The amniotic membrane has been shown to have anti-inflammatory,anti-fibrotic,anti-angiogenic properties and ability to provide a substrate for the growth of corneal and conjunctival epithelial cells,and it is an ideal material for ocular surface reconstruction.The treating patten of corneal and ocular surface diseases has changed due to the widespread using of amniotic membrane transplantation,but there are a lot of problems in the application of amniotic memebrane in clinic,such as how to grasp indications and reduce the failure rate,how to apply individual skill for different patients with corneal disease,and how to observe postoperative complicationsm and reasonably use medicine after operation.This paper give some personal experience and opinion in orde to achieve better effects in treatment of corneal and ocular surface diseases using amniotic membrane transplantation.

Background In recent years,incidence of drug-induced keratopathy is increasing highly.Druginduced keratopathy is lack of typical clinical features and offen confused with the primary disease.Therefore,summarizing and concluding the clinicals feature and standard treatments of drug-induced keratopathy are key problem need to be solved urgently for us.Objective This study was to retrospectively analyze the clinical features and therapeutic procedure of drug-induced keratopathy.Methods A retrospective case series analysis method was adopted.The clinical data of 36 eyes (31 patients) with drug-induced keratopathy were collected by Shandong Eye Hospital from 2008 to 2012,including eye disease history,medication history,medication dosage and duration.A series of relevant examinations were performed,including best corrected visual acuity (BCVA) before and 1 month after treatment,Schirmer test Ⅰ (S Ⅰ t),tear film break-up time (BUT),meibomian gland findings,the location of the keratopathy,the characteristics of keratophthy before and after fluorescein staining.The treating were given,including cessating of the original drugs,applying corneal repair promotion and anti-inflammatory drugs as well as the comprehensive treatment for meibomian gland embolization and dry eye,such as the hot packs and massage in the eyes with meibomian gland dysfunction and a tear dot embolization therapy in the eyes with S Ⅰ t ＜ 5 mm and BUT＜5 s.Paired t test and repeated measured one-way analysis of variance in SPSS 17.0 software were used to compare the BCVA,BUT and S Ⅰ t outcomes.The correlation between corneal repair duration and S Ⅰ t results was analyzed by Pearson linear correlation analysis.Results The primary cause of drug-induced keratopathy was irrational use of drugs,including antiviral drugs,antibiotics,hormone,antiallergic,lowering-intraocular pressure drugs,turn for 23 eyes,12 eyes,10 eyes,1 eye and 1 eye,respectively.Improper route of administration included 25 cases of overuse of eye drops and 6 cases of subconjunctival injection.BCVA was 0.69 ± 0.28 1 month after treatment,which was significantly improved in comparison with before treatment (0.32 ± 0.26) (t =11.02,P ＜ 0.01).Clinical manifestations included corneal epithelial diffusive and point-like roughness,corneal epithelial defect and even corneal ulcer for severe cases,corneal edema,Descemet membrane folds and partially visible corneal filiform.Drug-induced keratophthy was mainly located in the central and lower cornea.Comprehensive therapy was effective with the treating duration about 1 week to 8 weeks.A negative correlation was found between the corneal restore duration and S Ⅰ t results (r =-0.835,P＜0.01).Conclusions Corneal changes secondary to topical medications may affect all layers of the cornea.Clinicians should be mindful of drug-induced ocular disorders.The early diagnosis of druginduced keratopahthy depends on the medical history and clinical features.A comprehensive treating based on ocular surface conditions is available.

Objective To explore the expression and significance of let-7a2 in lung adenocarcinoma.Methods To adapt technology of realtime PCR with reference of U6, the expression of let-7a2 in tumor tissues and normal tissues was detected in 15 cases of resected lung adenocarcinoma. The laboratorial data were analysed by measure of 2~(-△△CT) method. The analysed data were managed by statistics. Results The expression of let-7a2 in lung adenocarcinoma tissue was significantly lower than that in lung normal tissue (0.8733±0.4821 vs 2.4527±1.0111). This difference possess markedness significance(t=6.816, P<0.001).Conclusion The expression of let-7a2 in lung adenocarcinoma is obviously lower and is concerned with the course of lung cancer evolution.

Objective To investigate the effects of contrast dose and region of interest (ROI) depth on quantitative analysis of liver by contrast-enhanced ultrasound (CEUS) during clinical application.Methods After bolus injection of contrast agent,the change of quantitative parameters [including echo mean(EM),rise time(RT),peak intensity(PI),mean transit time(MTT),area under the curve(AUC),time from peak to one half(TPH),wash in slope(WIS),time to peak(TTP)] of time-intensity curves were analyzed based on groups from different doses (1.0 ml and 1.6 ml) and different depth (＜30 mm,30-60 mm,and ≥60 mm).Results MTT and TPH were increased with dose increasing from 1.0 ml to 1.6 ml (P＜0.05).With the dose 1.0 ml,TPH,WIS,PI,AUC and MTT showed significant difference when the depth of the ROI changes (P ＜0.05),with the depth increased,TPH,WIS,PI,and AUC all decreases and MTT increases.For all the other parameters,no significant changes were found (P ＞0.05).Conclusions CEUS and its imaging process can directly influence the accuracy of the parameters from the quantitative analysis.Standardization of contrast agent with predefined dose and depth can potentially facilitate future clinical studies in liver CEUS.

Background Recent researches show that oxidative stress is involved in the progress of keratoconus.Nuclear factor-E2-related factor 2-antioxidant response element (Nrf2-ARE) pathway plays a critical role in the defense against oxidative stress,but its function in keratoconus is unclear.Objective To investigate the differences of Nrf2-ARE signaling activation and matrix degenerating enzymes between keratoconus and normal corneal stromal cells.Methods Corneal stromal cells were isolated from keratoconus and normal cornea by using dispase and collagenase digestion.The cells were treated with hydrogen peroxide (H2O2) to mimic in vivo oxidative stress condition.Reactive oxygen species (ROS) production was measured by fluorescence substrate DCHF-DA incubation.Nrf2 level and the expression of Nrf2-ARE downstream antioxidant genes were analyzed by Western blot and real-time quantitative-PCR(RT-qPCR).The activity of matrix degenerating enzymes,including urokinase-type plasminogen activator (uPA)-uPA receptor (uPAR) system and matrix metalloproteinase-2 (MMP-2) were assessed by Western blot and gelatin zymography respectively.Results In normal culture,keratoconus corneal stromal cells assumed increased basal ROS and Nrf2 level when compared with normal cells(t =18.155,P＜0.01).However,after H2O2 treatment,the keratoconus corneal stromal cells showed increased ROS production,while decreased Nrf2 translocation and no significant difference in expression levels of Nrf2-ARE downstream antioxidant genes (Nrf2:t =62.123,P＜ 0.01 ; (nicotinamide adenine dinucleotide phosphate quinine oxidoreductase-1 [NQO-1]:t =2.209,P =0.092 ; hemo oxygenase-1 [HO-1]:t =0.293,P =0.784 ; superoxide dismutase [SOD2]:t =0.749,P =0.495).The contents of uPA-uPAR and the activity of MMP-2 also showed a higher level in keratoconus corneal stromal cells than normal cells,with significant differences between them (t =19.164,15.458,4.818,all at P＜0.01).Conclusions The defect of Nrf2-ARE signaling activation exists in the keratoconus corneal stromal cells,and correlats with the abnormal expression level of stromal degeneration enzymes,which suggests that the defect of Nrf2-ARE signaling activation may be involved in the progression of keratoconus.

Objective To analyze the distribution, diversity, and the sensitivity of common pathogen species to antifungal drugs in fungal keratitis during 2000 - 2006 in Shandong province. Methods Samples from corneal serapings and corneal buttons from keratoplasties were used for fungal culture and identification. The distribution and diversity of the pathogens in different years were analyzed. The sensitivity of some common species to antifungal drugs were tested. Results A total of 898 patients were diagnosed as fungal keratitis, in which 770 (85.7%) were positive in fungal culture. Pathogens in 748 cases caused by fungal infection were identified, of which 547 cases were caused by Fusarium (71.0%). The prevelence of Fusarium from 2000 to 2006 was 75.5%, 72.8%, 71.5%, 75.2%, 76%, 68. 8% and 56.4% respectively. Eighty-four cases were caused by Aspergillus( 10.9% ) , which accounted for 15.1% , 15.2% , 13.1%, 10. 2%, 10. 4% , 8.0% and 6. 9% of the cases from 2000 to 2006, respectively. Seventy-four eases were caused by Alternaria (9. 6% ), the percentage of which during 2000 -2006 was 1. 9%, 3.3%, 3.7%, 6. 6%, 8. 8%, 12. 0 and 29. 7% respectively. In genus Fusarium, 10 species were found and the most common species were Fusarium Solani ( 33. 8% ), Fusarium oxysporum ( 28.2% ) and Fusarium moniliforme( 27.4% ). Six species were identified in genus Aspergillus, with Aspergillus flavus (53.6%) and Aspergillus fumigatus (39.3% ) being the most common species. The geometric mean MICs of amphotericin B, terbinafin, itraconazole, 5-flucytosine and fluconazole against the common species were 0. 647, 0. 714, 1. 624, 15. 108 and 27. 070 μg/ml, respectively. The mean MICs of the above five antifungal drugs against Fusarium were 0. 899, 0. 893, 3.077, 37. 124 and 21. 687 μg/ml, respectively, and for Aspergillus, the mean MICs of these agents were 0.794, 0.591, 0.416, 23.973 and 7.127 μg/ml, respectively; for Alternaria,the MICs were 0. 409,0. 479,0. 433,11. 655 and 7. 104 μg/ml ,respectively. Conclusions Genus Fusarium, Aspergillus and Alternaria were the predominant pathogens in fungal keratitis in Shandong Province. During 2000 -2006, Fusarium was the most common one, followed by Aspergillus. The percentage of Aspergillus decreased annually. The third most common pathogen, Alternaria increased during the period. The geometric mean MICs of amphotericin B and terbinafin against Fusarium were lower than those of itraconazole, 5-flucytosine and fluconazole. The MICs of these five anti-fungal drugs against Aspergillus and Alternatia were lower than those for Fusarium.

Objective:To investigation of the long-term-siRNA treatment with HBV transgene mice on inhibit replication of hepatitis B virus .Methods:The constructed siRNA expressed vectors was transfected HBV transgene mice by hydrodynamics -based in-jection via vena caudalis .Different groups were set including:specificity siRNA groups ( pSilencer5.1/C2,pSilencer4.1/C2,pSilenc-er3.1/C2),PBS group and negative vector group (n=10).The effect was observed in different periods (6 d,21 d,1 months,3 months, 6 months and 9 months after injection ) .HBsAg was analyzed by Chemiluminescence method , HBV-DNA was analyzed by real time quantitative PCR ( RQ-PCR) .Results:Compared with the PBS group , specificity siRNA groups showed decreased levels of HBsAg and HBV-DNA (P<0.05).Negative vector group did not show such changes ,there were no significant differences (P>0.05).Conclu-sion:The siRNA based on the expression vector can suppress the expression and replication of HBV in HBV transgene mice .The inhi-bition effects of long-term-siRNA treatment was specific .

Objective:To assess the changes in corneal endothelial cell density (CD) and morphology in patients with different stages of keratoconus.Methods:A cross-sectional study was conducted.One hundred and nineteen patients (199 eyes) with keratoconus who were treated in the Eye Hospital of Shandong First Medical University were included from March 2018 to October 2021.The 199 eyes were classified into stage Ⅰ (111 eyes of 58 cases), stage Ⅱ (41 eyes of 30 cases), stage Ⅲ (47 eyes of 31 cases) keratoconus groups according to the Amsler-Krumeich classification.In the same period, 25 age- and sex-matched healthy subjects (50 eyes) were enrolled as a normal control group.Corneal topography and anterior segment parameters such as keratometry (K), central corneal thickness (CCT), thinnest corneal thickness (TCT), anterior chamber depth (ACD), corneal diameter and corneal volume were obtained by Pentacam 3-dimensional anterior segment imaging and analysis system.The corneal endothelial CD, percentage of hexagonal cells (6A), average cell area (AVE), maximum cell area (MAX), minimum cell area (MIN), cell area standard deviation (SD) and cell area coefficient of variation (CV) in the central area were evaluated by non-contact specular microscopy.The correlation between corneal endothelial CD, morphological parameters and corneal topographic parameters was analyzed by Spearman rank correlation.This study adhered to the Declaration of Helsinki and was approved by the Ethics Committee of Shandong Eye Hospital (No.SDSYKYY201803). All patients were informed of the purpose and methods of the study and written informed consent was obtained before any medical examination.Results:The CD of the normal control group and stage Ⅰ, Ⅱ, Ⅲ keratoconus groups was 2 941(2 809, 3 072), 2 825(2 667, 3 030), 2 747(2 475, 2 903) and 2 370(2 142, 2 525) cells/mm 2, respectively.With the progression of keratoconus, CD decreased gradually, and there was a significant difference in CD among the four groups ( H=94.862, P<0.001). There were significant differences in CV and 6A among the four groups ( H=45.018, 20.421; both at P<0.001). CV was significantly higher in stage Ⅲ keratoconus group than that of the normal control group and stage Ⅰ and Ⅱ keratoconus groups and 6A was significantly lower in stage Ⅲ keratoconus group than that of the normal control group and stage Ⅰ keratoconus group (all at P<0.05). With the progression of keratoconus, MAX, MIN, AVE and SD increased gradually, and there were significant differences in MAX, MIN, AVE and SD among the four groups ( H=37.905, 32.437, 110.182, 72.941; all at P<0.001). MAX and MIN in stage Ⅲ keratoconus group were significantly higher than those in stage Ⅰ keratoconus groups and normal control group (all at P<0.05). AVE and SD in stage Ⅲ keratoconus group were significantly higher than those in normal control group and stage Ⅰ and Ⅱ keratoconus groups (all at P<0.05). In patients with keratoconus, CD was moderately positively correlated with CCT ( rs=0.47, P<0.001) and TCT ( rs=0.53, P<0.001), and was moderately negatively correlated with mean keratometry (Km) ( rs=-0.59, P<0.001).6A was weakly positively correlated with CCT ( rs=0.18, P=0.01) and TCT ( rs=0.22, P=0.002), and was weakly negatively correlated with Km ( rs=-0.32, P<0.001). CV was weakly negatively correlated with CCT ( rs=-0.35, P<0.001) and TCT ( rs=-0.37, P<0.001), and was moderately positively correlated with Km ( rs=0.48, P<0.001). There was no correlation between CD, CV, 6A and ACD, or corneal volume. Conclusions:As the keratoconus progresses, the cornea protrudes and becomes thinner with CD and 6A decreasing while CV increasing.Corneal topographic parameters are related to the density and morphology of corneal endothelial cells.

OBJECTIVETo test the immunosuppressive effect of cyclosporine (Cs) in a polymer placed in the anterior chamber of corneal allograft recipients.

METHODSWistar inbred rats with vascularized corneas were recipients of corneal allografts from Sprague-Dawley donor rats. Rats underwent penetrating keratoplasty and were divided randomly into four groups: untreated control animals (UCA); Cs-polymer anterior chamber recipients (CPA); co-polymer subconjunctival recipients (CPS); and Cs-olive oil drop recipients (COO). Grafts were examined by slit lamp every 3 days and clinical conditions were scored. Cs concentration in the aqueous humor was assayed at 1, 2, and 4 weeks. At 1, 2 and 4 weeks after transplantation, the operated eyes were collected for histopathological evaluation of the grafts.

RESULTSThe median survival time of the allografts was 8.2 +/- 1.48 days for the UCA group, 11.4 +/- 2.50 days for the CPS group, and 17.0 +/- 2.00 days for the CPA group. There was a statistically significant difference (P < 0.05) between survival time of the allografts in the animals of the CPA group compared to the other groups of graft recipients. Significantly higher concentrations of Cs were found in the eyes given an anterior chamber implant of Cs-polymer, compared to other treatment groups or untreated rats. A transient inflammatory response in the anterior chamber was observed in the CPA group.

CONCLUSIONSCs-polymer placed in the anterior chamber significantly prolongs corneal allograft survival time in a high risk corneal graft rejection model. This intraocular delivery system may be a valuable adjunct for the suppression of immune graft rejection.

Animals ; Aqueous Humor ; metabolism ; Corneal Transplantation ; Cyclosporine ; administration & dosage ; metabolism ; Drug Delivery Systems ; Graft Survival ; Immunosuppressive Agents ; administration & dosage ; Male ; Rats ; Rats, Wistar ; Transplantation, Homologous

Objective:To investigate the risk factors and treatment outcome of recurrent Acanthamoeba keratitis (AK) after corneal transplantation. Methods:A serial case-observational study was carried out.Twenty-eight eyes of 28 patients with AK who underwent corneal transplantation in Shandong Eye Hospital from January 2012 to January 2019 were enrolled.All the eyes received corneal transplantation from failing to respond to topical and systemic anti- Acanthamoeba medical therapy, including 13 eyes that received penetrating keratoplasty (PKP) and 15 eyes that received lamellar keratoplasty (LKP). The corneal lesion was removed by a trephine with a diameter of 0.5 mm over infiltration area during PKP or LKP.The clinical features of recurrent AK were summarized, including recurrence time, site and signs, and the risk factors of AK recurrence were analyzed.Local and systemic anti- Acanthamoeba medical therapy was performed in all relapsed eyes, and secondary surgery was performed for the eyes with poor response to medication.The therapeutic outcome of recurrent AK was evaluated.The study adhered to the Declaration of Helsinki.This study protocol was approved by an Ethics Committee of Shandong Eye Hospital (No.201112). Results:In the 28 eyes, 7 eyes (25%) appeared recurrent AK after keratoplasty, including 2 eyes after PKP and 5 eyes after LKP.There was no significant difference in the recurrence rate between the two methods ( P=0.396). The recurrence rate of eyes that had used glucocorticoids drugs before operation was 57.14% (4/7), which was significantly higher in comparison with 14.29% (3/21) of eyes without glucocorticoids before surgery ( P=0.043). The recurrence rate of eyes with ulcer diameter ≥8.2 mm was 50.00% (5/10), which was significantly higher than 11.11% (2/18) of eyes with ulcer <8.2 mm ( P=0.036). The recurrent lesions began at the edge of implant bed accounted for 85.71% (6/7), and the recurrent lesions located below graft accounted for 14.29% (1/7). In 7 eyes with recurrent AK, 6 eyes were completely cured.Among recurrent AK eyes after LKP, 2 eyes were cured by long-term medical therapy, and 2 eyes were cured by extended-diameter LKP, and another 1 eye was cured by conjunctival flap covering surgery.One eye with recurrent AK after PKP was cured by extended-diameter PKP. Conclusions:The risk factors of recurrent AK after surgery are application of glucocorticoids before surgery and big lesions.Recurrent AK after surgery is curable by individualized therapy targeting to different clinical characteristics.