There are many types of fundus diseases and their causes are complex. They can be caused by metabolic factors or inflammatory factors. Fundus examination and imaging examination tools are the main methods for diagnosing fundus diseases. However, in terms of determining the cause and early diagnosis, if the intraocular fluid detection technology can be reasonably combined, the advantages will be greater. Intraocular fluid is the general term for fluid in the eyeball, including aqueous humor, vitreous humor, etc. The molecular components that can be tested include DNA, RNA, antigens, antibodies, and cytokines. With the advancement of molecular testing technology and equipment, intraocular fluid testing as an evidence-based method has gradually been incorporated into the consensus and guidelines of more fundus disease experts, and is mainly used for infectious fundus diseases and camouflage syndromes. Reasonable use of intraocular fluid testing can help improve the personalized diagnosis and treatment of fundus diseases and reduce unnecessary drug overuse. However, it is worth noting that intraocular fluid detection is only one of many tools and cannot replace other examinations and clinical experience. Excessive intraocular fluid testing not only increases the risk of clinical infections because of invasiveness, but also increases the burden on patients.
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 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 study the changes and correlation of cytokines in aqueous humor before and after intravitreal injection of conbercept (IVC) treatment in patients with proliferative diabetic retinopathy (PDR).MethodsA prospective clinical study. From March to December 2019, 36 patients (42 eyes) of PDR patients treated with IVC combined with pars plana vitrectomy (PPV) (the observation group) and 27 patients (31 eyes) underwent cataract surgery in the same period (control group) in Department of Ophthalmology of the First Affiliated Hospital of Guangzhou University of Chinese Medicine were included in this study. Before PPV 5-7 days, IVC treatment was performed, and the aqueous humor were extracted during IVC and second-stage PPV in the observation group. The aqueous humor was extracted during cataract surgery in the control group. Luminex assay was used to detect VEGF-A, placental growth factor (PLGF), platelet-derived growth factor-AA (PDGF-AA), platelet-derived growth factor-BB (PDGF-BB), angiopoietin-like protein 4 (ANGPTL4), IL-6, IL-8, IL-1β, monocyte chemoattractant protein-1 (MCP-1), intercellular adhesion molecule-1 (ICAM-1), tumor necrosis factor-α (TNF-α) cytokine expression. For normally distributed data, the independent sample t test was used for comparison between two independent samples; for non-normally distributed data, the Wilcoxon rank sum test was used for comparison between two independent samples. The correlation analysis used Spearman rank correlation test.ResultsBefore IVC treatment, the concentrations of VEGF-A, PLGF, PDGF-AA, ANGPTL4, IL-6, IL-8, MCP-1 and ICAM-1 in the aqueous humor of PDR patients were significantly higher than those in the control group (P＜0.05). After IVC treatment, the concentration of VEGF-A in the aqueous humor was significantly lower than that before treatment, and the concentrations of ANGPTL4 and IL-8 were significantly higher than those before treatment (P＜0.05). There were no significant differences in the concentrations of PLGF, PDGF-AA, PDGF-BB, IL-6, IL-1β, MCP-1, ICAM-1 and TNF-α before and after IVC treatment (P＞0.05). Before IVC treatment, the concentration of VEGF-A was positively correlated with PLGF, PDGF-AA, PDGF-BB, ANGPTL4, IL-6, IL-8, MCP-1 and TNF-α (P＜0.05).ConclusionsIVC treatment can reduce the concentration of VEGF-A and increase the concentrations of ANGPTL4 and IL-8 in aqueous humor in PDR patients before PPV.
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.
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.
ObjectiveTo observe the safety and efficacy of regime that based on aqueous cytomegalovirus-DNA (CMV-DNA) load and IL-8 determination for therapeutic monitoring and local treatment cessation of cytomegalovirus retinitis (CMVR) patients after allogeneic hematopoietic stem cell transplantation (HSCT).MethodsA prospective case series study. A total of 14 CMVR patients (22 eyes) after allogeneic HSCT diagnosed in Ophthalmology Department of Peking University People's Hospital between January 2016 and December 2018 were involved in this study. All patients were CMV-DNA seronegative at baseline and were treated with intravitreous injection of ganciclovir (IVG, 3 mg in 0.05 ml) twice per week for 4 times in the induction stage and once a week in the maintenance stage. Aqueous humor sample was collected during the first time of IVG every week. CMV-DNA and the level of IL-8 were measured by real time quantitative PCR and ELISA, respectively. During follow-up, negative CMV-DNA (＜103/ml) or level of IL-8＜30 pg/ml in aqueous sample was set as local treatment cessation. Then patients were followed every 2 weeks for at least 6 months. BCVA, intraocular pressure and fundus examination were taken for each visit. The BCVA examination was performed using the international standard visual acuity chart, which was converted into logMAR visual acuity. BCVA and intraocular pressure at the baseline and the last follow-up were compared by the Student t matching test.ResultsOf the 14 CMVR patients (22 eyes) after allogeneic HSCT, 8 patients (16 eyes) were bilateral, 6 patients (6 eyes) were unilateral. At the baseline, the mean logMAR BCVA was 0.814±0.563, the intraocular pressure was 17.2±7.8 mmHg (1 mmHg=0.133 kPa), the mean aqueous CMV-DNA load was (3.43±4.96)×105/ml, the mean level of IL-8 was 518±541 pg/ml. At cessation of local treatment, the median number of intravitreal injections was 5 times. Nine eyes showed negative CMV-DNA in aqueous humor, of which, 7 eyes showed negative IL-8 in aqueous. CMV-DNA could still be detected in 13 eyes, while IL-8 was negative. Only one eye’s retinal lesion was completely quiet. Six months after local treatment cessation, the mean logMAR BCVA was 0.812±0.691, the intraocular pressure was 14.8±5.4 mmHg; which was not significantly different from baseline (t=-0.107, 1.517; P=0.916, 0.137). Recurrence of CMVR happened in only 1 eye because of systemic EB virus infection. Retinal lesions progressively improved and became completely quiet in all the remaining 20 eyes. In 22 eyes, iatrogenic vitreous hemorrhage occurred due to low platelet count during treatment (＜30×109/ml) in 4 eyes. When the treatment was terminated for 6 months, the fundus of hematoma absorption was clearly visible. At the time of CMVR diagnosis, there were 2 eyes (9%) with posterior subcapsular opacity, which may be caused by systemic glucocorticoid therapy after allogeneic HSCT.ConclusionAqueous CMV-DNA load and level of IL-8 could be used as quantitative variables for monitoring the therapeutic effect and determining time for local treatment cessation for CMVR after HSCT safely and efficiently.
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)
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.
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)
