ObjectiveTo detect the effect of adeno-associated-virus induced Kringles5 gene on retinal neovascularization in rats with retinopathy of prematurity (ROP), and to explore the new ways of treatment for ROP.MethodspSNAV-Kringle5-gfp carrier was constructed by subclone and adeno-associated-virus was packed to form rAAV-Kringle5-gfp. ROP model was set up under circumstances of high oxygen in 21 SD rats which were divided into experimental (21 eyes) and control group (21 eyes). Eighteen eyes from each group was used to making the histologic section of retina, and the other 3 eyes in each group was detected by polymerase chain reaction (PCR) and Western blotting. There were 5 rats in the normal control group. AAV-Kringle5-gfp with the dosage of 10 μl and titer of 2.5×1012vg/ml was injected into the eyes in experimental group, while rAAVlacZ with the same dosage and titer of 2.5×1011vg/ml was injected in to the eyes in control group. The expression of target gene in ocular tissues was observed under the fluoroscope. Twelve weeks later, the rats were executed, and the staining of Ⅷ factor related antigens in retinal vascular endothelial cells was performed and number of nucleolus of vascular endothelial cells were counted. ResultsThe plasmid of pSNAV-Kringle5-gfp was correct according to the sequence measurement; the expression of rAAV-Kringle5-gfp was found in vitreous cavity and on retina; the expression of target gene was found on the level of mRNA and protein; the number of nucleolus of vascular endothelial cells on the surface of retina was (19.954 2±3.825 7) in experimental group and (7.335 2±2.731 3) in the control group, which had significant difference between the two groups (P＜0.01）.ConclusionsAdeno-associated-virus induced Kringles5 gene can inhibit the occurrence of retinal neovascularization in patients with ROP.(Chin J Ocul Fundus Dis, 2005,21:288-291)
【摘要】 目的 评价高效抗逆转录病毒疗法（highly active antiretroviral therapy，HAART）对艾滋病患者的疗效，并观察其耐药情况。 方法 2008年11月，对23例接受免费HAART治疗1~3年的艾滋病患者的HIV-RNA病毒载量、HIV-1耐药、CD4+T淋巴细胞绝对计数、常规生化检测结果进行分析。 结果 23例艾滋病患者在HAART治疗1~3年后有6例（26.1%）患者HIV-RNA载量在水平线以下，17例（73.9%）HIV-RNA载量未达到检测线以下。5例患者（21.7%）的HIV-1发生了耐药，服药依从性良好的患者仅9例（39.1%）。 结论 HAART对艾滋病患者的疗效较差、耐药率高、服药依从性差。【Abstract】 Objective To evaluate the efficacy of highly active antiretroviral therapy (HAART) in treating HIV, and observe the resistance of HAART in AIDS patients. Methods Twenty-three HIV patients receiving free HAART for one to three years were investigated. The plasma viral load, drug resistance to HIV-1, CD4+ T cell count were tested and routine laboratory examinations were performed in our study. Results After one to fhnee-year HAART treatment, HIV viral load of six patients (26.1%) declined to the undetectable level, while the viral load of 17 patients (73.9%) kept at a high level. Resistance to HIV-1 occurred in five cases (21.7%). Only nine patients (39.1%) had good treatment compliance. Conclusion HAART has a poor therapeutic effect on patients with HIV/AIDS with a high rate of drug resistance and poor treatment compliance.
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 observe the changes of varicella zoster virus (VZV)-DNA load in aqueous humour samples in VZV-induced acute retinal necrosis (ARN) in the early stages of antiviral treatment. MethodsA retrospective observational clinical study. From April 2016 to April 2018, 24 patients with 24 eyes of VZV-induced ARN who were diagnosed by Department of Ophthalmology, Eye and ENT Hospital of Fudan University and received complete aqueous humor sampling were included in the study. Among them, there were 13 males with 13 eyes, 11 females with 11 eyes; 12 left eyes and 12 right eyes; the age was 52.0±9.5 years old (39-71 years old). The time from the onset of ocular symptoms to the diagnosis of ARN was 16.6±6.1 days (7-30 days). Best-corrected visual acuity (BCVA) and ultra-wide-field fundus imaging were performed in all affected eyes. The BCVA examination was carried out using the Snellen visual acuity chart, which was converted into the logarithm of the minimum angle of resolution (logMAR) visual acuity. All patients were given intravitreal injection of 40 mg/ml ganciclovir 0.1 ml (including 4 mg of ganciclovir), 2 times a week, until the active necrotizing retinal lesions subsided, at most after the diagnosis 4 weeks, with a maximum of 9 injections. The follow-up period was 12.8±5.6 months. The aqueous humor samples were collected at presentation and 4, 7, 14, 21, 28 days after the initiation of antiviral therapy, and the VZV-DNA load was detected by real-time quantitative polymerase chain reaction. A plateau phase and a logarithmic reduction phase of the DNA load changes were observed after antiviral treatment began. Wilcoxon rank sum test was used to compare and analyze the differences in BCVA between the eyes at baseline and last follow-up. ResultsThe mean viral load at presentation was 8.6×107±1.3×108 copies/ml. The initial plateau phase last for an average of 7.4±2.4 days. In the following logarithmic reduction phase, the mean slope of the decline in viral load was －0.13±0.04 log/day, and the expected time for half reduction of the initial viral load was 2.5±0.7 days. After 28 days antiviral treatment, the viral load decreased to 1.7×105±1.8×105 copies/ml. In the course of the disease, rhegmatogenous retinal detachment occurred in 16 eyes. Before treatment and at the last follow-up, the logMAR BCVA of the affected eye was 1.1±0.6 and 0.8±0.7, respectively. The results of correlation analysis showed that the logMAR BCVA at the last follow-up was correlated with the initial VZV-DNA load (r=0.467, P=0.033). ConclusionThe VZV-DNA load in the aqueous humor of eyes with VZV-induced ARN is significantly decreased after antiviral treatment, which is closely related to the clinical process of ARN.