ABSTRACT
OBJECTIVE: To clone the sequence of mutation type GJA8 gene (mGJA8) and wild type GJA8 gene (wGJA8) of a congenital inherited nuclear cataract family and study their expression in eukaryotic cell lines in vitro. METHODS: The mGJA8 and wGJA8 were amplified from this family's DNA and healthy people's DNA by PCR respectively. The mGJA8 and wGJA8 were recombined with plasmid pEGFP-N1 respectively. The accuracy of pEGFP-N1-GJA8 was confirmed by restriction enzyme digestion and DNA sequencing. Finally pEGFP-N1- mGJA8 and pEGFP-N1- wGJA8 and GFP protein were transfected into COS7 cells by lipofectin. The expression of pEGFP-N1-GJA8 and GFP fusion protein were to observe under fluorescence microscope, and to detect by Western-blotting and immunohistochemical staining. RESULTS: The mGJA8 and wGJA8 were cloned successfully. With restricting enzyme digestion analysis and DNA sequencing, recombinant plasmid pEGFP-N1-mGJA8 and pEGFP-N1-wGJA8 were constructed correctly and their GFP fusions were expressed in transfected COS7 cells. The expression of pEGFP-N1-mGJA8 and pEGFP-N1-wGJA8 fusion protein were observed under fluorescence microscope, and detected by Western-blotting and immunohistochemical staining successfully. CONCLUSIONS: The mGJA8 gene and wGJA8 gene are cloned successfully, and pEGFP-N1-mGJA8 and pEGFP-N1-mGJA8 fusion protein can be expressed in COS7 cells, which establish the foundation for further studying the mechanism of this congenital inherited nuclear cataract family.
Subject(s)
Connexins/biosynthesis , Connexins/genetics , Eye Proteins/biosynthesis , Eye Proteins/genetics , Animals , COS Cells , Chlorocebus aethiops , Cloning, Molecular , Genetic Therapy , Genotype , Humans , Mutation , Nuclear Family , Pedigree , Plasmids , Recombinant Fusion Proteins/biosynthesis , Recombinant Fusion Proteins/genetics , TransfectionABSTRACT
AIM: To describe the role of endoscopic transnasal canaliculorhinostomy (ETC) in refractory common canalicular obstruction (CCO) associated with an absent or unidentifiable lacrimal sac. METHODS: The records of patients with refractory CCO who underwent ETC at the Eye Hospital of Wenzhou Medical University from October 2007 to December 2016 were retrospectively reviewed. RESULTS: Fifty-six patients (56 eyes) with refractory CCO were recruited into the study. Eight patients were excluded due to the presence of a residual lacrimal sac or failure to complete the follow-up duration. The anatomic and functional success rates were both 85.4% (41/48) at a mean follow-up of 18.6mo. Five cases failed as a result of ostial synechia and two failed because of ostial obstruction by granulation. Postoperative complications included mild nasal bleeding in 5 cases, dried nasal feeling in 8 cases, and olfactory dysfunction in 4 cases. CONCLUSION: Although being surgically challenging, ETC has comparable findings to its external approach counterpart or conjunctivodacryocystorhinostomy (CDCR) with Jones tube. And it may prove to be a novel alternate surgical technique for patients with refractory CCO without identifiable lacrimal sac.
ABSTRACT
To explore the effect of high glucose concentration on the expression of vascular endothelial growth factor (VEGF) and pigment epithelium derived factor (PEDF) in the cultured rat retinal Müller cells. Rat Müller cells were cultured and RT-PCR and Western-blot analysis were used to measure the levels of VEGF and PEDF in cultured Müller cells at different high glucose concentrations. Under 10, 20, 30 mmol/L high glucose conditions, the levels of VEGF mRNA and protein increased and the levels of PEDF mRNA and protein decreased. These results suggest that the VEGF and PEDF expression in Müller cells are unbalance under high glucose concentration, which contribute to retinal neovascularization in diabetic retinopathy.
Subject(s)
Eye Proteins/biosynthesis , Glucose/pharmacology , Nerve Growth Factors/biosynthesis , Retina/drug effects , Serpins/biosynthesis , Vascular Endothelial Growth Factor A/biosynthesis , Animals , Cells, Cultured , Eye Proteins/genetics , Gene Expression/drug effects , Immunohistochemistry , Nerve Growth Factors/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Retina/cytology , Retina/metabolism , Serpins/genetics , Vascular Endothelial Growth Factor A/geneticsABSTRACT
RATIONALE: Though giant malignant tumors arising in the eyelid are rare, they often require extensive surgery for removal along with orbital exenteration. Because of this, repairing orbital defects is an important factor in the surgical strategy. PATIENT CONCERNS: Case 1 was a 78-year-old nomad man who presented in the Department of Ophthalmology with a giant tumor in his right eyelid, which had developed over three years. Clinical examination revealed a huge pigmented, nonhealing ulcerated lesion, approximately 52×44×40âmm in size. Case 2 was a 52-year old rural male complaining of a huge tumor in the right eyelid. Patient medical history revealed that the mass was initially the size of a soybean and gradually grew over 3 years to the size of a fist. DIAGNOSES: Histopathological examination of the tumors revealed that one was a basal cell carcinoma and the other a sebaceous gland carcinoma. INTERVENTIONS: The two cases of giant malignant eyelid tumors were surgically excised using rapid frozen section margin control. Different pedicle myocutaneous flaps were used to repair the orbital skin defects. OUTCOMES: Postoperative follow-up showed perfect healing of the pedicle flaps and good patient compliance. The results of these cases indicate that covering exposed orbital cavities with composite pedicle mycuaneous flaps is a simple and practical strategy for orbital reconstruction. Not only does this help maintain orbital stability, but it also provides opportunities for patients to return to normal lives. LESSONS: Although surgical management is often the first option for treatment of giant eyelid tumors, recurrence and mortality due to the tumors is still high after long-term follow-up. Therefore, early discovery and treatment is the best way to control the progression of giant eyelid tumors and enhance survivability.