The association of demodex infestation with pediatric chalazia.

PURPOSE: This study aimed to investigate the association of Demodex infestation with pediatric chalazia. METHODS: In a prospective study, 446 children with chalazia and 50 children with non-inflammatory eye disease (controls) who underwent surgical treatment were enrolled from December 2018 to December 2019. Patient ages ranged from 7 months to 13 years old. All patients underwent eyelash sampling for light microscope examination, and statistical correlation analysis between Demodex infestation and chalazia, including the occurrence, recurrence, and course of disease, morphological characteristics, and meibomian gland dysfunction (MGD) in chalazia patients was performed. RESULTS: Demodex was found in 236 (52.91%) patients with chalazia and zero control patients. Demodicosis was significantly more prevalent in chalazia patients than the control group (P < 1 × 10- 14). Recurrent chalazia (P = 0.006) and skin surface involvement (P = 0.029) were highly correlated with Demodex infestation. Demodicosis was also associated with multiple chalazia (P = .023) and MGD(P = .024). However, Demodex infestation was comparable in the course of disease (P = 0.15), seasonal change (P = 0.68) and blepharitis subgroups (P = 0.15). Within the group of chalazia patients who underwent surgical removal of cysts, 4 (0.9%) patients with concurrent demodicosis experienced recurrence. CONCLUSIONS: Demodex infestation was more prevalent in pediatric chalazia patients than healthy children, and was associated with recurrent and multiple chalazia. Demodicosis should be considered as a risk factor of chalazia. In children with chalazia, Demodex examination and comprehensive treatment of Demodex mites should be applied to potentially prevent recurrence.

Quantitative assessment and determinants of the papillary microvasculature in healthy subjects.

BACKGROUND: It is critical to monitor the optic disc's vessel density using Optical coherence tomography angiography (OCTA) and evaluate its determinants. In the current study, we investigate the superficial vessel density (VD) of the papillary microvasculature and its determinants in healthy subjects of Southern China. METHODS: This was a prospective, cross-sectional study. Superficial VD in healthy individuals' optic disc region was measured by OCTA. The factors associated with ocular and systemic parameters were analyzed using a generalized estimation equation (GEE) model. RESULTS: A total of 510 eyes of 260 healthy subjects were analyzed in the study. The total VD in the optic disc area was 17.21 ± 2.15 mm- 1 (95% CI, 17.02-17.40 mm- 1). The VD in the inner ring and the outer ring of the optic disc were significantly higher compared with the central ring, while the VD of the superior quadrant and inferior quadrant was significantly higher compared with the temporal and nasal quadrant. After adjusting for the ocular factors and systemic factors, AL (ß = - 0.4917, P = 0.0003), disc area (ß = - 0.3748, P = 0.0143), CMT (ß = - 0.0183, P = 0.0003) and SSI (ß = 1.0588, P < 0.001) were significantly associated with total VD of the optic disc. CONCLUSION: The mean total VD in the optic disc area was 17.21 ± 2.15 mm- 1 in healthy subjects, and the superior and inferior VD was significantly higher than the temporal and nasal VD. AL, disc area, CMT, and SSI may affect the total VD in the optic disc area and should be considered in clinical practice.

Efficacy of the WINROP algorithm for retinopathy of prematurity screening in Southern China.

AIM: To evaluate the predicting efficacy of severe retinopathy of prematurity (ROP) by the WINROP algorithm (http://winrop.com) in Southern China. METHODS: All preterm infants with the gestational age (GA) less than 32wk were included. Their ROP screening results and serial postnatal body weight were analysed retrospectively. Weekly body weight was entered into and measured by the WINROP system. The outcomes were analysed, and the sensitivity, specificity, positive predictive value and negative predictive value (NPV) were calculated. RESULTS: Totally 432 infants with a median GA of 30.0 (24.0-31.9)wk, and a median birth weight (BW) of 1360 (540-2700) g were included. Among these 432 infants, 50 were diagnosed as type 1 ROP but only 28 were identified by the WINROP algorithm. The sensitivity was 56% (28/50) and the NPV was 92% (252/274). However, for infants with BW <1000 g or GA <28wk, the sensitivity was 93.8% (15/16) and 93.3% (14/15), respectively. Meanwhile, with several postnatal complications added as additional risk factors, the sensitivity was increased to 96% (48/50). CONCLUSION: The sensitivity of the WINROP algorithm from the Southern Chinese cohort is not as high as that reported in developed countries. This algorithm is effective for detecting severe ROP from extremely small or preterm infants. Modification of the algorithm with additional risk factors could improve the predictive value for infants with a GA>28wk in China.

The role of soluble programmed death protein-1 (sPD-1) and soluble programmed death ligand-1 (sPD-L1) in rat corneal transplantation rejection.

BACKGROUND: Immunological rejection is one of the problems in corneal transplantation. Recently, some research found out that soluble programmed death protein-1 (sPD-1) and soluble programmed death ligand protein-1 (sPD-L1) play a significant role in immunologic suppression. OBJECTIVES: To explore expression of sPD-1 and sPD-L1 in a penetrative corneal transplantation model and its relationship with transplant rejection. MATERIAL AND METHODS: Autologous corneal transplantation rat models and allogeneic corneal transplantation rat models were used as the control group and the experimental group, respectively. Changes of the transplanted grafts were observed under a slit-lamp microscope. Hematoxylin-eosin (H&E) staining was applied to examine the histopathological features of the corneal grafts. Flow cytometry was used to analyze CD4+CD25+Treg in the serum and spleen. The sPD-1, sPD-L1, interleukin 10 (IL-10) and interleukin 4 (IL-4) levels in serum and the aqueous humor of the rats were detected using enzyme-linked immunosorbent assay (ELISA). RESULTS: After the operation, no transplant rejection occurred in the control group. Flow cytometry results showed that expressions of CD4+CD25+Treg in serum in the experimental group were lower than those in the control group (p < 0.05). The ELISA results showed that after the operation, sPD-1 and sPD-L1 expression levels in serum in the experimental group were higher than in the control group (all p < 0.05). After the operation, lL-10 and IL-4 content in serum in the experimental group was lower than in the control group (all p < 0.05). The sPD-1/sPD-L1 ratio in the experimental group was higher than in the control group. CONCLUSIONS: Increases of sPD-1 content and decreases of CD4+CD25+Treg, IL-10 and IL-4 levels may be involved in corneal allograft rejection. Dynamic detection of the content of sPD-1 and sPD-L1 in serum and aqueous humor after the operation would help in understanding the local immune response in a clinical setting and predicting the occurrence of corneal graft rejection.

The prognosis of trabeculectomy in primary angle-closure glaucoma patients.

AIM: To evaluate whether the level of thrombospondin-1 (TSP-1) in aqueous humor can predict the prognosis of trabeculectomy in patients with primary angle-closure glaucoma (PACG). METHODS: This case-control study involved 26 patients with PACG who experienced a failed trabeculectomy (case group) and 78 age- and sex-matched patients with PACG who underwent successful trabeculectomy (control group). Aqueous humor was collected at the time of trabeculectomy and tested for TSP-1 and TGF-ß2 levels with an enzyme-linked immunosorbent assay method. Logistic regression modeling was used to assess the risk factors for failed trabeculectomy. RESULTS: The mean TSP-1 aqueous concentrations were significantly higher in the case group (20.67±9.79 ng/mL) than the control group (5.17±2.29 ng/mL) (P<0.001). The transforming growth factor-ß2 (TGF-ß2) aqueous concentrations were significantly different between the case and control group, at 3633.25 and 1090.24 pg/mL, respectively (P<0.001). Logistic regression analysis revealed TSP-1 level as an independent risk factor for a failed trabeculectomy (OR=3.540; 95%CI=1.092-11.482). CONCLUSION: The aqueous humor TSP-1 and TGF-ß2 levels are higher in PACG eyes with failed trabeculectomy than with successful trabeculectomy at one year. The aqueous humor TSP-1 level is an independent risk factor associated with failed trabeculectomy.

Low concentration of sodium hyaluronate temporarily elevates the tear film lipid layer thickness in dry eye patients with lipid deficiency.

AIM: To investigate the effects of different concentrations of artificial tears on lipid layer thickness (LLT) and blink rate (BR) in dry eye patients. METHODS: This study included 106 eyes of 58 patients with dry eye. The lipid deficiency type was defined as the LLT baseline <75 nm. The LLT and BR were measured at baseline and 1, 5 and 15min after the instillation of 0.1% or 0.3% sodium hyaluronate (SH) eye drops by using the LipiView ocular surface interferometer. RESULTS: In the lipid deficiency group, the LLT increased from baseline at 1min post instillation. The LLT after the instillation of 0.1% SH was significantly higher than that after the instillation of 0.3% SH (P<0.001). The LLT returned to baseline at 15min post instillation of 0.1% SH and at 5min post instillation of 0.3% SH. In the non-lipid deficiency group, the LLT decreased from baseline at 1min and returned to baseline at 5min for both treatments. The BRs were not significantly different at different time points for both treatments. CONCLUSION: SH eye drops induce a short-term increase in LLT of patients with lipid deficiency. A low concentration of artificial tears have a stronger effect than a high concentration of artificial tears on the increase in LLT. In comparison, SH eye drops induce a transient and slight decrease in LLT of patients without lipid deficiency. A low concentration of artificial tears might be better for patients with lipid deficiency.

Choroidal thickness in pregnant women: a cross-sectional study.

AIM: To investigate choroidal thickness in pregnant women and compare the measurements with those of normal nonpregnant women. METHODS: Using enhanced depth imaging optical coherence tomography (EDI-OCT), choroidal thickness was measured at the fovea and at 1 mm and 3 mm superior, inferior, temporal, and nasal to the fovea in both healthy pregnant women and nonpregnant women. Pearson correlation analysis was performed to evaluate the relationships between subfoveal choroidal thickness (SFCT) and the demographic and ocular parameters. Pooled odds ratio (OR) and 95% confidence interval (CI) were calculated using fixed-effects model when Meta-analyses were conducted. RESULTS: Comparison of choroidal thickness between the groups showed that it was significantly greater in healthy pregnant women's eyes than in normal nonpregnant women's eyes at all locations except at 3 mm superior and 3 mm temporal from the fovea (P<0.05). The mean SFCT was 344.13±50.94 µm in healthy pregnant women's eyes and 315.03±60.57 µm in normal nonpregnant women's eyes, with a statistically significant difference (P=0.008). Pearson correlation analysis showed that age and axial length were significantly related to SFCT in healthy pregnant women, normal nonpregnant women, and all subjects. The results of our cross-sectional study were consistent with the results of the further Meta-analysis, with a pooled weighted mean difference (WMD) of 33.66 µm (95% CI: 26.16 to 41.15) for SFCT. CONCLUSION: Our results, along with the comprehensive Meta-analysis, suggest that choroidal thickness in healthy pregnant women is greater than that in normal nonpregnant women.

MiR-769 promoted cell proliferation in human melanoma by suppressing GSK3B expression.

MicroRNAs (miRNAs) are short, non-coding RNAs with post-transcriptional regulatory function, playing crucial roles in cancer development and progression of human melanoma. Previous studies have indicated that miR-769 was implicated in diverse biological processes. However, the underlying mechanism of miR-769 in human melanoma has not been intensively investigated. In this present study, we aimed to investigate the role of miR-769 and its target genes in human melanoma. We found that miR-769 expression was strongly increased in human melanoma cells and clinical tissues compared with their corresponding controls. Overexpression of miR-769 promoted cell proliferation in human melanoma cell line A375, whereas miR-769-in reverses the function. Glycogen synthase kinase-3 Beta (GSK3B), a potential target gene of miR-769, and was validated by luciferase assay. Further studies revealed that miR-769 regulated cell proliferation of human melanoma by directly suppressing GSK3B expression and the knockdown of GSK3B expression reversed the effect of miR-769-in on human melanoma cell proliferation. In summary, our data demonstrated that miR-769 might act as a tumor promoter by targeting GSK3B during development of human melanoma.

Simultaneous treatment of pterygium complicated with conjunctivochalasis: analysis of pterygium excision and conjunctival autotransplantation combined with sclera fixation.

BACKGROUND: This prospective study investigated the safety and efficacy of a therapeutic method of treating pterygium complicated with conjunctivochalasis, using pterygium excision and conjunctival autotransplantation combined with sclera fixation, followed by therapeutic contact lens application. METHODS: Fifty-seven patients (83 eyes) diagnosed as pterygium complicated with conjunctivochalasis, at our hospital from July 2011 to June 2012, were selected. Patients were treated with pterygium excision and conjunctival autotransplantation combined with sclera fixation surgery, then therapeutic bandage contact lenses were applied. The efficacy of simultaneous surgery was evaluated based on vision changes, tear dynamics, and other complications. Histopathological changes were investigated on removed bulbar conjunctival tissue, using hematoxylin eosin (HE) and Masson's trichrome staining. RESULTS: (1) Three months after the operation, the success of simultaneous surgery in the treatment of pterygium was 97.6 %, and the recurrence was 2.4 %. Based on subjective evaluation, the success of the simultaneous treatment of conjunctivochalasis was 95.2 %, and failure was 4.8 %. Based on objective evaluation, the success rate was 94.0 % and the recurrence rate was 6.0 %. (2) Visual acuity of the 83 eyes was significantly improved after surgery, and was statistically significant (X 2 = 10.29, P < 0.05). (3) Three months after surgery, the height and integrity of the tear meniscus, tear film break-up time, and chloramphenicol test results of the 83 eyes were significantly improved and there was a statistically significant difference (X 2 the height and integrity of tear meniscus = 147.24, X 2 tear film break-up time = 81.17, X 2 chloramphenicol test = 17.41, P < 0.01). (4) Complications after the operation such as granulation hyperplasia, constrictive fornix, oculomotor defect, and other complications were not observed. (5) Pathological observations, using HE and Masson's trichrome staining of removed bulbar conjunctival tissue, showed several pathological changes, including obvious squamous epithelial hyperplasia, parakeratosis, basal cell pigmentation, lamina propria hemorrhage, infiltration of lymphocytes, and reduction of elastic fibers and collagen fibers. CONCLUSION: Pterygium excision and conjunctival autotransplantation, combined with sclera fixation followed by therapeutic contact lens use was safe, effective, and suitable for simultaneous treatment of pterygium complicated with conjunctivochalasis.

Concentration change of TGF -ß 1 in aqueous humor of rabbits.

OBJECTIVE: To observe the influence of the the transforming growth factor ß1 (TGF-ß1) eye drops on rabbit aqueous humor TGF-ß1 concentration, and to analyze the best drug concentration. METHODS: A total of 30 New Zealand white rabbits were randomly divided into 5 groups with 6 in each. Rabits in control group had PBS eye drops, group A, B, C, D adopted TGF-ß1 eye drops at 0.5, 1.0, 2.0, 4.0 mg/L, respectively, 4 times a day. Aqueous humor of right eye was extracted 1 week after administration to detect concentration changes of TGF-ß1 by ELISA; rabbits in fpur hroups adopted 2.0 mg/L eye drops to left eyes 4 times a day, 0.2 mL aqueous humor was extracted left eye at the scheduled time point 0, 30 min, 2 h, 4 h, 24 h for testing, the slit lamp was used to observe the cornea, chamber and lens. RESULTS: No obvious pathological changes in conjunctiva, cornea, rabbit conjunctival, anterior chamber, and the lens was found. Concentration of TGF-ß1 in rabbit aqueous humor in C, D group was significantly higher than the control group (P<0.05). CONCLUSIONS: TGF-ß1 eye drops at 2.0 mg/L, 4.0 mg/L can significantly increase concentration of TGF-ß1 in rabbit aqueous humor, withe good ocular surfac permeability.

Spontaneous regression of retinopathy of prematurity: incidence and predictive factors.

AIM: To evaluate the incidence of spontaneous regression of changes in the retina and vitreous in active stage of retinopathy of prematurity(ROP) and identify the possible relative factors during the regression. METHODS: This was a retrospective, hospital-based study. The study consisted of 39 premature infants with mild ROP showed spontaneous regression (Group A) and 17 with severe ROP who had been treated before naturally involuting (Group B) from August 2008 through May 2011. Data on gender, single or multiple pregnancy, gestational age, birth weight, weight gain from birth to the sixth week of life, use of oxygen in mechanical ventilation, total duration of oxygen inhalation, surfactant given or not, need for and times of blood transfusion, 1,5,10-min Apgar score, presence of bacterial or fungal or combined infection, hyaline membrane disease (HMD), patent ductus arteriosus (PDA), duration of stay in the neonatal intensive care unit (NICU) and duration of ROP were recorded. RESULTS: The incidence of spontaneous regression of ROP with stage 1 was 86.7%, and with stage 2, stage 3 was 57.1%, 5.9%, respectively. With changes in zone III regression was detected 100%, in zone II 46.2% and in zone I 0%. The mean duration of ROP in spontaneous regression group was 5.65±3.14 weeks, lower than that of the treated ROP group (7.34±4.33 weeks), but this difference was not statistically significant (P=0.201). GA, 1min Apgar score, 5min Apgar score, duration of NICU stay, postnatal age of initial screening and oxygen therapy longer than 10 days were significant predictive factors for the spontaneous regression of ROP (P<0.05). Retinal hemorrhage was the only independent predictive factor the spontaneous regression of ROP (OR 0.030, 95%CI 0.001-0.775, P=0.035). CONCLUSION: This study showed most stage 1 and 2 ROP and changes in zone III can spontaneously regression in the end. Retinal hemorrhage is weakly inversely associated with the spontaneous regression.

Intraoperative mitomycin C versus intraoperative 5-fluorouracil for trabeculectomy: a systematic review and meta-analysis.

PURPOSE: The aim of this study was to assess the intraoperative application of mitomycin C (MMC) compared to 5-fluorouracil (5-FU) on the outcome of trabeculectomy and to examine the balance of risk and benefit. METHODS: Pertinent studies were selected through systematic searches of major literature databases, including the Cochrane Library, PubMed, Embase, and Chinese Biomedicine Database. Internet searches of search engines, the professional associations' websites, and the manufacturers' databases were also performed. Clinical controlled trials comparing 5-FU with MMC in trabeculectomy were selected. The primary efficacy measure was the weighted mean difference (WMD) in percentage intraocular pressure reduction (IOPR%) at follow-up end point. The secondary efficacy measure was the relative risk (RR) for "qualified" (with or without medical therapy) success of trabeculectomy at follow-up end point. The third efficacy measure was RR for "complete" (without medical therapy) success of trabeculectomy at follow-up end point. The fourth efficacy measure was RR for adverse events, including wound leak, hypotony, endophalmitis, and shallow anterior chamber (AC). The pooled effects were calculated using the random effects model by RevMan version 5.0 software. RESULTS: Eight studies enrolling a total of 536 patients were included in the meta-analysis. MMC was associated with significantly more IOPR% compared with 5-FU, with a WMD of 7.09 [95% confidence interval (CI) 1.47-12.70] at follow-up end point (P=0.01). MMC was comparable with 5-FU in qualified success rate, with a RR of 1.09 (0.99-1.20) at follow-up end point (P=0.09). MMC was comparable with 5-FU in complete success rate, with a RR of 1.17 (0.79- 1.75) at follow-up end point (P=0.43). Rates of adverse events did not differ significantly between 5-FU and MMC, with an RR of 0.71 (0.22-2.28) for bleb leakage, 1.40 (0.72-2.72) for hypotony, 1.63 (0.27-9.75) for endophthalmitis, and 0.95 (0.41-2.21) for shallow AC. CONCLUSIONS: Intraoperative MMC is more effective in lowering IOP in trabeculectomy compared with intraoperative 5-FU, but is comparable with intraoperative 5-FU in both qualified and complete success rate. Intraoperative use of both agents may contribute equally to adverse events.

Blockade of toll-like receptor 2 expression and membrane translocation in rat corneal epithelial cells by glucocorticoid (TobraDex) after penetrating keratoplasty.

PURPOSE: To evaluate the role of Toll-like receptor (TLR) 2 and the effect of glucocorticoid on immune rejection of penetrating keratoplasty. METHODS: Allograft corneal transplantation was performed between host Sprague Dawley (SD) and Wistar donor rats. The expression of TLR2 messenger RNA (mRNA) and protein in corneas was determined by reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemistry, and immunofluorescence on days 5, 7, and 9 after operation. Three groups were included: allograft, allograft treated with TobraDex (Alcon, Rijksweg, Belgium), and isograft. Normal rat corneas were included as an additional control. RESULTS: Various degrees of congregation of inflammatory cells and neovascularization of grafts were confirmed by histopathology. Immunohistochemistry revealed that TLR2 was expressed in epithelial, stromal, and endothelial cells of normal tissue, and in all of the grafts. Immunofluorescence analysis of TLR2 showed membrane staining of epithelial cells in the allografts on days 7 and 9. This was absent in the isografts and the allografts treated with TobraDex. TLR2 mRNA was detected in normal corneas, and levels were increased in all of the grafts, as determined by quantitative reverse transcription-polymerase chain reaction. By day 9 after transplantation, a 3.6-fold increase in TLR2 mRNA was observed in the allografts compared with the isografts or the allografts treated with TobraDex, which was statistically significant, at P < 0.005. CONCLUSIONS: Expression of TLR2 in the rat cornea was significantly increased and concurred with the allograft rejection, but was effectively blocked by treatment with TobraDex.

Topical dihydroartemisinin inhibits suture-induced neovascularization in rat corneas through ERK1/2 and p38 pathways.

AIM: To determine if topical instillation of dihydroartemisinin (DHA) inhibits corneal neovascularization (NV) in rats and to investigate the role of the extracellular regulated kinases (ERK) 1/2 and p38 pathways in this process. METHODS: Suture-induced corneal NV was produced in rats and the eyes were topically treated with different concentrations of DHA (20mg/L, 10mg/L or 5mg/L) or normal saline 4 times a day for 7 days. The corneal NV was quantified as the proportion of NV area to the whole cornea. Western blot was used to determine the expressions of vascular endothelial growth factor (VEGF) and the phosphorylation status of VEGF receptor-2, ERK1/2 and p38 in the corneas. Immunofluorescent staining was used to determine the expressions of phospho-ERK1/2 and phospho-p38 in the corneal tissues from the eyes treated with 20 mg/L DHA (DHA group) or normal saline (control group). RESULTS: The proportion of corneal NV area in the eyes treated with normal saline or DHA at dosages of 20mg/L, 10mg/L or 5mg/L was (23.74±3.00)%, (15.73±2.88)%, (19.53±2.42)%, and (23.38±2.79)%, respectively. In the eyes treated with 20mg/L or 10mg/L DHA, the corneal NV area was significantly reduced when compared to that in eyes with normal saline (P<0.05). Western blot analyses revealed that 20mg/L DHA significantly inhibited the expressions of VEGF and phospho-VEGFR-2. Both 20mg/L and 10mg/L DHA inhibited the expressions of phospho-ERK1/2 and phospho-p38. Immunofluorescent staining further demonstrated that 20mg/L DHA lowered the expression levels of phospho-ERK1/2 and phospho-p38 in the corneas with suture-induced NV. CONCLUSION: Suture-induced NV in rat corneas was significantly inhibited by topical treatment with 20mg/L and 10mg/L DHA. The results suggest that the effects could be partially dependent on the DHA-mediated inhibitions of the ERK1/2 and p38 pathways.

Proteomic analysis of human serum from diabetic retinopathy.

AIM: To establish and compare serum proteomic of diabetic retinopathy (DR) patients in various phases and discuss pathogenesis of DR so as to find out possible serum specific molecular markers for early diagnosis of DR. METHODS: Thirty-two subjects were divided into four groups: one group of eight type 2 diabetes mellitus (T2DM) patients without apparent DR (No-DR, NDR), one group of eight T2DM patients with non-proliferative diabetic retinopathy (NPDR), one group of eight T2DM patients with proliferative diabetic retinopathy (PDR) and one group of eight healthy volunteer participants. Two dimensional fluorescence difference gel electrophoresis (2D-DIGE) was applied to establish differential protein expression profiles in four groups. Matrix-assisted laser desorption/ionization time of flight tandem mass spectrometry (MALDI-TOF-TOF MS) was applied to identify mass spectrometry of differential proteins and analyze follow-up bioinformatics. RESULTS: 2D-DIGE maps of serum protein were satisfactory obtained from NDR, NPDR, PDR and normal control groups. Twenty-six different proteins spots were screened (the volume ratio was >1.5 based on DeCyder software analysis). Twenty-four of them were verified and two of them were not. Fifteen proteins were verified. Most of them were high-abundant proteins in serum. The four relatively low-abundant ones were beta 2-glycoprotein I (ß(2)-GPI), alpha2-HS-glycoprotein(AHSG), alpha1-acid glycoprotein(α(1)-AGP) and apolipoprotein A-1(apo A-1). ß(2)-GPI expression was gradually increased in the development of DR but unrelated to the severity of DR. The volume ratio of ß(2)-GPI is 1.54, 2.43, and 2.84 in NDR, NPDR and PDR group respectively compared with normal control group. CONCLUSION: Serum proteomic analysis of 2D-DIGE combined with MALDI-TOF-TOF MS is feasible to be applied in the study of DR. ß(2)-GPI probably takes part in the process of DR occurrence and development and it could be a candidate biomarker on DR diagnosis in early phase.

[Pathological changes of the cornea in rabbits with hyphema and concurrent ocular hypertension].

OBJECTIVE: To evaluate the impact of hyphema secondary to high intraocular pressure on corneal pathology in rabbits. METHODS: Thirty adult New Zealand rabbit were randomized into 3 equal groups, and in each rabbit, one eye served as the experimental eye with the other as the control eye. In the experimental eye, autoblood was injected into the anterior chamber to induce high intraocular pressure maintained for 3, 5, or 8 days. Only saline was injected into the control eye. After the injections, the cornea was observed with slit-lamp microscopy, and at 3, 5, or 8 days, the experimental and control eyes were taken from the 3 groups for microscopic examination of the corneas to detect the occurrence of cornea bloodstain with prolonged high intraocular pressure. Corneal edema, elastic fibers changes, growth of new blood vessels, changes of eosinophils, fibroblasts, lymphocytes and plasma cells, as well as the pathological changes of the corneal layers were observed and compared between the experimental and control eyes. RESULTS: Maintenance of high intraocular pressure for 8 days resulted in the most severe corneal edema and thickening, and histopathologically, the corneal stroma showed widened space between the elastic fibers and obvious fiber distortion. Neovascularization was seen in the marginal cornea where eosinophil infiltration occurred with a small number of lymphocytes, plasma cells and fiber cells. All the three groups showed more obvious edema in the posterior than in the anterior cornea. CONCLUSION: Prolonged hyphema with ocular hypertension results in aggravation of corneal edema, and corneal blood staining does not occur until 8 days of high intraocular pressure but corneal elastic fiber disruption can be seen, suggesting the impending irreversible pathological changes of cornea.

[Role of Toll-like receptor 2 in corneal graft rejection following penetrating keratoplasty].

OBJECTIVE: To gain insight into the role of Toll-like receptor 2 (TLR2) in graft rejection following penetrating keratoplasty, and investigate the expression of TLR2 mRNA in the corneal graft. METHODS: Penetrating keratoplasty was performed in 3 groups of rats for orthotopic autologous corneal transplantation (group A), allograft corneal transplantation (group B), or allograft corneal transplantation with hormone treatment (C). The transparency and neovascularization of the cornea were observed using a slit-lamp microscope and scored according to the rejection index, with normal cornea serving as the control. The corneal tissues were sampled at 5, 7, and 9 days after the transplantation for histopathological examination and detection of TLR2 mRNA expression using RT-PCR. RESULTS: With the passage of time, edema, opacities and neovascularization of the corneal graft occurred after the operation in all the groups. Seven days after the operation, the rejection index of group B, but not that of groups A and C, met the diagnostic criteria for graft rejection with also support by histopathological evidence. The expression of TLR2 mRNA was detected in normal corneas and augmented in the corneal grafts in the 3 transplantation groups. TLR2 mRNA expression in group B was significantly higher than that of group A, and the expression in group C decreased significantly in comparison with that in group B (P<0.05). CONCLUSION: As the recognition receptors of native immune system, TLR2 in the rejected corneal grafts may recognize the allograft antigen and play a role in acute graft rejection after penetrating keratoplasty.

[Effects of anti-CD25 monoclonal antibody on the level of cytokines in aqueous humour after rat penetrating keratoplasty].

OBJECTIVE: To investigate the toxicity of anti-CD25 monoclonal antibody (mAb) to rat cornea and its effects on the cytokines in the aqueous humour after penetrating keratoplasty (PKP), thereby evaluating the effect of anti-CD25 mAb in preventing corneal allograft rejection. METHODS: The corneal toxicity of anti-CD25 mAb at 50, 100 and 200 microg administered via subconjunctival injection was evaluated in 12 SD rats by histological examination and transmission electron microscopy. Another 93 SD rats were randomized into 5 groups, and transplantation of corneal allograft from Wistar rats was performed in 4 groups with the other group as the normal control. The 4 allograft groups were treated with saline, 100 microg anti-CD25 mAb, 100 microg anti-CD25 mAb with 50 microg dexamethasone, and 50 microg dexamethasone, respectively. The graft rejection was observed, the aqueous humour levels of IFN-gamma and IL-4 were measured with ELISA, and IFN-gamma mRNA expressions in the grafts detected with RT-PCR. RESULTS: anti-CD25 mAb at 50 or 100 microg did not show significant toxicity on the cornea, but at 200 microg, the mAb caused swelling of the corneal stromal cells and endothelial cells. After corneal allograft transplantation, a significant delay in allograft rejection was observed in the 3 groups with mAb or dexamethasone treatment as compared with that in saline group (P<0.05). IFN-gamma mRNA expression in the allograft on days 11 after PKP and in the aqueous humour on days 6 and 11 was markedly increased in saline group compared with that in the 3 treatment groups (P<0.05). The mean IL-4 level in the aqueous humour was significantly higher in the mAb group than in saline group (P<0.05), but markedly lower in anti-CD25 mAb+dexamethasone and dexamethasone groups than in anti-CD25 mAb group (P<0.05). CONCLUSIONS: Anti-CD25 mAb at 20 and 100 microg does not obviously affect the rat corneas. Anti-CD25 mAb inhibits IFN-gamma expression and promotes IL-4 the expression to reduce corneal allograft rejection, whereas anti-CD25 mAb with low-dose dexamethasone inhibits both IFN-gamma and IL-4 expressions to more effectively promote the graft survival.

[Recombinant type 1 adeno-associated virus-mediated transfection of ECV304 cells with enhanced green fluorescent protein gene in vitro].

OBJECTIVE: To assess the feasibility of recombinant type 1 adeno-associated virus (rAAV1) as a vector for gene therapy of corneal neovascularization. METHODS: The rAAV1 vector carrying enhanced green fluorescence protein (EGFP) gene (rAAV1-EGFP) was transfected into ECV304 cells at different multiplicities of infection (MOI=5 x 10(3), 5 x 10(4), 5 x 10(5)). EGFP expression in the cells was observed under inverted fluorescence microscope, and the EGFP-positive cell percentage determined by flow cytometry. MTT assay was used to assess the proliferation of the transfected cells. RESULTS: The cells with rAAV1-EGFP transfection at MOI of 5 x 10(5) began to exhibit GFP expression 2 days after transfection, and the fluorescence intensity increased with the MOI used for transfection. GFP expression reached the maximum on day 7, at the point of which the transduction efficiency of rAAV1-EGFP in ECV304 cells was 45.90%, 58.56% and 68.31% corresponding to MOIs of 5 x 10(3), 5 x 10(4), and 5 x 10(5), respectively. MTT assay did not reveal significant difference in the absorbance between the transfected cells and the control cells at 72 and 96 h after transfection. CONCLUSION: arAAV1-EGFP gene can be stably and efficiently expressed in ECV304 cells without causing cell growth inhibition, suggesting the potential of rAAV1 as a safe and efficient vector for gene therapy of corneal neovascularization.

[Role of dendritic cells in graft rejection after penetrating keratoplasty].

OBJECTIVE: To gain insight into the role of dendritic cells in graft rejection following penetrating keratoplasty by investigating their distribution in rat cornea. METHODS: Orthotopical corneal transplantation was performed and immunohistochemical staining of the whole-mount cornea and the spleen tissue specimen employed to determine the distribution of the dendritic cells in the cornea. RESULTS: Graft rejection occurred in all rats following the transplantation. No OX-62(+) dendritic cells were found in normal cornea but they were present in the epithelium of the cornea graft with allograft rejection. CONCLUSION: OX-62(+) dendritic cells presenting in the rejected cornea may be related to acute graft rejection after penetrating keratoplasty.