Cryopreserved limbal lamellar keratoplasty for peripheral corneal and limbal reconstruction.

This study aimed to evaluate the outcomes and described the recovery process of cryopreserved limbal lamellar keratoplasty (CLLK) for peripheral corneal and limbal diseases. Thirteen eyes of 12 patients with a mean age of 41±23.9y were included. The average follow-up was 12.1±5.6mo. Stable ocular surface was achieved in all eyes at last follow-up. Epithelialization originated from both recipient and graft in 9 eyes. We conclude that CLLK compensates for the shortage of donor corneas and cryopreserved limbal grafts provide epithelialization sources in ocular surface reconstruction.

Amniotic membrane transplantation with topical interferon alfa-2b after excision of ocular surface squamous neoplasia.

To evaluate the outcome of amniotic membrane transplantation (AMT) after tumor excision followed by topical interferon alfa-2b (IFNα2b) drops for primary ocular surface squamous neoplasia (OSSN). Twelve eyes of 12 patients with a mean age of 66±10y were included. The average follow-up was 23±10mo. All 12 patients had limbal involvement. Smooth ocular surface and transparent cornea were achieved in all cases. No sign of inflammation, neovascularization, symblepharon or recurrence was noted at the last follow-up. We conclude that AMT with topical IFNα2b drops restores a healthy ocular surface in OSSN without recurrence.

[Effect on the proliferation of bovine corneal endothelial cells by small hairpin RNA interference targeting p27Kip1].

OBJECTIVE: To clarify the proliferation of bovine corneal endothelial cells (bCEC) by interference with the recombinant plasmid of short hairpin RNA (shRNA) against p27Kip1, a kind of cyclin-dependent kinase inhibitor (CKI). METHODS: It was an experimental study. Three p27Kip1-shRNA template DNA sequences containing small hairpin structure were designed and synthesized as experimental groups. Plasmid expressing irrelevant shRNA with a random combination was used as negative shRNA. The products were inserted into the Pgensil-1 plasmid and the recombinant plasmid of Pgenesil-P1, Pgenesil-P2, Pgenesil-P3 and Pgenesil-HK were constructed. The recombinant plasmids were transfected into bCEC cells with liposome and a blank group. The expression of mRNA and protein of p27Kip1 was detected by RT-PCR and Western blot after stable transfection, and the plasmid with the best inhibitory effect was selected. The growth of the experimental group, Pgenesil-HK group and blank group were assessed by MTT. The influence of shRNA-p27Kip1 on bCEC cell cycle was deteceted by flow cytometry (FCM). All statistical analyses were performed using one-way ANOVA. RESULTS: Restrictive enzyme digestion and sequence analysis showed that four recombinant plamids were constructed successfully and the aim sequence was obtained. The expression of p27Kip1 mRNA and p27Kip1 protein of Pgenesil-P1 group, Pgenesil-P2 group and Pgenesil-P3 group were all lower than that in the control group, including blank group and negative siRNA group. The inhibitive rate of mRNA reached 32.71%, 67.76% and 80.28% (F = 453.102, P = 0.000 in each group) and the inhibitive rate of protein reached 29.27%, 64.73% and 76.13% (F = 75.385, P = 0.000 in each group) compared with the blank group. As the lowest expression among the three positive shRNA group, Pgenesil-P3 was selected for the next steps. There was no significant difference between blank group and negative Pgenesil-HK of the expression of p27Kip1 protein (P = 0.356) and the express of p27Kip1 mRNA (P = 0.246). Compared with the control group and the blank group, the growth of the bCEC transfected by Pgenesil-P3 was significantly promoted with increased cell percent of S-phrase (F = 334.957, P = 0.000) and decreased cell percent of G1-phrase (F = 134.224, P = 0.000). CONCLUSIONS: shRNA-p27Kip1 can down-regulate the expression of bCEC effectively and increase the growth of bCEC. shRNA-p27Kip1 RNA interference may be an effective method to promote the proliferation of CEC.

[Preliminary report of surgical treatment of vernal keratoconjunctivitis with giant papillae].

OBJECTIVE: To evaluate the efficacy and safety of excision and cryotherapy combine with amniotic membrane transplantation (AMT) for the treatment of vernal keratoconjunctivitis (VKC) with giant papillae (GP). METHODS: Eight patients (16 eyes) with VKC, characterized by GP on the upper tarsal conjunctiva, refractory to medical treatment, underwent excision and cryotherapy associated with AMT. The follow up period ranged 3-22 months. RESULTS: Corneal shield ulcers and superficial punctuate keratitis healed during the first week after the surgery and did not recur. Fourteen eyes were symptom free and without GP one month after surgery. No ectropion, trichiasis and other complications were noted. The blood vessels of upper tarsal conjunctiva could not be seen clearly, a small amount of conjunctival scar was observed. Recurrence of GP was observed in 2 eyes, with reduced area and lessened irritation symptom as compared with preoperative status. Among them, one eye was treated by cyclosporine eyedrops with improvement. The other eye did not improve, and underwent secondary surgery (a cotton patch soaked in fluorouracil was applied on the upper eyelid after excision and cryotherapy). Nine months after the treatment, the patient had no symptoms and GP did not recur. CONCLUSIONS: Excision and cryotherapy combine with AMT may be an effective and safe method for the treatment of refractory VKC with GP. But the candidates for surgical treatment should be chosen carefully.