The fundus lesions caused by high myopia (HM) often lead to irreversible visual impairment or even blindness. However, the pathogenesis of HM and its fundus lesions is still unclear, the intraocular fluid detection technology of micro samples has brought new prospects for the early diagnosis, monitoring and intervention of the fundus lesions. The molecules associated with HM are various and functionally diverse, intermolecular interactions are staggered and the specific mechanism is complex. With the development of intraocular fluid detection technology, while gradually revealing the role of each molecule in the pathogenesis of HM, it is expected to successfully assist clinical work in the future, providing outpost markers for the progress of myopia and targets for early intervention, or providing a new therapy choice for HM fundus lesions at the molecular level targeting pathogenesis, which is expected to provide more accurate and effective treatment for HM patients in the future.
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.
ObjectiveTo explore the predictive effect of intraocular fluid testing in the assessment of retinal detachment (RD) occurring in acute retinal necrosis syndrome (ARN). MethodsA retrospective study. From January 2019 to October 2023, 40 patients with 40 eyes diagnosed as ARN in Nanjing Medical University Eye Hospital were collected for this study. According to whether RD occurred during the follow-up period, the patients were divided into RD group (group A) and no RD group (group B), with 18 patients 18 eyes and 22 patients 22 eyes, respectively. All patients were given intravitreal 20 mg/ml ganciclovir 0.1 ml (ganciclovir 2 mg), 2 to 3 times per week after diagnosis. The concentrations of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (BFGF), vascular cell adhesion factor (VCAM), interleukin (IL)-6, IL-8, and IL-10 in the preaqueous solution were measured before the first injection of ganciclovir. The loads of herpes simplex virus (HSV) and varicella-herpes zoster virus (VZV) were detected by fluorescence quantitative polymerase chain reaction. Receiver operating characteristic curve (ROC curve) calculated and analyzed the area under ROC curve (AUC) of inflammatory cytokines in aqueous humor and HSV-DNA and VZV-DNA loads in predicting RD in ARN patients to evaluate their predictive value. ResultsIn 18 eyes in group A and 22 eyes in group B, VZV and HSV was infected in 16 and 2 eyes and 21 and 1 eyes, respectively. The VZV-DNA and HSV-DNA load were compared between the two groups, the difference was statistically significant (Z=－3.762, P＜0.001); compared with group B, the concentrations of VEGF in humor (Z=－3.996), BFGF (Z=－2.430), IL-6 (Z=－3.303), IL-8 (Z=－3.480), and IL-10 (Z=－3.409) increased significantly in group A, the difference was statistically significant (P＜0.05); there was no statistically significant difference in VCAM between the two groups (Z=－0.054, P=0.957). The ROC curve analysis showed that the AUC of VEGF, nucleic acid copies, IL-10, IL-8, IL-6, and bFGF for predicting RD in ARN was 0.871, 0.848, 0.828, 0.823, 0.806, 0.737, respectively. The AUC of combination of VEGF, IL-10, IL-8, IL-6, and BFGF predicted the RD in ARN was 0.924. The optimal cut-off value of nucleic acid copies was 0.40×106, and IL-10, IL-8 and IL-6 were 50.65, 1 695.50 and 6 634.0 pg/ml, respectively. ConclusionAqueous humor viral load as well as cytokines alone or in combination have a reference value for predicting RD secondary to ARN.
Objective To detect the levels of vascular endoth elial growth factor (VEGF) in aqueous humor and vitreous of patients with neovascular glaucoma (NVG) and infer their possible effect on the development of neovascularization of iris. Methods The concentration of VEGF in 22 samples of ocular fluid of aqueous humor and vitreous respectively obtained from 11 patients with NVG undergone intraocular surgery were measured by using enzyme linked immunosobent assay (ELISA) for quantitative analysis. As control, 12 samples of ocular fluid of 6 patients with macular hole were detected by the same methods. Results The mean [AKx-]plusmn;s VEGF concentrations in aqueous humor and vitreous from patients with NVG were [(1.451plusmn;0.247)、(1.610plusmn;0.125) ng/ml] higher than those in the cotrol group [(0.189plusmn;0.038)、(0.201plusmn;0.055) ng/ml], there was a significant difference between the two groups statistically (t=12.007，Plt;0001；t=26.0 57，Plt;0.001). Conclusion The patients with NVG have significantly increased level of VEGF in ocular fluid, and VEGF might fill the role in mediating active iris neovascularization. (Chin J Ocul Fundus Dis, 2001,17:305-306)
ObjectiveTo study the distribution of pathogenic microorganisms in the ocular fluid of patients with acquired immunodeficiency syndrome (AIDS) and infectious uveitis.MethodsIt was a retrospective case analysis. From June 2018 to December 2019, 31 AIDS patients with infectious uveitis who were hospitalized or outpatient at Shanghai Public Health Clinical Center were included in the study. Among them, there were 30 males and 1 female; the average age was 38.51±11.17 years. There were 20 cases of panuveitis, 10 cases of posterior uveitis, and 1 case of infectious endophthalmitis. Serum CD4+T lymphocyte count (CD4+TC) were 0 - 239/μl during the same period. The second-generation gene sequencing technology was used to detect the collected intraocular fluid. Among 31 specimens, aqueous humor and vitreous humor were 27 and 4 respectively.ResultsAmong 31 specimens, 18 samples (58.1%, 18/31) of cytomegalovirus (CMV) were detected; varicella-zoster virus (VZV) were detected in 5 samples (16.1%, 5/31); Epstein-Barr virus were detected in 9 samples (29.0%, 9/31); human beta herpes virus type 6 (HHV6) were detected in 3 samples (9.7%, 3/31), human papillary molluscum virus (HPV), human polyoma virus, type G hepatitis virus were separately detected in 1 sample (3.2%, 1/31), all coexisting with other microorganisms. Parvovirus were detedcted in 8 samples (25.8%, 8/31); treponema pallidum were detedcted in 5 samples (16.1%, 5/31); toxoplasma gondii and Harmon coccidia were detedcted in 1 sample (3.2%, 1/31); synitelium Polycarpum were detedcted in 1 sample (3.2%, 1/31); mycobacterium tuberculosis complex, fungi, and microbacteria coexist were detedcted in 1 sample (3.2%, 1/31). Among the 18 CMV specimens, the number of gene sequences was more than 1059 (50.0%), and 104-1055 (27.7%). Among the 5 specimens of VZV, the number of gene sequences was＞1044 (80.0%). In one specimen, the mycobacterium tuberculosis complex, fungi, and microbacteria coexist, and the number of gene sequences were all＜100. The number of gene sequences of HHV6, HPV, human polyoma virus, type G virus, and parvovirus in all specimens was small. Among 31 specimens, 15 (48.4%) of pathogenic microorganisms were detected at least 2 species.ConclusionsCMV and VZV are the main pathogenic microorganisms of infective uveitis in patients with serum CD4+TC＜100/μl; treponema pallidum, toxoplasma gondii or other protozoa, mycobacterium tuberculosis, and fungi cause more infectious uveitis which are common in AIDS patients with serum CD4+TC＞100/μl. The coexistence of two or more microorganisms can be detected in the intraocular fluid of AIDS patients with infectious uveitis.
ObjectiveTo observe aqueous cytomegalovirus (CMV) DNA load in patients with cytomegalovirus retinitis (CMVR) after allogeneic hematopoietic stem cell transplantation (Allo-HSCT), and to explore influencing factors for transient elevation of CMV-DNA load during the treatment. MethodsA retrospective study. From January 2016 to July 2020, 28 eyes of 19 patients with CMVR after Allo-HSCT diagnosed in the Department of Ophthalmology of Peking University People's Hospital were included in the study. Among them, there were 8 males with 12 eyes, 11 females with 16 eyes; the mean age was 28 years; 10 patients were unilateral and 9 patients were bilateral. During the course of treatment and follow-up, the blood CMV-DNA remained negative. All patients were treated with intravitreal injection of 60 mg/ml ganciclovir 0.05 ml (containing ganciclovir 3 mg), twice a week for two weeks in induction phase and weekly injection in maintenance phase. Aqueous humor sample was collected during injection of ganciclovir (IVG) and CMV-DNA load was determined by real-time quantitative polymerase chain reaction. Intravitreal treatment was terminated if aqueous CMV-DNA load turned negative after the fourth or later intravitreal injection. The patients were followed up every 2 weeks for at least 6 months. Serum CMV-DNA was negative in all patients during treatment and follow-up. All the eyes were divided into continuous decline group and non-continuous decline group depending on whether there was transient elevation of aqueous CMV-DNA load, and data between two groups were compared. Pearson linear regression analysis was used to analyze the correlation between aqueous CMV-DNA load and injection times or treatment duration. ResultsAt the end of treatment, the median number of IVG in the affected eye was 7 (4, 9). The results of correlation analysis showed that the aqueous humor CMV-DNA load of the affected eye was related to the number of treatments [R2=0.385, P＜0.000 1, B=－0.237 log10 copies/(ml · time)], and the duration of treatment [R2=0.394, P <0.000 1, B=－0.301 log10 copies/(ml · week)] were negatively correlated. Among the 28 eyes, 13 eyes (46.4%, 13/28) in the continuous decline group and 15 eyes (53.6%, 15/28) in the non-sustained decline group. Baseline visual acuity (t=－1.223), intraocular pressure (t=1.538), aqueous humor CMV-DNA load (t=－0.109), retinitis lesion area (Z=－0.308) in the continuous decline group and the non-continuous decline group), the number of quadrants involved (Z=－0.024) and whether the macula was involved (Z=－1.826), combined with anterior segment inflammation (Z =－0.499), combined with high intraocular pressure (Z=－1.342), terminal visual acuity (t =－0.845), intraocular pressure (t=－0.068), total IVG times (Z=0.907), age (Z=－0.832), gender composition (Z=－1.074), etc. The difference was not statistically significant (P＞0.05). ConclusionThe CMV-DNA load in aqueous humor decreases by about 50% every week during the treatment of CMVR eyes after Allo-HSCT; the transient increase in the CMV-DNA load in the aqueous humor during treatment does not affect the treatment process and clinical prognosis.
ObjectiveTo compare the clinical manifestations of ocular toxoplasmosis (OT) in adult and children, and to preliminarily explore the role of intraocular fluid detection in the early diagnosis of OT.MethodsA retrospective study. From January 2018 to October 2019, 60 cases of OT patients with 60 eyes diagnosed in the Department of Ophthalmology of Beijing Chaoyang Hospital Affiliated of Capital Medical University were included in the study. The medical history information of patients was collected in parallel with slit-lamp microscopy, indirect ophthalmoscope examination, and canine toxoplasma antibody detection in aqueous or vitreous fluid. Fifty-eight cases underwent visual inspection; 2 cases did not underwent visual inspection, who were children. The visual acuity examination was carried out using the new version of the standard logarithmic visual acuity chart, which was converted into the logarithmic minimum angle of resolution (logMAR) visual acuity during statistics. According to age, the patients were divided into adult group and child group, with 12 eyes in 12 cases and 48 eyes in 48 cases, respectively. The clinical characteristics and main points of diagnosis and treatment of the two groups of patients were compared and observed. The comparison among the measurement data groups conforming and the normal distribution was performed by the independent t test. The comparison between the measurement data groups of the skewed distribution was performed by the Kruskal-Wallis test. The qualitative data were compared with χ2 test.ResultsAmong the adult group and the child group, 7 (58.3%, 7/12) and 34 (70.8%, 34/48) patients with a clear history of contact with dogs and cats were in the adult group and the child group, respectively. The adult group was significantly lower than the child group, however, there was no different statistical significance (χ2=0.236, P=0.627). At the first visit, the self-reported blurred vision of the adult group and the child group was 10 (83.3%, 10/12) and 22 (45.8%, 22/48) cases, respectively. In the adult group and the child group, 3 (25.0%, 3/12) and 20 (43.5%, 20/46) eyes with logMAR visual acuity greater than 1.85, 8 (66.7%, 8/12) and 22 (45.8%, 22/46) eyes with logMAR visual acuity less than 0.3. The visual acuity of the adult group was better than that of the child group, and the difference was statistically significant (Z=2.162, P=0.031). There was no statistically significant difference in the composition ratio of different clinical types of the two groups of eyes (χ2=1.908, P=0.385). The incidence of inflammation in the anterior segment of the eye in the adult group and the child group were 25.0% (3/12) and 56.3% (27/48), respectively; there was no statistically significant difference between the two groups (χ2=3.750, P=0.053). The concentration of antibodies in the vitreous humor of the affected eye in the adult group and the child group was greater than that of aqueous humor. The antibody concentrations of vitreous humor and aqueous humor were 36.51 (22.58) and 19.94 (21.78) U/ml in the children group; 45.95 (56.44) and 32.20 (38.64) U/ml in the adult group. Comparison of antibody concentrations in the vitreous humor and aqueous humor of the affected eyes in the child group showed statistically significant differences (Z=−1.984, P=0.047).ConclusionsCompared with children with OT, adult patients with OT have better vision and mild inflammation or hyperplasia of the vitreous cavity. The detection of antibodies related to toxoplasma in the intraocular fluid is helpful for early diagnosis.
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 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)
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%.