Structural measurements and vessel density of spectraldomain optic coherence tomography in early,moderate,and severe primary angle-closure glaucoma

AIM:To compare the macular ganglion cell-inner plexiform layer(GCIPL)thickness,retinal nerve fiber layer(RNFL)thickness,optic nerve head(ONH)parameters,and retinal vessel density(VD)measured by spectral-domain optical coherence tomography(SD-OCT)and analyze the correlations between them in the early,moderate,severe primary angle-closure glaucoma(PACG)and normal eyes.METHODS:Totally 70 PACG eyes and 20 normal eyes were recruited for this retrospective analysis.PACG eyes were further separated into early,moderate,or severe PACG eyes using the Enhanced Glaucoma Staging System(GSS2).The GCIPL thickness,RNFL thickness,ONH parameters,and retinal VD were measured by SD-OCT,differences among the groups and correlations within the same group were calculated.RESULTS:The inferior and superotemporal sectors of the GCIPL thickness,rim area of ONH,average and inferior sector of the retinal VD were significantly reduced(all P<0.05)in the early PACG eyes compared to the normal and the optic disc area,cup to disc ratio(C/D),and cup volume were significantly higher(all P<0.05);but the RNFL was not significant changes in early and moderate PACG.In severe group,the GCIPL and RNFL thickness were obvious thinning with retinal VD were decreasing as well as C/D and cup volume increasing than other three groups(all P<0.01).In the early PACG subgroup,there were significant positive correlations between retinal VD and GCIPL thickness(except superonasal and inferonasal sectors,r=0.573 to 0.641,all P<0.05),superior sectors of RNFL thickness(r=0.055,P=0.049).More obvious significant positive correlations were existed in moderate PACG eyes between retinal VD and superior sectors of RNFL thickness(r=0.650,P=0.022),and temporal sectors of RNFL thickness(r=0.740,P=0.006).In the severe PACG eyes,neither GCIPL nor RNFL thickness was associated with retinal VD.CONCLUSION:The ONH damage and retinal VD loss appears earlier than RNFL thickness loss in PACG eyes.As the PACG disease progressed from the early to the moderate stage,the correlations between the retinal VD and RNFL thickness increases.

Inhibition of LOX-1 alleviates the proinflammatory effects of high-mobility group box 1 in Aspergillus fumigatus keratitis

AIM: To investigate the inflammatory amplification effect of high-mobility group box 1(HMGB1) in Aspergillus fumigatus(A. fumigatus) keratitis and the relationship between lectin-like oxidized low-density lipoprotein receptor 1(LOX-1) and HMGB1 in keratitis immune responses.METHODS: Phosphate buffer saline(PBS), and Boxb were injected into BALB/c mice subconjunctivally before the corneas were infected with A. fumigatus. RAW264.7 macrophages and neutrophils were pretreated with PBS and Boxb to determine the HMGB1 inflammatory amplification effects. Abdominal cavity extracted macrophages were pretreated with Boxb and Poly(I)(a LOX-1 inhibitor) before A. fumigatus hyphae stimulation to prove the the relationship between the two molecules. LOX-1, interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), macrophage inflammatory protein-2(MIP-2) and IL-10 were assessed by polymerase chain reaction and Western blot.RESULTS: Pretreatment with Boxb exacerbated corneal inflammation. In macrophages and neutrophils, A. fumigatus induced LOX-1, IL-1β, TNF-α and MIP-2 expression in Boxb group was higher than those in PBS group. Poly(I) treatments before infection alleviated the proinflammatory effects of Boxb in abdominal cavity extracted macrophages. Pretreatment with Boxb did not influence Dectin-1 mRNA levels in macrophages and neutrophils.CONCLUSION: In fungal keratitis, HMGB1 is a proinflammatory factor in the first line of immune response. HMGB1 mainly stimulates neutrophils and macrophages to produce inflammatory cytokines and chemokines during the immune response. LOX-1 participates in HMGB1 induced inflammatory exacerbation in A. fumigatus keratitis.

Epidemiological characteristics and risk factors of diabetic retinopathy in type 2 diabetes mellitus in Shandong Peninsula of China

AIM: To determine the epidemiological characteristics and estimate the risk factors of diabetic retinopathy (DR) in patients with type 2 diabetes mellitus (T2DM) in Shandong Peninsula of China. METHODS: The cases of T2DM admitted to Affiliated Hospital of Medical College of Qingdao University, Shandong Province, China, from January 2006 to December 2010 were retrospectively reviewed. The epidemiological characteristics of DR were estimated. The cases were divided into two groups according to degrees of retinopathy: non-DR group and DR group. Logistic regression analysis was used to study the related risk factors of DR. RESULTS: The prevalence of DR in patients with T2DM was 25.08% (834/3326). There was significant difference between the average age for men (59.08 +/- 15.43 years) and for women (62.92 +/- 18.19 years, P=0.0021). The majority of DR occurred in women (female: male ratio=1.76:1, P<0.0001). The incidence rate of DR in urban (489/834) was higher than that in rural area (345/834, P<0.0001). In 834 DR patients, the mean duration of T2DM was 8.90 +/- 4.15 years (range: 0-16 years); 440 people (52.76%) had received varying degrees of health education about prevention and primary care of DM; and 473 people (56.71%) suffered from other DM complications confirmed at the same time. In addition, the incidence rate of monocular (551/3326) and binocular retinopathy (283/3326) were statistically different (P<0.0001). Factors associated (p<0.05) with the presence of DR included old age, lower health educational level, intraocular surgery history, longer duration of T2DM, accompanying with other DM complications, no standard treatment procedure, lower body mass index (BMI) and higher fasting plasma glucose (FPG), glycated hemoglobin A(1)C (HbA(1)C), urine albumin (UA), total cholesterol (TC), low-density-lipoprotein cholesterol (LDL-C). The risk factors (P<0.05) independently associated with the presence of DR were: longer duration of T2DM, lower health educational level, higher FPG, higher UA, lower BMI and higher IC. CONCLUSION: DR is highly prevalent in the patients with T2DM in Shandong Peninsula of China. Besides blood glucose, many factors are associated with the present and development of DR.

Form-deprivation myopia induces decreased expression of bone morphogenetic protein-2, 5 in guinea pig sclera

AIM: To identify the presence of various bone morphogenetic proteins(BMPs) and their receptors in normal sclera of human, rat and guinea pigs, and to determine whether their expression changed with form-deprivation myopia(FDM) in guinea pig sclera.METHODS: The expression of BMPs and BMP receptors were detected using reverse transcription polymerase chain reaction(RT-PCR) and immunofluorescence. Two-week-old guinea pigs were monocularly form-deprived with a translucent lens. After fourteen days induction of FDM, total RNA was isolated and subjected to RT-PCR to examine the changes of BMPs and BMP receptors in tissues from the posterior sclera. Western blotting analysis was used to investigate their changes in protein levels.RESULTS: Human sclera expressed m RNAs for BMP-2,-4,-5,-7,-RIA,-RIB and BMP-RII. Conversely, rat sclera only expressed m RNA for BMP-7 and BMP-RIB,while the expression of BMPs and BMP receptors in guinea pigs were similar to that of humans. Human sclera also expresses BMP-2,-4,-5,-7 in protein level.Fourteen days after the induction of myopia, significant decreased expressions for BMP-2 and BMP-5 in the posterior sclera of FDM-affected eyes(P 【0.05 vs internal control eyes).· CONCLUSION: Various BMPs were expressed in human and guinea pig sclera. In the posterior sclera,expressions of BMP-2 and BMP-5 significantly decreased in FDM eyes. This finding indicates that various BMPs as components of the scleral cytokines regulating tissue homeostasis and provide evidence that alterations in the expression of BMP-2 and BMP-5 are associated with sclera remodeling during myopia induction.

Natamycin in the treatment of fungal keratitis : a systematic review and Meta-analysis

AIM: To review published clinical studies examining the effect of natamycin in the treatment of fungal keratitis.METHODS: We selected the publications in CENTRAL,MEDLINE, EMBASE, CNKI, and CBM. This study systematically reviewed published randomized controlled trials(RCTs) that compared natamycin to other antifungal agents, and conducted feasible Meta-analysis of efficacy results using Revman 5.2 software.RESULTS: We included seven trials which were mainly carried out in developing countries of Asia, with five trials conducted in India, one each in China and Bangladesh. A total of 804 participants were randomized to following comparisons: 2% econazole versus 5%natamycin showed little difference in the effects of treatment of fungal keratitis [RR =0.99, 95% confidence interval(CI), 0.8 to 1.21]; chlorhexidine gluconate versus5% natamycin indicated that the results on healing of the ulcer at 21 d was less conclusive(RR=0.77, 95% CI, 0.55 to 1.08; I2=0%); 1% voriconazole versus 5% natamycin suggested that natamycin treatment appeared to be significantly better outcomes than voriconazole(regression coefficient =-0.18 log MAR; 95% CI,-0.30 to-0.05; P =0.006), especially in Fusarium cases(regression coefficient=-0.41 log MAR; 95% CI,-0.61 to-0.20; P 【0.001);natamycin versus fluconazole showed a significant difference in cure rate(χ2=5.048, P 【0.05) and natamycin group was more effective than fluconazole in average period of therapy(t =7.94, P 【0.01).CONCLUSION: Natamycin was a preferable choice in the treatment of fungal keratitis, especially in the early period of Fusarium cases.

Regulation of LOX-1 on adhesion molecules and neutrophil infiltration in mouse Aspergillus fumigatus keratitis

AIM:To determine whether lectin-like ox-LDL receptor(LOX-1)regulates adhesion molecules expression and neutrophil infiltration in Aspergillus fumigatus(A.fumigatus)keratitis of C57 BL/6 mice.METHODS:C57 BL/6 mice were pretreated with a neutralizing antibody to LOX-1(5μg/5μL)or control nonspecific IgG(5μg/5μL),LOX-1 inhibitor Poly-I(2μg/5μL)or PBS by subconjunctival injection.Fungal keratitis(FK)mouse models of C57 BL/6 mice were established by scraping corneal central epithelium,smearing A.fumigatus on the corneal surface and covering the eye with contact lenses.The corneal response to infection was assessed via clinical score.The mRNA levels of the adhesion molecules intercellular cell adhesion molecule-1(ICAM-1),vascular cell adhesion molecule-1(VCAM-1),P-selectin and E-selectin were tested in control and infected corneas by reverse transcriptionpolymerase chain reaction(RT-PCR).The protein levels of ICAM-1 were evaluated by immunofluorescence(IF)and Western blot.Neutrophils were extracted from the abdominal cavity of C57 BL/6 mice followed by pretreatment using antibody to LOX-1(10μg/mL)or control nonspecific IgG(10μg/mL),the Poly-I(4μg/mL)or PBS.The cells were then stimulated with A.fumigatus and tested mRNA and protein levels of lymphocyte function-associated antigen-1(LFA-1)using RT-PCR and Western blot.IF and myeloperoxidase(MPO)assays were used to assess neutrophil infiltration in mice corneas.RESULTS:Pretreatment of LOX-1 antibody or the Poly-I reduced the degree of inflammation of cornea and decreased the clinical FK score compared with pretreatment of IgG or PBS(both P<0.01).And these pretreatment also displayed an obvious decline in the mRNA levels of ICAM-1,VCAM-1,P-selectin,E-selectin and LFA-1 expression compared with control groups(all P<0.01).Furthermore,pretreated with LOX-1 antibody or Poly-I,the protein levels of ICAM-1 and LFA-1 also decreased compared with control groups(all P<0.05).Neutrophil infiltration in the cornea was significantly reduced after pretreatment of LOX-1 antibody or Poly-I compared with control groups by IF and MPO assays(both P<0.01).CONCLUSION:Inhibition of LOX-1 can decrease the expression of adhesion molecules and reduce neutrophil infiltration in A.fumigatus infected corneas of C57 BL/6 mice.

Production of interleukin-1β related to mammalian target of rapamycin/Toll-like receptor 4 signaling pathway during Aspergillus fumigatus infection of the mouse cornea

AIM：To elucidate the effect of rapamycin on regulating the production of interleukin（IL）-1β in Aspergillus fumigatus（A.fumigatus）-induced keratitis and to verify whether the expression of IL-1β in A.fumigatus keratitis is associated with the mammalian target of rapamycin（mT OR）/Toll-like receptor 4（TLR4） signaling pathway.METHODS：Fungal keratitis mouse models of susceptible C57 BL/6 mice were established using A.fumigatus.The mice were subsequently treated with rapamycin.The protein levels of p-mT OR,TLR4,and IL-1β in normal and infected corneal tissue were measured by Western blot.The TLR4 and IL-1β m RNA levels were determined by real-time polymerase chain reaction（PCR）.RESULTS：In C57 BL/6 mice,rapamycin treatment decreased the clinical scores and production of the pro-inflammatory cytokine,IL-1β.The expression of TLR4,stimulated by A.fumigatus,was reduced as well when the mT OR signaling pathway was suppressed by rapamycin.CONCLUSION：Rapamycin is beneficial for the outcome of fungal keratitis and has an inhibitory effect expression of the inflammatory cytokine IL-1β.The inhibitory effect on IL-1β expression can be associated with the mT OR/TLR4 signaling pathway in A.fumigatus infection in mice.

Expression of dectin-1 during fungus infection in human corneal epithelial cells

AIM: To evaluate the expression of dendritic cell-associated C-type lectin-1(dectin-1) in human corneal epithelial(HCE) cells infected by fungus. · METHODS: A total of 20 cases of healthy donor corneas were group A,and 20 patients(20 eyes) suffered from fungal keratitis(FK) composed group B. Real-time qPCR and immunohistochemistry were applied to detect dectin-1 expression in corneal epithelium of both groups. HCE cells were cultured with aspergillus fumigatus(AF) antigens in vitro. The expression of dectin-1 mRNA was measured by real-time qPCR at the stimulation of 0,4,8 and 24h separately. Dectin-1 protein was detected by immunocytochemistry at 0 and 24h separately. ·RESULTS: Dectin-1 expressed in corneal epithelium of normal persons and FK patients. Vitro cellular experiment showed that the expression of dectin-1 mRNA in HCE cells began to increase after stimulation of AF antigens at 4h,and dectin-1 protein expression increased after stimulation at 24h. · CONCLUSION: Dectin-1 expressed in corneal epithelium of normal persons. AF antigens stimulation can elevate the expression of dectin-1 in HCE cells in vitro.

Effect of corneal graft diameter on therapeutic penetrating keratoplasty for fungal keratitis

AIM: To evaluate the effect of corneal graft diameter on therapeutic penetrating keratoplasty(PKP) for fungal keratitis. METHODS: A total of 116 patients (116 eyes) suffered from fungal keratitis underwent PKP at the Affiliated Hospital of Medical College Qingdao University from May 2006 to May 2010. They were divided into two groups according to the corneal graft diameter. 64 eyes' corneal graft diameter was 8.00mm or larger and 52 eyes' graft diameter was smaller than 8.00mm. The follow-up time was 2 years. The postoperative visual acuity and complications were documented and compared. RESULTS: Sixty-two (96.88%) eyes and fifty (96.15%) eyes preserved eyeballs respectively in two groups. There was no statistical difference in postoperative visual acuity (P = 0.961), corneal graft dear rate (P=0.132) or the incidence of recurred fungal infection (P=0.770) between two groups. But there was a higher incidence of graft rejection (P=0.020) and secondary glaucoma (P=0.039) in group with corneal graft diameter 8.00mm or larger. CONCLUSION: PKP is an effective treatment approach for fungal keratitis. There is a higher incidence of complications in large-diameter PKP for fungal keratitis.Effective, preventive and therapeutic measures can improve the prognosis.

Long-term postoperative outcomes of bilateral lateral rectus recession vs unilateral recession-resection for intermittent exotropia

AIM: To discuss the long-term postoperative results of bilateral lateral rectus recession(BLR) and unilateral lateral rectus recession-medial rectus resection(RR) in therapy of intermittent exotropia.METHODS: We retrospectively analyzed 213 cases of intermittent exotropia who underwent surgery between2008 and 2010. The patients were grouped into BLR group and RR group. Motor outcomes were divided into three groups on the basis of the angle of deviation after surgery: overcorrection(esotropia/phoria 】5△), orthophoria(esotropia/phoria ≤5△to exotropia/phoria ≤10△), and undercorrection/recurrence(exotropia/phoria 】10△).Titmus test was used to evaluate stereoacuity, the stereoacuity 【800s of arc meaned the patients had stereopsis. Surgical outcome including motor criteria and sensory status were compared at postoperative 6, 12,24 mo and at 36 mo examination between groups. RESULTS: At 12, 24 mo after surgery, the motor outcomes had no difference(P 】0.05) between groups.However, the motor outcomes at 6, 36 mo were signally different in each group, indicating the success rate in RR group at 6mo was higher than that in BLR group(83.02%vs 82.24%, P 【0.05) but the result was contrary at the 3y examination(60.75% vs 43.40%, P 【0.05). No statistical significance were found in the sensory outcomes between the groups at mean of 3.7y follow-up.CONCLUSION: The motor outcomes in RR group were better than in BLR group at 6mo after surgery, while the3 y outcomes were better in BLR group. This may be due to the recurrence rate of the BLR was lower than the RR group’s.

Role of the IL-33/ST2/p38 signaling pathway in the immune response of corneal epithelial cells to Aspergillus fumigatus infection

AIM: To investigate the expression of interleukin(IL)-33 in the cornea and human corneal epithelial cells(HCECs) exposed to Aspergillus fumigatus(A. fumigatus), and to determine the function of IL-33/ST2/p38 signaling pathway in the immune response of corneal epithelial cells to A. fumigatus infection.METHODS: The mRNA and protein expression of IL-33 in HCECs and mice corneas were examined by quantitative real-time reverse transcription polymerase chain reaction(q RT-PCR) and Western blot analysis, respectively. IL-33 expression was also detected in cornea samples from healthy donors and patients with fungal keratitis with immunohistochemistry. The cultured HCECs were treated with inactive A. fumigatus hyphae at various concentrations with or without recombinant human IL-33 protein, soluble recombinant ST2 protein, specific ST2 neutralizing antibody, or the mitogen-activated protein kinase(MAPK) p38 inhibitor SB203580 for evaluation of the expression and activation of IL-33/ST2/p38 signaling in the regulation of proinflammatory cytokines. The production levels of IL-6 and IL-1β were determined by qR T-PCR and enzymelinked immunosorbent assay(ELISA). The proliferation of HCECs was determined by a Cell Counting Kit-8(CCK8) assay and cell count.RESULTS: IL-33 expression levels increased in the corneal tissues of patients with fungal keratitis and in mice corneas of experimental A. fumigatus infection,as well as in HCECs with infection of A. fumigatus. A. fumigatus strongly stimulated HCECs-generated proinflammatory cytokine(IL-6 and IL-1β) production at both the mRNA and protein levels. This production of proinflammatory mediators stimulated by A. fumigatus was further stimulated by IL-33 and was prevented by soluble ST2 protein or ST2 neutralizing antibody. Moreover, IL-33 naturally promoted the p38 phosphorylation induced by A. fumigatus, which was suppressed by soluble ST2 protein. The MAPK p38 inhibitor SB203580 also inhibited the A. fumigatus-induced proinflammatory cytokine production. IL-33 administration for 48 h and 72 h promoted the proliferation of HCECs, which was attenuated by treatment with soluble recombinant human ST2 protein.CONCLUSION: A. fumigatus elevates IL-33 expression in human and mice corneas and HCECs. Thus, IL-33/ST2/p38 signaling may play an important role in amplifying the immune response of corneal epithelial cells to A. fumigatus infection. Besides, IL-33 promotes the cell proliferation of HCECs via its receptor ST2. These findings suggest a novel autocrine mechanism of amplification of the fungalinduced inflammatory response in the corneal epithelium, highlighting a potential therapeutic target for fungal keratitis.

Fungus induces the release of IL- 8 in human corneal epithelial cells, via Dectin-1-mediated protein kinase C pathways

AIM: To identify whether Aspergillus fumigatus(A.fumigatus) hyphae antigens induced the release of interleukin-8(IL-8) in anti-fungal innate immunity of cultured human corneal epithelial cells(HCECs) and determine the involvement of intracellular signalling pathways. METHODS: HCECs were treated with A. fumigatus hyphae antigens with different concentrations and time.The cytoplasmic calcium of HCECs were assessed by fluorescence imaging. Western blot was used to detect the expression of Ca2 +-dependent protein kinase C(PKC). The IL-8 levels were determined by specific human IL-8 enzyme-linked immunosorbent assay(ELISA) and reverse transcriptase polymerase chain reaction(RT-PCR). Using a series of pharmacological inhibitors, we examined the upstream signalling pathway responsible for IL-8 expression in response to A.fumigatus hyphae antigens. RESULTS: Cells exposed to A. fumigatus hyphae antigens showed higher level of IL-8 m RNA expression and protein production. We demonstrated here that stimulation of HCECs with A. fumigatus hyphae triggers an intracellular Ca2 +flux and results in the activation of Ca2 +-dependent PKC(α, βⅠ and βⅡ) which can be attenuated by pre-treatment of cells with laminarin,suggesting that Dectin-1 signals pathway induced cytoplasmic calcium and influence the activation of PKC in HCECs. Inhibitors of Ca2 +-dependent PKC(Ro-31-8220 and Go6976) significantly abolished hyphae-induced expression of IL-8.CONCLUSION: Our findings suggest that A. fumigatushyphae-induced IL-8 expression was regulated by the activation of Dectin-1-mediated Ca2 +-dependent PKC in HCECs.

Expression of S100B during the innate immune of corneal epithelium against fungi invasion

AIM： To explore the expression of SIOOB in corneal epithelial cells under ,Aspergillus stimulation both in vivo and in vitro. METHODS： Immortalized human corneal epithelial cells （HCECs） were exposed to inactive #lsperg///us fumigatus （A. fumigatus） conidia at 0, 4, 8, 12, 16, and 24h respectively. The corneas of Wistar rats were exposed to active A. fumigatus at 0, 12, 24, 48h and the normal rat corneas were used for normal control. The mRNA level of S100B was evaluated by real time quantitative reverse transcription-polymerase chain reaction （qRT-PCR）. Sl00B protein expression in cornea epithelium was detected by immunohistochemical/immunocytochemical staining （IHC/ICC）. RESULTS： Histopathology revealed a significant inflammatory cell infiltration in fungal keratitis human and rat cornea. Corneal epithelial cells didn＇t express or rarely express S100B at baseline. A. fumigatus significantly induced S100B mRNA expression in cultured corneal epithelial cells in a time depended manner in vitro the mRNA began to rise significantly at 8h in vitro （P〈0.05） and continue to rise as time prolonged （P〈0.01）. in vivo S100B mRNA level was low in the normal corneas. However, it was increased in keratitis corneas from 12h after infection （P〈0.05） and reached to a peak at 24h （P〈0.001）. Immunochemistry revealed an obvious staining in fungal keratitis corneas as well as immortalized HCECs compared to the normal ones respectively, indicating an increased expression of SlOOB protein. CONCLUSION： S100B exists in corneal epithelial cells and is over-expressed under A. fumigatus stimulation. Sl00B may play an important role in the innate immune response of the corneal epithelium during A. fumigatus infection.

The role of LOX-1 on innate immunity against Aspergillus keratitis in mice

AIM：To explore the effects of lectin-like ox-LDL receptor（LOX-1） on innate immunity against Aspergillus fumigatus（A.fumigatus） in mice cornea.METHODS：The mRNA levels of LOX-1 were tested in normal and A.fumigatus infected corneas of C57BL/6and BALB/c mice.The expression of LOX-1,pro-inflammatory cytokines TNF-α,CXCL1 and IL-6,antiinflammatory cytokines IL-10,and matrix metalloproteinase 9（MMP9） were tested with treatment with LOX-1 neutralizing antibody or control IgG in A.fum/gatus infected corneas of C57BL/6.Macrophages and neutrophils were extracted from susceptible C57BL/6mice,and pretreated with LOX-1 neutralizing antibody or IgG,then stimulated with A.fum/gatus.The mRNA levels of LOX-1,TNF-α,CXCL1,IL-6,IL-10 and MMP9 were evaluated by polymerase chain reaction.RESULTS：The expression of LOX-1 was significantly increased in C57BL/6 mice corneas after A.fum/gatus infection compared with BABL/c mice.After treatment with LOX-1 neutralizing antibody,the expression of LOX-1,TNF-α,CXCL1,IL-6,MMP9 and IL-10 in C57BL/6corneas were significantly decreased compared with treatment with control IgG;the expression of LOX-1,CXCL1,IL-6 and IL-10 were significantly decreased in macrophages,while TNF-α and MMP9 expressions had no change;LOX-1,TNF-α,CXCL1,IL-6,MMP9 and IL-10 expressions were significantly decreased in neutrophils.CONCLUSION：The expression of LOX-1 can affect the expression of pro-inflammatory and anti-inflammatory cytokines in fungal infected corneas,macrophages and neutrophils of C57BL/6.LOX-1 inhibition rebalances the inflammatory response of fungal keratitis in mice.

Anti-inflammatory effects of astaxanthin against fungal keratitis

AIM:To characterize effect of astaxanthin(ASX)in Aspergillus fumigatus(A.fumigatus)induced keratitis in mouse model.METHODS:In vivo,fungal keratitis mouse model was established in C57BL/6 mice using A.fumigatus,followed by ASX or dimethyl sulfoxide(DMSO)treatment.Clinical responses were evaluated by clinical score and myeloperoxidase(MPO)assay.Inflammatory cytokines were assessed by reverse-transcription polymerase chain reaction(RT-PCR),Western blot,immunofluorescence,and enzyme-linked immuno sorbent assay(ELISA).RESULTS:In animal model,ASX improved corneal transparency and clinical response,suppressed the expression of inflammatory cytokine like IL-1β,TNF-α,and HMGB-1.Neutrophil levels have been shown to decrease in ASX-treated cornea by immunofluorescence and MPO.TLR2 and TLR4 levels were lower in ASX-treated group than DMSO-treated.CONCLUSION:ASX can suppress inflammatory response and reduce inflammatory cytokine production in mice model with A.fumigatus keratitis.

Early expression of surfactant proteins D in Fusarium solani infected rat cornea

AIM: To investigate the early expression of surfactant proteins D(SP-D) in Fusarium solani infected rat cornea. METHODS: Wistar rats were divided into group A, B and C randomly. The right eyes were chosen as the experiment one. Group A was control group. Group B was not inoculated with Fusarium solani. Group C was taken as fusarium solani keratitis model. Five rats in group B and C were executed randomly at 6, 12, 24, 48 and 96 hours respectively after the experimental model being established. The expression of SP-D was assessed through immunohistochemistry and reverse transcription polyrnerase chain reaction(RT-PCR). RESULTS: RT-PCR detected that the SP-D mRNA expression was low in the corneal of normal rats and group B. The expression of fungal infected cornea increased gradually and reached the peak at 24 hours in group C. The synchronous expression of group B and C were in significant difference (P<0.01). Immunohistochemisty discovered the protein of SP-D expression was increased gradually from 12 hours and reached the peak at 48 hours in group C. The synchronous expression of group B and C were also in significant difference (P<0.01). CONCLUSION: There exists SP-D in rat corneal tissue and the expression is significantly increased at the early period of fusarium solani infected cornea. SP-D may play a role in the early innate immunity response of the corneal resistance to Fusarium solani infection.

Characteristics of ocular abnormalities in gout patients

·AIM: To characterize the clinical features of ocular surface in gout patients in coastal area of Shandong Province in China. ·METHODS: A total of 380 consecutive gout patients were examined from January 2011 to May 2011. According to the course of gout, patients were divided into group A (【5 years), B (5 -10 years) and C (】10 years). Group D (control group) was consist of 50 healthy subjects. Eyelids, lateral canthus, medial canthus, palpebral conjunctiva, sclera and cornea, anterior chamber, lens, anterior vitreous were examined by slit lamp to find whether there were deposition of uric acid crystals, ocular vascular tortuosity, redness and subconjunctival hemorrhage. The ophthalmic exams of visual acuity, intraocular pressure, fundus were used to assess any gout-related eye disease. ·RESULTS: Uric acid crystals were found in 3 patients and the positions of the deposite were in corneal stroma, corneal epithelium and superficial stroma, and sclera respectively. The incidence was 0.79% . Dilatated and tortuous blood vessels in conjunctiva and sclera surface were found in 38 (23.8% ), 40 (44.0% ), 58 (45.0% ), 9 (18.0% ) patients in groups A, B, C and D, respectively. The differences between group B and D, group C and D were statistically significant( 【0.01, 【0.01).Transparent vesicles with metal -like reflected light in subconjunctiva were seen in 26 (16.2%), 29 (31.9%), 41 (31.8%), 2 (4.00%) patients in groups A, B, C and D, respectively. The differences between A and D, B and D, C and D were statistically significant ( 【0.05, 【0.01, 【0.01). Subconjunctival hemorrhage was found in all groups, the difference among the four groups showed no statistically significance. · CONCLUSION: Gout can cause ocular surface abnormalities, such as tophi deposition, subconjunctival transparent vesicles and hemorrhage, and vascular changes. These features have important clinical significance in early detection of the gout and prevention of eye injury. ·

Expression and role of aryl hydrocarbon receptor in Aspergillus fumigatus keratitis

●AIM:To observe the expression and role of aryl hydrocarbon receptor(Ah R)in the immune response of mouse cornea infected with Aspergillus fumigatus(A.fumigatus).●METHODS:Murine models of A.fumigatus keratitis were established by scraping the central epithelium of mouse cornea,daubing A.fumigatus on the cornea and covering with a contact lens.The mice were randomly divided into the control group and the A.fumigatus-infected(A.F.)group for 1,3 and 5 d respectively,which corneas were daily monitored by a slit lamp microscope and the clinical scores were also recorded timely after infection.In this study,immunofluorescence staining was used to detect the expression and localization of Ah R in mouse corneas,and the m RNA and protein of Ah R were detected by reverse transcription-polymerase chain reaction(RT-PCR)and Western blot.In addition,mouse peritoneal macrophages were stimulated by A.fumigatus with or without the pretreatment of Ah R antagonist CH223191 and Ah R agonist FICZ,and the tumor necrosis factor alpha(TNF-α),inducible nitric oxide synthase(i NOS),interleukin-10(IL-10)and Arg-1 m RNA were detected by RT-PCR.●RESULTS:According to the results of the slit light photography,it was clearly indicated that the corneal inflammation were the most severe and the clinical score became the highest as well on the 3 rd day after the infection of A.fumigatus.Contrasted with the control group,the expression of Ah R in the corneal epithelial cells infected with A.fumigatus was significantly increased detected by immunofluorescence staining.Ah R mainly expressed in the nucleus and cytoplasm of corneal epithelial cells.Consistent with the transcriptional level of Ah R m RNA,the expression level of Ah R protein reached the peak on the 3 rd day after infection which was detected by Western blot.Furthermore,RT-PCR showed that CH223191 up-regulated the expression of TNF-αand i NOS and down-regulated the expression of IL-10 and Arg-1 in peritoneal macrophages;inversely,FICZ reduced the expression of TNF-αand i NOS while elevated the expression of IL-10 and Arg-1.●CONCLUSION:Ah R is involved in the pathogenesis of A.fumigatus keratitis and induced immune protection in anti-A.fumigatus immune response by inhibiting M1 and increasing M2 phenotype macrophage-related inflammatory factors.

Dectin-1 expression at early period of Aspergillus fumigatus infection in rat's corneal epithelium

AIM:To investigate the expression of dendritic cell-associated C-type lectin-1 (dectin-1) at the early period of Aspergillus fumigatus infection in rat’s corneal epithelium. ·METHODS:A total of 72 Wistar rats were randomly divided into three groups:A, B and C. The right eyes were chosen as experimental eyes. Group A was control group. Rats in group B were not inoculated with Aspergillus fumigatus. Group C was taken as Aspergillus fumigatus keratitis model. Rats in group B and C (six from each group) were executed randomly at 4, 8, 16 and 24 hours after experimental model being established to assess the expression of dectin-1 mRNA through real-time PCR. Another six rats in group B and C were executed randomly at 24 hours to assess the expression of dectin-1 protein through immunohistochemistry. ·RESULTS:The results of real-time PCR indicated that dectin-1 mRNA expression was low in corneal epithelium of normal rats’. There was no significantly difference of dectin-1 mRNA expression in group A and B (P 】0.05). The expression of Aspergillus fumigatus infected corneal epithelium increased gradually after 8 hours in group C. The synchronous expression of group A and C had significant difference (P 【0.01). Immunohistochemisty discovered that dectin-1 receptor existed in normal rat’s corneal epithelium . Dectin-1 protein increased after 24 hours in group C. There was a significant difference of synchronous expression in group B and C(P【0.01). · CONCLUSION:Dectin-1 exists in rat’s cornealepithelium and its expression significantly increases at the early period of Aspergillus fumigatus infection. Dectin-1 is a pattern recognition receptor that expresses in corneal epithelium and involves in immune response to Aspergillus fungal keratitis.

Protein tyrosine phosphatase 1B regulates migration of ARPE-19 cells through EGFR/ERK signaling pathway

AIMTo evaluate whether protein tyrosine phosphatase 1B (PTP1B) contributed to initiate human retinal pigment epithelium cells (A)-19 migration and investigate the signaling pathways involved in this process.METHODSARPE-19 cells were cultured and treated with the siRNA-PTP1B. Expression of PTP1B was confirmed by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). AG1478 [a selective inhibitor of epidermal growth factor receptor (EGFR)] and PD98059 (a specific inhibitor of the activation of mitogen-activated protein kinase) were used to help to determine the PTP1B signaling mechanism. Western blot analysis verified expression of EGFR and extracellular signal-regulated kinase (ERK) in ARPE-19 cells. The effect of siRNA-PTP1B on cell differentiation was confirmed by immunostaining for &#x003b1;-smooth muscle actin (&#x003b1;-SMA) and qRT-PCR. Cell migration ability was analyzed by transwell chamber assay.RESULTSThe mRNA levels of PTP1B were reduced by siRNA-PTP1B as determined by qRT-PCR assay. SiRNA-PTP1B activated EGFR and ERK phosphorylation. &#x003b1;-SMA staining and qRT-PCR assay demonstrated that siRNA-PTP1B induced retinal pigment epithelium (RPE) cells to differentiate toward better contractility and motility. Transwell chamber assay proved that PTP1B inhibition improved migration activity of RPE cells. Treatment with AG1478 and PD98059 abolished siRNA-PTP1B-induced activation of EGFR and ERK, &#x003b1;-SMA expression and cell migration.CONCLUSIONPTP1B inhibition promoted myofibroblast differentiation and migration of ARPE-19 cells, and EGFR/ERK signaling pathway played important role in migration process.