by subconjunctive injection of the pBlast-hEndostatin mediated by liposomes. Endostatin produced by transfected fibroblast cells directly inhibits corneal neovascularization. This is not caused by inflammatory reaction inhibition.

[Clinical analysis of tumors of the iris and ciliary body].

OBJECTIVE: To evaluate the clinical features, different diagnosis, pathological features, management, and prognosis of tumors of the iris and ciliary body. METHODS: Medical records, photographs, pathological findings and the results of follow-up of 37 cases with tumors of the iris and ciliary body were reviewed as a retrospective study. RESULTS: Of the 37 cases with tumors of the iris and ciliary body, 26 were male and 11 were female. The mean age at diagnosis was 38 years, ranged from 5 - 64 years. According to the histopathological examination, melanoma was the most common tumor in the iris and ciliary body (15 cases, 40.5%), followed by metastatic tumors (8 cases, 21.6%), teratoid medulloepitheliomas (3 cases, 8.1%) and iris nevus (2 cases, 5.4%). MANAGEMENT: The tumors were excised in 14 cases. Enucleation was performed in 21 cases. Two cases were observed without any surgical treatment. Thirty-four cases were followed-up for 2 months to 15 years, averaged 31 months. Most melanomas of the iris and ciliary body are round or semi-spherical dark brown vascularized mass, with engorged episcleral sentinel vessels in some cases. The tumor showed a shadow during transillumination. Ultrasound biomicroscopy revealed a low to medium echoic solid lesion, with echoic changes in adjacent infiltrated tissues. Melanoma showed positive immunoreactivity for melanoma-specific antigen, and had a good prognosis. Metastatic tumors of the iris and ciliary body were flat or near round, dirty, single or multiple neoplasms, growth rapidly, with abundant neovascularization, and had a poor prognosis. Primary carcinomas could be found in other parts of the body. CONCLUSIONS: Melanoma of the iris and ciliary body has typical features that may help to distinguish them from other tumors. Metastatic tumor has characteristic features, but the diagnosis can be made only with supplementary examination and immunocytochemical studies. Medulloepitheliomas should be differentiated from retinoblastoma.

[Preparation of anti-human retinoblastoma monoclonal antibody by solid tumor cell immunization and study on characteristics of its antigen].

OBJECTIVE: To prepare anti-human retinoblastoma (Rb) monoclonal antibody (McAb) by immunization with solid tumor cells and preliminarily study the characteristics of its antigen. METHODS: Three Balb/C mice were immunized by intraperitoneal injection of Rb solid tumor cells acquired from enucleation. Spleen lymphocytes were separated from them and fused with myeloma cell line SP2/0. The anti-human Rb McAb was selected by Rb solid tumor cell, SO-Rb(50), SO-Rb(70), etc as antigens. The characteristics of its antigen were preliminarily studied by immunofluorescence, immunohistochemistry and Western blot. RESULTS: After repeatedly cloning them with micro-manipulation equipment, we successfully established 3 hybridoma cell lines secreting anti-human Rb McAb. Of all, 3C6 hybridoma cells could steadily secrete anti-human Rb McAb after they were frozen, resuscitated and passaged repeatedly for 2 years. Its subgroup was IgG(1). Both immunofluorescence and immunohistochemistry examinations demonstrated that the corresponding antigen of McAb 3C6 was specifically and highly expressed in Rb tumor cell membrane and cytoplasm. The other tumor tissues and human normal eye tissues were negative. Western blot analysis preliminarily demonstrated that McAb 3C6 could bind 25 000 protein band of Rb antigen. CONCLUSIONS: The anti-human Rb McAb 3C6 established by immunization of solid tumor cells is specifically and highly expressed in Rb tumor cells and has no cross-response with other tumor tissues and normal eyeballs. The antigen molecular weight bounded to this antibody is 25 000 or so, which further shows that it is a possibility for a new unidentified gene to be concerned with the tumor formation of Rb.

[An experimental study on effect of perfluorohexyloctane on corneal endothelial cells].

OBJECTIVE: To investigate the effect of perfluorohexyloctane on corneal endothelial cells in rabbit eyes. METHODS: Fifteen New Zealand albino rabbits were divided into two groups: experiment group (F(6)H(8)) and control group (BSS). All rabbits underwent anterior chamber injection of 0.15 ml F(6)H(8) or BSS. Slit-lamp biomicroscopy and corneal endothelium photography were performed pre- and post-injection. Histopathological examination and transmission electron microscopy (TEM) were performed after the rabbits were sacrificed. RESULTS: All the corneas were clear. Since 4 weeks postoperatively, the endothelial cells were markedly irregular in size and shape, and the number of endothelial cells was markedly decreased. Multilayered retrocorneal membranes grew gradually from 4 weeks after surgery. Vacuolar degeneration was seen in some endothelial cells. Nuclear degeneration and edema of endoplasmic reticulum were seen in TEM. CONCLUSION: Corneal endothelial cells degenerated after contacting with F(6)H(8) for 4 weeks. As a silicone solvent, F(6)H(8) should be removed out completely after injection. It is not recommended to apply this agent as a new intraocular temponade.

Pluripotent embryonic stem cells developed into medulloepithelioma in nude mice eyes.

PURPOSE: The pluripotent embryonic stem cells can differentiate into various kinds of normal tissues. There is no previous report on the differentiation of embryonic stem cell in the intraocular environment. In this paper, the authors tried to investigate the intraocular growth character of mice embryonic stem cells in nude mice. METHODS: Murine embryonic stem cells were cultured and maintained in an undifferentiated state in vitro. They were transplanted into the right eyes of 20 nude mice by microinjection under operating microscope. Animal eye observation, light microscope and immunohistochemical examinations were implemented. RESULTS: Two to three days after transplantation, small pieces of gray-white material could be viewed in the vitreous cavity. Between the 15th and 20th day, the gray-white mass grew into the anterior chamber in 4 nude mice eyes. Then, the mass at the anterior chamber extended extraocularly. On the 30th day, a remarkable proptosis was observed in two of the four nude mice. In 6 to 45 days, the mice were executed for morphological examination which showed the following typical structures: (1) Undifferentiated cells with prominent nucleolius. (2) Flexner-Wintersteiner-like rosettes. (3) Medulloepithelioma-like structure: the cells were arranged in sheets, cords, tubes, and cysts. (4) Large, spindle-or astrocyte-like cells. (5) Cartilage-like structure. Immunohistochemically, most of the cells were highly positive in NSE staining and a few cells were moderately positive in GFAP staining. CONCLUSIONS: Both animal eye findings and morphologic examinations certificated that the transplanted embryonic stem cells could grow in the eyes of nude mice and differentiate into intraocular medulloepithelioma.

Experimental study of effect of perfluorohexyloctane to retina.

OBJECTIVE: To investigate the effect of perfluorohexyloctane to the retina of rabbit eyes. METHODS: Perfluorohexyloctane (experimental group) or BSS(control group) into vitreous cavities of fifteen vitrectomized New Zealand white rabbits. A slit-lamp biomicroscope and an indirect ophthalmoscope were used to examine all the eyes pre- and postoperation. Histopathological examination was performed after the rabbits were sacrificed. RESULTS: Perfluorohexyloctane was injected into the vitreous cavity forming a single large layerclear globe. No retinal detachment and cataract were found. The edema of outer plexiform was significant, and then get a darker cytoplasm and nucleoplasm. CONCLUSIONS: Perfluorohexyloctane in vitreous cavity had significant side effects on retina. As a silicone solvent, it should be removed out completely after injection. We don't recommend to use it as a new intraocular tamponade.

Cloning and sequence analysis of light variable region gene of anti-human retinoblastoma monoclonal antibody.

PURPOSE: To clone the variable region gene of light chain of monoclonal antibody against human retinoblastoma and to analyze the characterization of its nucleotide sequence as well as amino acid sequence. METHODS: Total RNA was extracted from 3C6 hybridoma cells secreting specific monoclonal antibody (McAb) against human retinoblastoma (RB), then transcripted reversely into cDNA with olig-dT primers. The variable region of the light chain (VL) gene fragments was amplified using polymerase chain reaction(PCR) and further cloned into pGEM -T Easy vector. Then, 3C6 VL cDNA was sequenced by Sanger's method. Homologous analysis was done by NCBI BLAST. RESULTS: The complete nucleotide sequence of 3C6 VL cDNA consisted of 321 bp encoding 107 amino acid residues, containing four workframe regions (FRs) and three complementarity-determining regions(CDRs) as well as the typical structure of two cys residues. The sequence is most homological to a member of the Vk9 gene family, and its chain utilizes the Jkl gene segment. CONCLUSION: The light chain variable region gene of the McAb against human RB was amplified successfully, which belongs to the Vk9 gene family and utilizes Vk-Jkl gene rearrangement. This study lays a good basis for constructing a recombinant antibody and for making a new targeted therapeutic agents against retinoblastoma.

[The study of immunofluorescence on the retinal digest preparations].

PURPOSE: To study the technique of immunofluorescence staining on the digested retina preparations. METHODS: Retinas of 5 normal Wistar rats and 5 STZ diabetic Wistar rats of were used to make digested retina preparations and carry out immunofluorescence staining with the vascular cell adhesion molecule-1 (VCAM-1). RESULT: The retinal vessels were clearly observed; There were a lot of positive cells in the retinal preparations of the diabetic Wistar rats, while there were no positive cells in the retinal preparations of the normal rats. CONCLUSION: It is feasible to carry out immunofluorescence staining on the digested retina preparations.