ObjectiveTo analyze the sensitivity and specificity of polymerase chain reaction (PCR) tests in the detection of cytomegalovirus (CMV) in the diagnosis of patients with acquired immune deficiency syndrome (AIDS), using aqueous humor samples. Methods25 AIDS patients (including 21 men and 4 women) were studied. The age of the patients varied from 24 to 59 years, with an average of (39.2±9.3) years. The CD4+ T cell count was from 1 to 523 cells/μl, with a medium of 40 cells/μl. They were infected with human immunodeficiency virus(HIV)for a period from 15 days to 9 years with a median of 10 months. They were divided into three groups according to the fundus and treatment, including untreated cytomegalovirus retinitis (CMVR), treated CMVR and control group. There were 10 patients without anti-CMV treatment and 7 patients treated previously with foscarnet or ganciclovir whose eyes were diagnosed CMVR. Control group has 8 patients who had normal fundus or minor retinopathy excluded from CMVR. Approximately 100 μl of aqueous humor was obtained by anterior-chamber paracentesis and PCR was performed in all cases. ResultsThere were CMV DNA in 9 of 10 eyes with untreated CMVR (90.0% sensitivity). Of 7 specimens from eyes with treated CMVR, 3 were CMV PCR positive (42.9% sensitivity). All 8 samples of the control group were negative for CMV DNA, indicating the clinical specificity of our PCR was greater than 99.9% for CMVR. The anterior chamber paracentesis did not cause any complications in our patients except for a patient with subconjunctival hemorrhage. ConclusionsThe assay had an estimated sensitivity of 90.0% in detecting untreated CMVR and a sensitivity of 42.9% in detecting CMVR that had been treated. The specificity of this assay was greater than 99.9%.
ObjectiveTo characterize proteomic profile in aqueous humor of patients with pathologic myopia (PM) using quantitative proteomic analysis, which may provide new clues to understand the mechanisms and possible treatments of PM.MethodsA cross-sectional study. From January 2019 to August 2019, aqueous humor samples (32 cataract patients) were collected for quantitative proteomic analysis using liquid chromatography tandem mass spectrometry at Tianjin Medical University Eye Hospital. There were 11 males and 21 females. They were 58-76 years old with an average age of 68.41±6.09 years old. Sixteen patients with PM were regarded as PM group, 16 patients without myopia were regarded as the control group. The aqueous humor samples (100-150 μl ) were collected from all patients before cataract surgery. Using protein quantification and non-labeled liquid chromatography tandem mass spectrometry analysis, differentially expressed proteins were obtained. Five different proteins were randomly selected for ELISA verification. The differentially expressed proteins were further analyzed by gene ontology enrichment and Kyoto Encyclopedia of Genes and Genomes, which were validated using ELISA in the other twenty samples of each group.ResultsA total of 583 proteins were identified and 101 proteins were found to be differentially expressed, including 63 up-regulated proteins and 38 down-regulated proteins. ELISA verification results showed that the expression trend of the 5 differentially expressed proteins between the PM group and the control group was consistent with the results of Label-free quantitative proteomics analysis. The main classifications of these differentially expressed proteins were protein-binding activity modulator, defense/immunity protein, protein modifying enzyme, metabolite interconversion enzyme, extracellular matrix protein, transfer/carrier protein and so on. The bioinformatics analysis suggested that PM was closely associated with inflammation and immune interactions, and remodeling of extracellular matrix.ConclusionsCompared with the control group, the protein expression profile of PM patients' aqueous humor specimens has obvious changes. These differences indicate that PM is closely related to inflammation and immune interaction and extracellular matrix remodeling.
Objective To observe the expression of vascular endothelial growth factor (VEGF) in aqueous humor and vitreous body in eyes with proliferative vitreo-retinal diseases, and to investigate the role of VEGF plays in the pathoge nesis of proliferative vitreo-retinal diseases. Methods The concentration of VEGF in aqueous humor and vitreous body in eyes with proliferative vitreoretinopathy (PVR), retinal vein occlusion (RVO), proliferative diabetic retinopathy (PDR), and neovascular glaucoma (NVG) were measured by double antibodies sandwich enzyme-linked immunosorbent assay (ELISA). Results The concentration of VEGF in aqueous humor and vitreous body in eyes with PVR, RVO, PDR and NVG were obviously higher than that in the control group (Plt;0.05), respectively. Among all of the diseases, the concentration of VEGF in aqueous humor and vitreous body decreased orderly in NVG, PDR, RVO and PVR (Plt;0.05). The concentration of VEGF in vitreous body in eyes with PVR, RVO, PDR and in the control group were much higher than that in aqueous humor in corresponding groups (Plt;0.05). There was a negative correlation between the disease history and content of VEGF in aqueous humor and vitreous body in patients with PVR (r=－0.819, －0.823；Plt;0.05). The disease history positi vely correlated with the concentration of VEGF in aqueous humor and vitreous body in patients with RVO (r=0.913, 0.929；Plt;0.05), and the time of vitreous hemorrhage positively correlated with the concentration of VEGF in aqueous humor and vitreous body in patients with PDR (r=0.905, 0.920；Plt;0.05). Conclusion The concentration of VEGF in aqueous humor and vitreous body in patients with proliferative vitreo-retinal diseases significantly increases, and VEGF may play an important role in the pathoge nesis of proliferative vitreo-retinal diseases. (Chin J Ocul Fundus Dis, 2006, 22: 313-316)
Objective To detect the changes of function of blood-aqueous barrier in different Syndrome stages of patients with Vogt-Koyanagi-Harada (VKH) syndrome in order to provide the appropriate therapy. Methods According to clinical manifestation, 77 patients (144 eyes) with VKH syndrome were divided into 4 groups: 10 cases in posterior uvietis stage group (20 eyes), 27 in anterior uveal involvement stage group (50 eyes), 23 in recurrent anterior uvitis stage group (41 eyes), and 17 in convalescent stage group (33 eyes). The other 50 cases (100 eyes) were in the control group. Flare and cells of anterior chamber in patient with VKH Syndrome at different stages were graded and measured by laser flare and cell meter (LFCM) and slitlamp microscope. Results According to the results of slitlamp biomicroscopy, anterior chamber flare and cells were at the 0 grade in the patients at posterior uvietis stage (20 eyes). The results of LFCM examination revealed that the flare value and cells were (9.7±3.4) pc/ms and (0.9±0.6)/0.5 mm3 in posterior uvietis stage group, and (5.3±2.3) pc/ms and (0.8±0.6)/0.5 mm3 in the control group. The differences between the two groups were significant (Plt;0.001) and insignificant (P=0.899), respectively. In anterior uveal involvement stage group, the cells in anterior chamber was at grade 1+ in 25 eyes, 2+ in 19, and 3+ in 6, respectively, while the flare was at grade 1+in 27 eyes and 2+ in 23; the number of cells in anterior chamber was (13.7±6.5)/0.5 mm3，(40.8±17.6)/0.5 mm3, and (75.7±25.5)/ 0.5 mm3 respectively, and the value of flare was (31.4±12.8) pc/ms and (133.4±59.5) pc/ms. In recurrent anterior uvitis stage group, the cells in anterior chamber was at grade 1+ in 19 eyes, 2+ in 15, and 3+ in 7, respectively, while the flare was at grade 1+ in 24 eyes and 2+ in 17; the number of cells in anterior chamber was (11.2±5.4)/0.5 mm3，(29.6±14.4 )/0.5 mm3,and (69.3±22.2)/0.5 mm3, respectively, and the value of flare was (34.94±14.3) pc/ms and (150.9±83.3) pc/ms. The flare and cells in anterior chamber both in anterior uveal involvement stage and recurrent anterior uvitis stage group were higher than that in the control group (Plt;0.001). In convalescent stage group, the cells was at grade 0 in 33 eyes and the flare was at grade 0 in 15 eyes and 1+ in 18; while the number of cells was (1.0±0.7)/0.5 mm3 which was insignificantly differed from that in the control group (P=0.310), and the value of flare was (9.5±4.8) pc/ms and (30.0±12.3) pc/ms which were both higher than that in the control group (Plt;0.001). Conclusions The breakdown of blood-aqueous barrier with different degrees occurs at each stage in VKH syndrome, whereas inflammatory cells appearing in anterior chamber are only noted at some certain stages. This is very significant to offer directional and effective treatment to the patients with VKH syndrome. (Chin J Ocul Fundus Dis, 2005, 21: 363-366)
Objective To detect the concentration of vascular endothelial growth factor (VEGF) in plasma and intraocular liquid (aqueous humor and vitreous body) in patients with deabetic retinopathy (DR) and the role VEGF plays in the development of DR. Methods The concentrations of VEGF in plasma, aqueous humor and vitreous body in DR and normal group were detected by ELISA. Results The concentration of VEGF in plasma was (34.47plusmn;1.76) pg/ml in non-DR group, (53.93plusmn;3.08) pg/ml in single DR group, (53.36plusmn;3.28) pg/ml in proliferative DR group, and (178.30plusmn;10.13) pg/ml in control group. There was no significant difference in the normal and the experimental groups (P＜0.05). The concentration of VEGF in aqueous humor was (184.8plusmn;12.60) pg/ml in proliferative DR group and (90.06plusmn;8.32) pg/ml in the control group, and there was significant difference between them (P＜0.05). The concentration of VEGF in vitreous body was (741.70plusmn;92.02) pg/ml in proliferative DR group and (94.38plusmn;21.21) pg/ml in the control group, and there was significantdifference between them (P＜0.05). There was no correlation of VEGF concentration in plasma and that in aqueous humor and vitreous respectively(P＞0.05), and positive correlation of VEGF concentration was found in vitreous body and HbA1c (r=0.9067,P＜0.01). Conclusions Concentration of VEGF in plasma in patients with DR is lower than that in the normal persons,but not correlated with the concentration of VEGF in aqueous humor and vitreousbody. The concentration of VEGF in aqueous humor and vitreous body increase in patients with proliferative DR, and the increase in vitreous body and the value of HbA1c of the patients correlate. (Chin J Ocul Fundus Dis，2004，20：343-345)
Objective To review the distribution and shifting trends of cultured bacteria from the aqueous humor and the vitreous body. Methods A retrospective analysis on distribution of Gram′s stain, the distribution and change of isolates was performed in 522 specimens (aqueous humor,261 and vitreous body,261) of patients with suspected endophthalmitis during a 10-year period (1989-1998). Results The positive cultures were 119 (aqueous humor,44 and vitreous body,75) of 522 specimens. The average positive rate was 22.8%. Gram-positive cocci constituting 45.4%(54) of total isolates followed by Gram-negative bacilli,34.5%(41);Gram-positive bacilli, 20.2%(24). In the positive bacterial cultures, enterobacteriaceae was the most common isolate, 18.5%, and the next was micrococcus, 16.0%; coagulase-negative staphylococcus,12.6%; and pseudomonas,10.9%.Comparing the data from 1989 through 1993 with the data from 1994 through 1998, the frequency of Gram-positive cocci had no significant change, while the frequency of Gram-positive bacilli was decreased and the percentage of Gram′s-negative bacilli was increased. Conclusions Gram-positive cocci and Gram-negative bacilli are the predominant pathogens of bacterial endophthalmitis. The percentage of Gram′s-negative bacilli has increased for 5 years. It is very important to comprehend the distribution and shifting trends of these pathogenic bacteria for diagnosis, prevention and treatment of bacterial endophthalmitis. (Chin J Ocul Fundus Dis, 2002, 18: 104-105)