Objective To evaluate the effectiveness and safety of mushroom incision femtosecond-assisted penetrating comeal transplantation combined cataract extraction surgery.Methods A retrospective analysis was carried out for 29 patients (30 eyes) who performed FS-PKP combined cataract extracapsular extraction versus 11 patients (11 eyes) who performed traditional PKP combined cataract extracapsular extraction.Lesions involving whole comea and all the patients' lenticular opacity were moderate or above.Femtosecond laser was used for make mushroom incision of comeal graft and corresponding recipient bed,conventional group used trephine.Then implemented cataract extracapsular extraction,intraocular lens were implanted,sutured at last.Their follow-up were 1 week,1 month,3 months and 6 months,part of the patient were followed up until 18 months.The follow-up results of two groups were compared.Results Postoperative vision of two groups had increased,postoperative UCVA and BCVA of femtosecond group were better than normal group,postoperative astigmatism was lower than normal group within 3 months,and there was no obvious difference after 6 months.No obvious difference was found between two groups of ECD and postoperative complications occurrence rate.Conclusions Mushroom incision of femtosecond-assisted penetrating corneal transplantation combined cataract extraction surgery has obvious curative effect,precision cutting and high security,its early visual acuity is superior to the traditional penetrating keratoplasty,it can be widely used in clinical therapy.

AIM To design, optimize and validate a rapid, internally controlled real-time polymerase chain reaction (RT-PCR) test for herpes simplex virus (HSV) in the diagnosis of necrotizing herpes stromal keratitis. METHODS Tears alone or together with corneal epithelium scrapings from 30 patients (30 eyes) suspected of necrotizing herpes stromal keratitis were tested for HSV DNA by RT-PCR. The samples were collected during the first visit and then on the subsequent 7, 14, 28, 42, and 56d. The symptoms of the patients were scored before treatment to determine the correlation between HSV concentration in the corneal epithelium scrapings and clinical scores. RESULTS The positive rate (46.4%) in the corneal epithelium group before the therapy was significantly higher than that (13.3%) in the tears group (P=0.006). There were 13 positive HSV patients before the therapy, the concentration of HSV DNA in corneal epithelium scrapings group was significantly higher than that in the tears group (paired t-test, P=0.0397). Multilevel mixed-effects model analysis showed that the difference between the corneal epithelium scrapings group and the tears group was statistically significant (P=0.0049). The Spearman rank correlation analysis indicated a positive correlation between the HSV concentration in the corneal epithelium scrapings and clinical scores before the treatment (r=0.844, P<0.0001). CONCLUSION RT-PCR appears to be a powerful molecular tool for the diagnosis of necrotizing herpes stromal keratitis.

Background Herpes stromal keratitis (HSK) is a common infectious ocular surface disease,with a higher recurrent rate,especially in necrotizing HSK.The diagnosis of HSK primarily depends on signs and symptoms,and specific laboratory diagnostic is lack.Objective This study was to clarify the expression of herpes simplex virus (HSV) in the corneal epithelium scrapings and tears of necrotizing HSK patients.Methods Thirty eyes of 30 patients with necrotizing HSK were enrolled in Nanjing Hospital of Nanjing Medical Hospital from September 2012 to September 2013 under the patient's informed consent.The eyes were examined by slit lamp microscope and scored.HSK patients received local and systemic therapy for 8 weeks and then an oral maintenance dose for 6 months.Corneal epithelial scrapings and tears samples were collected for HSV DNA detection by real-time PCR before and the 1 st,2nd,4th,6th and 8th week after therapy respectively.The difference of HSV positive rate was compared between corneal epithelium scrapings and tears samples using Chi-square test.Multilevel mixed effective model was employed to evaluate HSV concentration change in the samples at various time points in the HSV-positive patients of initial visit.The correlation between HSV concentration and clinical score was analyzed by Spearman rank correlation.Results HSV-positive rate was 46.4％ (13/30) in the corneal epithelial scrapings and 13.3％(4/30) in the tear samples,showing a significant difference between them (P =0.006).HSV-positive rate was significantly lower in the corneal epithelial scrapings 1 week,2 weeks and 4 weeks after treatment than before (P =0.001,0.003,0.004),and no HSV was detected 6 weeks and 8 weeks after treatment.No significant change in HSV-positive rate in tear samples in 1 week and 2 weeks after treatment in comparison with before treatment (P =1.000,0.583),and no HSV was detected after 4 weeks following treatment.The HSV concentration was 2460 (2 165-636500)/ml in initial 13 HSV-positive eyes of corneal epithelial scrapings and 0 (0-1150)/ml in initial 13 HSV-positive eyes of tear samples.Multilevel mixed effective model determined that HSV concentration was significantly lower in corneal epithelial scraping than that in tear (P =0.005),and HSV concentration was reduced with the lapse of time (P =0.001),with the faster rate of decline in the corneal epithelial scrapings (P =0.049).A positive correlation was found between initial HSV concentration and clinical scores (rs =0.844,P =0.000).Conclusiors Real-time PCR appears to be a powerful molecular tool for the detection of HSV in the HSK,especially in corneal epithelial scrapings of lesion.The initial positive outcome of viral DNA in corneal epithelial scrapings predicts a severe clinical procedure.

Femtosecond laser has the characteristic of high precision cutting, so it has great application potential in keratoplasty.It can replace the mechanical microkeratome used for penetrating keratoplasty, lamellar keratoplasty and endothelial keratoplasty according to the clinical need to produce different shapes of corneal graft and the graft bed. Femtosecond laser provides a new way for improving and optimizing the operation of various keratoplasty. In this paper, we reviewed the use of femtosecond Iaser in keratoplasty.

AlM: To evaluate the therapeutic effects and safety of femtosecond laser assisted penetrating keratoplasty ( FS-PKP) . METHODS: ln the retrospective case series study, 46 cases ( 48 eyes ) with FS- PKP holds were analyzed. Preoperative best corrected visual acuity ( BCVA) was 2. 35 ± 1.01 (with a standard logarithmic eye chart). Opacities were all from epithelium to endothelium, including keratoconus, keratolukoma and corneal scarring etc. The full-thickness corneal donor-recipient grafts in different diameter and incision kerfangle were made by femtosecond laser in keratoplasty. The patients were treated according to the conventional penetrating keratoplasty after surgery. Evaluation indexes included preoperative and postoperative 1, 6, 10mo uncorrected visual acuity (UCVA), BCVA, corneal astigmatism (CA) and surgical complications. Forty-three cases (44 eyes) with traditional PKP were as control group. The data of two groups were analyzed by mean comparison of two groups and Chi-square test in SPSS 18. 0. RESULTS: All the operations were completed successfully, postoperative visual acuity of all patients had different degrees of increase. Comparison with the postoperative UCVA of traditional PKP surgery, UCVA was 3. 48 ± 0. 43 and 2. 97 ± 1. 14 at 1mo after operation, respectively, with statistically significant difference ( P=0. 009). The UCVA was 3. 86±0. 25 and 3. 74±0. 73 at 3mo after operation, respectively; the UCVA was 4. 04 ± 0. 23 and 3. 98 ± 0. 29 at 6mo after operation, respectively, with no statistically significant difference (P=0. 146, P=0. 56). ln FS-PKP group, BCVA was better than traditional PKP surgery group, with statistically significant difference ( P=0. 002 ). While there had no statistically significant difference between two groups of BCVA at 6, 10mo after operation (P6mo =0. 132, P10mo =0. 47). The incidence of complication of FS-PKP surgery, such as postoperative astigmatism, postoperative intraocular pressure and postoperative incision dehiscence were significantly lower than the one of traditional PKP surgery(P<0. 05). CONCLUSlON:Compared with traditional penetrating keratoplasty, FS-PKP provides significantly faster visual recovery in the early postoperative period, and induces significantly less corneal astigmatism and lower incidence of complication. lt reflected the clinical superiority of this new technology .

Herpes simplex keratitis( HSK) is the leading cause of infectious blindness. In general, diagnosis of HSK is based on clinical findings, which often are not confirmatory. With the development of molecular biology,a variety of laboratory methods that helped in achieving a diagnosis including in vivo confocal microscopy( IVCM),loop- mediated isothermal amplification( LAMP),real-time polymerase chain reaction( rt- PCR),nested PCR and multiplex PCR. They are considered as accurate and fast diagnostic techniques,thus characteristics of these are reviewed in this paper.