[Clinical research of anterior chamber inflammation after phacoemulsification surgery in patients with complicated cataract secondary to uveitis].

OBJECTIVE: To investigate anterior chamber inflammation after phacoemulsification with intraocular lens (IOL) implantation in patients with Vogt-Koyanagi-Harada (VKH) syndrome or Behçet's disease (BD). METHODS: Cohort study. Seventeen patients (20 eyes) with complicated cataracts and VKH syndrome or BD who underwent phacoemulsification with IOL implantation at Zhongshan Ophthalmic Center, Sun Yat-Sen University (SYSU) between January 2010 and June 2011 were included as the experimental group in this study. Cataract surgery was performed on these patients only when uveitis had been under control for more than three months. Thirty patients (40 eyes) with age-related cataracts who underwent phacoemulsification with IOL implantation in the same period were included as the control group. Quantitative measurements of anterior chamber aqueous flare and inflammatory cells were conducted preoperatively and postoperatively using a Laser Flare Cell Meter (LFCM). Independent t-test was used to compare patients' ages, and the energy and time of phacoemulsification between the two groups. The Student's t-test was used to assess the differences between paired data preoperatively and postoperatively. Independent t-test was also used to assess the quantitative data between groups. RESULTS: The study recruited 20 eyes in the experimental group and 40 eyes in the control group, including 11 eyes from 9 VKH patients and 9 eyes from 8 BD patients. The preoperative and postoperative flare values in the experimental group were (19.86 ± 6.47), (44.28 ± 18.47), (35.60 ± 12.65), (23.85 ± 8.41), and (13.86 ± 4.27) pc/ms, respectively, which were statistically higher than that of the control group preoperatively, and on days 1, 7, 30, and 90 after surgery (tpre = 4.643, Ppre < 0.01; t1 = 6.035, P1 < 0.01; t7 = 3.595, P7 = 0.001; t30 = 4.658, P30 < 0.01; t90 = 3.308, P90 = 0.002). Aqueous flare in Group A and Group B declined to preoperative levels on day 30 (t = 0.320, P = 0.753) and day 7 (t = 0.454, P = 0.653). For the experimental group, the inflammatory cell count on day 1 and 7 was (83.46 ± 27.08) and (27.56 ± 8.32) cells/0.5 mm(3), respectively, which was significantly higher than the preoperative level [(6.47 ± 3.56)cells/ 0.5 mm(3), t1 = 5.261, P1 < 0.01; t7 = 2.766, P7 = 0.012]. On days 30 and 90, the inflammatory cell count was (11.43 ± 4.81) and (4.82 ± 2.29) cells/0.5 mm(3), respectively, and there was no statistically significant difference in the inflammatory cell count compared with the preoperative level (t30 = 2.348, P30 = 0.042; t90 = 1.376, P90 = 0.186). For the control group, inflammatory cell count reduced to pre-operative level on day 7 (t7 = 2.464, P7 = 0.018). CONCLUSIONS: Anterior chamber inflammation reaches peak levels one week postoperatively in VKH and BD patients who receive phacoemulsification with IOL implantation. It takes three months for the inflammation to recede, and might take longer for complete restoration of the blood-aqueous barrier.

[The blood-aqueous barrier changes after laser peripheral iridotomy or surgery peripheral iridectomy].

OBJECTIVE: To investigate the ocular blood-aqueous barrier (BAB) alteration after laser peripheral iridotomy (LPI) or surgery peripheral iridectomy (SPI) in patients with primary chronic angle-closure glaucoma (PCACG). METHODS: This was a clinical randomized controlled trial. Sixty eyes of 60 subjects with early stage of PCACG were randomly received either LPI or SPI and followed up postoperatively at day 3, week 1, 2, 3, and 4. Aqueous flare in anterior chamber was measured by FC-2000 flare-cell photometry, intraocular pressure (IOP) measured by tonometer, central corneal endothelium cell counted by endothelioscopy, peripheral anterior synechiae (PAS) detected by gonioscopy. Data were analyzed by using two-way ANOVA for repeated measures, independent samples t-test, paired t-test, nonparametric test, and Spearman rank correlation test. RESULTS: On follow-ups of pre-operative and post-operative 3 days, 1 week (w), 2w, 3w and 4w respectively, the mean aqueous flare values for LPI group were (5.47 ± 1.09), (11.96 ± 3.07), (8.08 ± 2.18), (5.68 ± 0.83), (5.80 ± 1.00), (5.69 ± 1.12) PC/ms, and for SPI group were (5.43 ± 1.13), (8.44 ± 3.22), (6.42 ± 1.77), (5.35 ± 0.71), (5.53 ± 1.26), (5.45 ± 1.23) PC/ms. During post-operative 1w the flare values in both LPI and SPI groups were significantly higher than that on pre-operation (t = -12.753, -8.101, P < 0.05; t = -5.971, -3.870;P < 0.05) and LPI group had a significantly higher mean flare value than SPI group (t = 4.329, 3.231;P < 0.05). The IOP spike in LPI group was significantly (χ(2) = 5.079, 4.022, P < 0.05) higher than that in SPI group at week 1 of post-operation. Increased IOP was positively correlated with BAB damage (r = 0.899, 0.833; P < 0.05). The numbers of medications required to maintain IOP ≤ 21 mm Hg (1 mm Hg = 0.133 kPa) at week 4 of post-operation in LPI was significantly (Z = -1.984, P < 0.05) more than that in SPI group. There were no significant differences in central corneal endothelium cell count at week 1 (t = -0.696, 0.008) and in extension of PAS at week 4 (Z = -1.270, -1.490) of post-operation when compared to pre-operation (P > 0.05). No obvious complications occurred in both groups. CONCLUSIONS: Our results demonstrated that IOP spike in both of LPI and SPI is due, at least in part, to BAB damage, which appears to be more severe in LPI group and can recover within two weeks. PAS progression and central corneal endothelium cell loss are not aggravated in 1 month after operation.

[Expression of programmed death 1 and its ligands in iris-ciliary body from mice with anterior chamber-associated immune deviation].

OBJECTIVE: To study the expression and possible implication of programmed death 1 (PD-1) and its ligands in the iris-ciliary body from mice with anterior chamber-associated immune deviation (ACAID). METHODS: This was an experimental study. Twenty-four BALB/c mice were divided into ACAID group, negative controls, positive controls and phosphate-buffer saline controls. ACAID was evaluated by delayed-type hypersensitivity response. Expressions of PD-1 and its ligands (PD-L1, PD-L2) in the iris-ciliary body from ACAID mice were determined by real-time polymerase chain reaction and immunohistochemical studies. RESULTS: Delayed-type hypersensitivity was not detected in ACAID group, indicating that ACAID was induced successfully. The expression of PD-1 mRNA in ACAID group was higher than that in all other groups (F = 248. 109, P < 0.05). The positive controls showed a high expression of PD-L1 mRNA (F = 179. 033, P < 0.05). PD-1 and PD-L1 positive cells with round or oval shapes were found in whole iris-ciliary body, especially in the iris base and pupil margin. Expression of PD-1 and PD-L1 proteins were similar to their expression at mRNA levels. Neither mRNA nor protein of PD-L2 was detected in the iris-ciliary body in all groups. CONCLUSION: An increased expression of PD-1 in the iris-ciliary body of ACAID mice indicates that this is involved in the induction of ACAID.

Role of T-cell receptor V beta 8.3 peptide vaccine in the prevention of experimental autoimmune uveoretinitis.

BACKGROUND: T-cell receptor (TCR) plays an important role in the development of autoimmune diseases. Recently, it was reported that immunization of animals with TCR peptide derived from the pathogenic cells could prevent autoimmune diseases. The aim of this study was to investigate whether vaccination with a synthetic peptide from the hypervariable region of TCR V(beta) 8.3, an experimental autoimmune uveoretinitis (EAU)-associated gene, was able to prevent the disease. METHODS: EAU was induced in Lewis rats by immunization with IRBP R16 peptide emulsified in complete Freund's adjuvant (CFA). The clinical and histological appearances were scored. Delayed type hypersensitivity (DTH) and lymphocyte proliferation were detected. Cytokine levels of aqueous humour, supernatants of cells from spleen and draining lymph nodes were measured by enzyme linked immunosorbent assay (ELISA). Gene expression of TCR V(beta) 8.3 on CD(4)(+) T cells was examined by real time quantitative polymerase chain reaction (PCR). RESULTS: After vaccination, the intraocular inflammation was significantly mitigated, antigen specific DTH and lymphocyte proliferation responses were suppressed, interleukin (IL)-2 in aqueous humour, interferon (IFN)-gamma and IL-2 produced by the spleen and draining lymph node cells were significantly decreased, whereas the production of IL-4 and IL-10 were increased. The response of draining lymph node cells to TCR V(beta) 8.3 peptide was enhanced after vaccination. Inoculation with CFA alone did not affect the severity of EAU and the above parameters. The suppression of EAU was much stronger in the group of four fold inoculations than the group of two fold inoculations. The expression of TCR V(beta) 8.3 gene was significantly reduced in the group of fourfold inoculations. CONCLUSION: Vaccination with the synthetic TCR V(beta) 8.3 peptide could remarkably inhibit the development of EAU.

GATA-3 expression in the development of anterior chamber associated immune deviation.

BACKGROUND: Anterior chamber associated immune deviation (ACAID) is characterized by a Th2 cell response. GATA-3 has been shown to be necessary for the activation of Th2 cells. This study was designed to examine the expression of GATA-3 in the development of ACAID. METHODS: ACAID was induced by injection of 50 microg interphotoreceptor retinoid binding protein (IRBP) into the anterior chamber (AC) of Wistar rats. Delayed-type hypersensitivity (DTH) was evaluated on day 3, 7, 14, 21, 28 after IRBP inoculation. GATA-3 expression was detected using immunohistochemical staining. The expression of GATA-3 mRNA at different time points after AC injection of IRBP was assayed by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: A significant DTH reaction was observed in Wistar rats on day 3 and 5 after IRBP inoculation. The DTH reaction was decreased 7 days after IRBP inoculation. GATA-3 expression was weak at both mRNA and protein levels in the normal spleen, but was significantly increased on day 5, 7, 14, and 21 after AC injection of IRBP. CONCLUSION: The expression of GATA-3 is increased during ACAID, suggesting that GATA-3 may be involved in the development of ACAID.

[The inhibiting effects of suramin on bFGF induced proliferation of cultured human RPE cells].

OBJECTIVE: To study the inhibiting effects of suramin on FGF induced proliferation of cultured RPE cells and discuss the effects and mechanisms of suramin in the prevention of proliferative vitreoretinopathy (PVR). METHODS: Human RPE cells at passage 6 were seeded into 96-well plates. In the 5 control groups, different concentrations of bFGF (1, 10, 100 and 500 ng/ml) or suramin (31.25 microg/ml) were added into the culture medium. In the 4 treatment groups, 31.25 microg/ml suramin with different concentrations of bFGF (1, 10, 100 and 500 ng/ml) were added to the medium. The blank group was incubated with serum-free DMEM only. After incubated for 48 hours, the proliferation of cells in all groups was measured by the MTT assay and expressed as light absorption values. The separate and combine effects of suramin and bFGF were analysed. RESULTS: Under the phase-contrast microscope, no changes of cell morphology could be detected between various groups. Suramin showed an inhibitory effects on bFGF induced proliferation of RPE cells (F=6.73, P < 0.01). Light absorption value (A value) in bFGF-treated control groups was higher than that of the blank group and reached to the peak in cells treated with 10 ng/ml of bFGF; with no further increase in cells treated with higher levels of bFGF. The A value in suramin-treated control group was lower than that of the blank group. The A-value in all treated groups was lower than that of the control groups treated with various concentrations of bFGF. In the treated group, the highest A value was detected in cells treated with 100 ng/ml bFGF. With further increase of bFGF concentration to 500 ng/ml, the A-value was not statistically different from that of cells treated with 100 ng/ml bFGF. CONCLUSIONS: Suramin can inhibit the bFGF induced proliferation of RPE cells. The mechanisms of the effects of suramin in the inhibition of proliferation of RPE cells and in the prevention of PVR at least partly due to its competition effects with the growth factors.

[The clinical feature, diagnosis and treatment of uveitis associated with juvenile chronic arthritis].

OBJECTIVE: To investigate the clinical features, diagnosis and treatment of uveitis associated with juvenile chronic arthritis (JCA). METHODS: A retrospective study was performed on the clinical data of 26 patients with uveitis associated with JCA, referred to Zhongshan Ophthalmic Center from 1996 to 2002. Taking of history, examination with slit-lamp microscope and ophthalmoscope were carefully performed in all of these patients. Laboratory tests including antinuclear antibodies, erythrocyte sedimentation rate, rheumatoid factor, C reactive protein and antistreptolysin O were used to disclose possible causes. Human leukocyte antigen B 27 and X-ray examination of sacroiliac joints and spine were carried out if necessary. Data about the treatment, visual outcome and complications were analyzed. RESULTS: Twenty-six patients, 11 males and 15 females, were included in the present studies. Age of onset of JCA and uveitis averaged 8 years and 9 years, respectively. Twenty-one patients had chronic anterior uveitis. Acute anterior uveitis and chronic panuveitis were noted in 3 and 2 patients, respectively. Twenty-two patients had bilateral uveitis, the other 4 had unilateral involvement. The ophthalmologic examination revealed that 33 of the 48 affected eyes showed mild aqueous humor flare, 24 had few cells in the anterior chamber. Complicated cataract, band keratopathy and secondary glaucoma were noted in 30, 20 and 12 eyes, respectively. The laboratory examination revealed positive antinuclear antibodies in 18 cases and rheumatoid factor positive in one case. In acute episode, patients were treated with extensive topical cycloplegic agents and corticosteroids eyedrops. In the 6 cases with severe uveitis, 3 patients were treated with cyclosporine A (5 mg.kg(-1).d(-1)) and the other 3 were treated with chlorambucil (0.1 mg.kg(-1).d(-1)). The intraocular inflammation in all of these patients was satisfactorily controlled with these treatments. Visual acuity improvement was noted in 32 affected eyes but not in the other 16 eyes which had already serious complications before the treatment. CONCLUSIONS: Uveitis associated with JCA is characterized by a chronic and recurred iridocyclitis, which usually developed within 5 year after JCA onset. Although the intraocular inflammation associated with JCA is usually white uveitis, complications such as cataract, secondary glaucoma and keratopathy occur frequently in these patients. The diagnosis is mainly based on typical clinical manifestations, the history of arthritis and positive antinuclear antibodies. Adequate application of cycloplegic agents, corticosteroids eyedrops, immunosupressives and the regular follow-up should be kept in mind in the treatment of these patients.

[Clinical diagnosis and treatment of uveitis associated with ankylosing spondylitis].

OBJECTIVE: To investigate the clinical features, diagnosis, treatment and prognosis of uveitis associated with ankylosing spondylitis (AS). METHODS: A history about low back pain was carefully reviewed in patients with uveitis and X-ray examination was performed if necessary. Forty-four patients were diagnosed as AS from January 1996 to June 2001 in Zhongshan Ophthalmic Center according to the modified New York criteria for AS. Data concerning these patients, especially with respect to the clinical features, diagnosis and treatment, were retrospectively analyzed. RESULTS: Of 44 patients, 41 were male. The age of the patients at onset of uveitis was (33 +/- 15) years old. All of the 34 patients who had a uveitis history over 40 days showed a recurrent inflammation. In 18 patients (52.9%), the interval between the relapse of uveitis was more than one year. Bilateral ocular involvement was found in 23 out of 34 patients with recurrent uveitis. However, none of them showed a bilateral inflammation at the onset of uveitis. All patients showed acute anterior uveitis with a duration of (27 +/- 12) days. All patients had definitely radiological evidences of bilateral sacroiliitis, although not all of them had typical history of lower back pain. Treatment with corticosteroids eyedrops and cycloplegic agent was used in all patients. Vision equal to or better than 1.0 was achieved in 82.5% of these patients. CONCLUSIONS: Uveitis associated with AS is characterized by acute nongranulamatous anterior uveitis with recurrent episodes in male. Diagnosis is made according to typical clinical features and radiological evidences of bilateral sacroiliitis. Treatment with corticosteroid eyedrops and cycloplegic is able to resolve the inflammation rapidly and leads to a good prognosis in most patients.

[Preventive effect of anterior chamber associated immune deviation on endotoxin-induced uveitis].

OBJECTIVE: To determine the effect of anterior chamber associated immune deviation (ACAID) on endotoxin-induced uveitis (EIU) and the possible mechanism. METHODS: ACAID animal model was induced by injection of 5 microl IRBP (10 microg/microl) into the anterior chamber (AC) of Spar-Dawley (SD) rats. Then 200 microg LPS was injected into hind footpads at different time points after AC inoculation. The animals were randomly divided into 3 groups: positive control (LPS injection only), 3 d group (LPS injection 3 days after IRBP inoculation), 7 d group (LPS injection 7 days after IRBP inoculation). Delayed type hypersensitivity (DTH) was examined to determine the development of ACAID. Then the serum level of IL-10 was evaluated by ELISA, and GATA-3 expression at the different time points after IRBP injection was assayed by reverse transcription polymerase chain reaction (RT-PCR) and Western blot on mRNA and protein level respectively. The ocular inflammation was observed clinically; at the same time, the eye was extirpated and histological examination was performed. RESULTS: In control and 3 d groups, significant DTH reaction was induced, but the serum level of IL-10 could not be detected and GATA-3 expression was not increased. While in 7 d group, the DTH reaction could not be induced, and IL-10 and GATA-3 expression increased significantly at both the mRNA and protein levels. The clinical manifestation was significantly alleviated in the 7 d group; Histological examination displayed that the inflammatory cells were significantly reduced in iris/ciliary body, anterior and posterior chambers, vitreous body and retina of the rats in 7 d group. CONCLUSION: The development of ACAID can reduce the ocular inflammation induced by LPS, that is related to the increase of GATA-3 and IL-10 expression.

[Effect of endotoxin-induced uveitis on GATA-3 expression and ACAID development].

OBJECTIVE: To investigate the changes of the GATA-3 expression and development of anterior chamber associated immune deviation (ACAID) after anterior chamber (AC) injection of interphotoreceptor retinoid binding protein (IRBP) under ocular inflammation. METHODS: ACAID was induced by injection of IRBP into the AC of 30 Spar-Dawley (SD) rats. Then the animals were divided into -4 days group, -24 hours group, 0 hour group, 3 days group, and 7 days group according to the lipopolysaccharide (LPS) injection 4 days and 24 hours before, or 0 hour, 3 days and 7 days after IRBP inoculation respectively. 6 rats were used as controls 8 days after IRBP injection, Serum interleukin-4 (IL-4) was evaluated to count the development of ACAID; Western blot and RT-PCR were used to determine the protein and mRNA levels of GATA-3 expression. RESULTS: In -24 hours group and 0 hour group, the ocular inflammation reached a maximum 24 hours after LPS injection; on 8 days after IRBP inoculation, serum IL-4 couldn't be detected and GATA-3 expression were not changed both on mRNA and protein levels compared with control group. In -4 days group, the ocular inflammation was subsided gradually 24 hours and disappeared 96 hours after LPS injection; serum IL-4 and GATA-3 expression were significantly elevated 8 days after IRBP injection. In 7 days group, the serum IL-4 and GATA-3 expression in spleen increased 8 days after IRBP inoculation. CONCLUSION: In ocular inflammation, the up-regulation of GATA-3 expression is inhibited and ACAID development is blocked after antigen was injected into anterior chamber. Once GATA-3 is up-regulated, LPS injection cannot affect ACAID development.

[Increased expression of Fas and FasL mRNA in peripheral blood lymphocytes in Vogt-Koyanagi-Harada syndrome].

OBJECTIVE: To evaluate the expression of Fas and FasL messenger RNA (mRNA) in peripheral blood lymphocytes and its possible role in the pathogenesis of Vogt-Koyanagi-Harada syndrome. METHODS: Blood samples were obtained from 16 patients with Vogt-Koyanagi-Harada syndrome and 19 healthy individuals from January to June, 2000. Total RNA was extracted from peripheral blood lymphocytes. cDNA was prepared from RNA with oligo d(T)(18) and M-MuLV reverse transcriptase. The expression of Fas mRNA and FasL mRNA in peripheral blood lymphocytes was determined using fluorescent quantitative polymerase chain reaction (FQ-PCR). Amplification was carried out through 32 cycles, 30 seconds denaturation at 93 degrees C, 30 seconds annealing at 58 degrees C, and 45 seconds primer extension at 72 degrees C. The final extension time was 7 minutes. RESULTS: The expression of Fas mRNA on peripheral blood lymphocytes [(1.6 +/- 2.0) x 10(6)] was significantly higher than that of the controls [(5.7 +/- 2.0) x 10(5)] (t = 4.50, P < 0.05). The mRNA expression of FasL on peripheral blood lymphocytes [(1.8 +/- 1.5) x 10(6)] was also higher than that of the controls [(4.8 +/- 3.5) x 10(5)] (t = 9.57, P < 0.05). CONCLUSIONS: A high level of Fas mRNA and FasL mRNA expression in activated peripheral blood lymphocytes is observed in patients with Vogt-Koyanagi-Harada syndrome. These long-persisted activated lymphocytes may be responsible for the perpetuation and recurrence of Vogt-Koyanagi-Harada syndrome.

[Evaluation of the problems in the treatment of uveitis].

OBJECTIVE: To address the problems present in the treatment of uveitis. METHODS: A retrospective analysis was made on 154 patients (268 eyes) with different entities of uveitis, coming from all over China and referring to Zhongshan ophthalmic center from April to October of 2000. The data of these patients, especially with respect to the usage of corticosteroids and antibiotics in the referred hospitals, were analyzed. RESULTS: Corticosteroids drops, subconjunctival or retrobulbar injection and systemic corticosteroids were respectively used in 90.3% and 84.0% of these eyes, and 92.9% of these cases. Intravenous injection of corticosteroids was rather popular, accounting for 85.3% of the cases who were treated with systemic corticosteroids. Improperly aggressive treatment with corticosteroids or undue application of their eye drops was equally found in patients from county-, city- and province-level hospitals, especially in Behcet's disease, anterior uveitis and Fuchs heterochromic uveitis. Antibiotics drops, subconjunctival injection and systemic administration of antibiotics were respectively used in 97.0% and 71.6% of these eyes and 83.8% of these patients. Immunosuppressive drugs, other than the corticosteroids, were used in 6 patients only. CONCLUSIONS: Corticosteroids should be used in proper methods, dosage and duration based on the entities and severity of uveitis. It is not necessary to treat noninfectious uveitis with antibiotics.

[Quantitative determination of aqueous flare and cells in the eyes of patients with inflammation of anterior uvea].

OBJECTIVE: To quantify aqueous flare and cells in the eyes of patients with inflammation of anterior uvea by FC-2000 laser flare cell meter (LFCM), and to compare these results with those obtained with slit lamp microscopy. METHODS: Aqueous flare and cells of 194 eyes of 110 patients with inflammation of anterior uvea and 52 eyes of 52 healthy subjects were graded into 0, 1+, 2+, 3+ and 4 + scale based on a previously described system using slit lamp microscopy. LFCM was also used for evaluation of aqueous flare and cells. RESULTS: All eyes in normal individuals were graded as "0" scale of both flare and cells by silt lamp microscopy. Flare of grade 0, 1+ and 2+ were noted in 74, 98 and 18 eyes, and cell of grade 0, 1+, 2+, 3+ and 4+ were noted in 124, 26, 19, 14 and 11 eyes in uveitis patients, respectively. LFCM examination revealed that the mean flare values in uveitis eyes with flare of grade 0, 1+ and 2+ were 7.9, 29.5 and 189.0 photon count/ms, respectively. In patients with flare of grade 3+ and 4+, LFCM readings were unreliable because of increased background noise. There was significant correlation between slit lamp examination and the laser flare measurement for flare of grade 0, 1+ and 2+ (r = 0.75, P < 0.001). The mean flare values were significantly higher in patients with flare of grade 0, 1+ and 2+ than that in normal controls (5.3 pc/ms) (t = 5.872, P < 0.05). The mean cell numbers in the eyes with cell of grade 0, 1+, 2+, 3+ and 4+ were 1.5, 12.1, 33.9, 84.9 and 193.1 count/0.5 mm(3), respectively. The results of slit lamp examination showed significant correlation with laser cell counts measurement (r = 0.72, P < 0.001). The mean cell numbers were significantly higher in uveitis patients than that in normal controls (0.9 count/0.5 mm(3)) (t = 7.351, P < 0.05). CONCLUSIONS: Our results indicate that LFCM is able to evaluate precisely the mild and moderate breakdown of blood aqueous barrier and inflammation of the anterior uvea tract, therefore it provides an important parameter for the treatment of anterior uveitis.