1.Changes of viral DNA load in the early stages of antiviral treatment in varicella zoster virus-induced acute retinal necrosis
Boya LEI ; Zhujian WANG ; Ruiping GU ; Qinmeng SHU ; Rui JIANG ; Qing CHANG ; Gezhi XU ; Min ZHOU
Chinese Journal of Ocular Fundus Diseases 2022;38(5):371-375
Objective:To 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.Methods:A 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.Results:The mean viral load at presentation was 8.6×10 7±1.3×10 8 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 1.7×10 5±1.8×10 5 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). Conclusion:The 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.
2.Effect of Acupuncture at “Zhibian (BL 54)-to-Shuidao (ST 28)” on Key Factors of Endogenous Exogenous Apoptotic Pathways in Asthenospermia Model Mice
Jianheng HAO ; Jia REN ; Boya CHANG ; Haijun WANG ; Laixi JI
Journal of Traditional Chinese Medicine 2024;65(1):86-93
ObjectiveTo observe the effect of acupuncture “Zhibian (BL 54)-to-Shuidao (ST 28)” on the reproductive function of asthenospermia model mice and to explore the possible mechanism. MethodsForty-two male C57BL/6 mice were randomly divided into blank group, model group, and acupuncture group, with 14 mice in each group. Cyclophosphamide 30 mg/(kg·d) was injected intraperitoneally for 7 days to establish model of asthenospermia for the model group and the acupuncture group, while 0.9% sodium chloride solution 10 ml/(kg·d) was injected intraperitoneally for 7 days in the blank group. After successful modeling, mice in the acupuncture group received acupuncture at “Zhibian (BL 54)-to-Shuidao (ST 28)” once a day for 20 minutes, 6 times a week for 3 weeks; the remaining two groups were fixed without acupuncture. Daily observations were made on the general conditions of mice in all groups, and changes in body weight after intervention were recorded. On the next day after completing the treatment, 6 male mice selected randomly from each group to test the sperm quality as well as testicular and epididymal weights, and calculate the corresponding indices; ELISA was used to test the levels of testosterone (T), follicle-stimulating hormone (FSH), and luteinizing hormone (LH) in serum; HE staining and TUNEL staining were performed to observe the pathological morphology and apoptosis of testicular and epididymal tissues; fluorescence quantitative PCR and western blot were used to detect changes in the expression of apoptosis-related factors (Fas), Fas-associated death domain protein (FADD), B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and cysteine-aspartic acid protease (caspase)-3 mRNA and protein in testicular tissue. The remaining 8 male mice in each group were housed with estrus-cycling mice of the same strain at 1∶1 ratio, and the pregnancy rate and number of embryos per litter in each group were determined after mating. ResultsIn comparison with the blank group, mice in the model group exhibited reduced body weight, decreased testicular mass, testicular index, epididymal mass, and epididymal index. Additionally, there was a decrease in total sperm count, sperm motility, and sperm viability. Serum levels of T, FSH, and LH were reduced. The apoptosis rate increased, and the expression levels of Fas, FADD, Bax, and Caspase-3 mRNA and proteins were elevated, while Bcl-2 mRNA and protein expression levels decreased (P<0.01). Pathological abnormalities were observed in testicular and epididymal tissues, with disrupted arrangement of seminiferous tubules and a decreased number of spermatogenic cells within the tubular lumen. Furthermore, the pregnancy rate and the number of embryos per litter decreased significantly after mating (P<0.01). In comparison with the model group, mice in the acupuncture group showed improvements in testicular mass, testicular index, epididymal mass, epididymal index, total sperm count, sperm motility, and sperm viability. Serum levels of T, FSH, and LH increased. The apoptosis rate decreased, and the expression levels of Fas, FADD, Bax, and Caspase-3 mRNA and proteins decreased, while Bcl-2 mRNA and protein expression levels increased (P<0.05 or P<0.01). Morphological improvements were observed in testicular and epididymal tissues, with a compact arrangement of seminiferous tubules and an increased number of spermatogenic cells within the tubular lumen. Furthermore, the pregnancy rate and the number of embryos per litter increased significantly after mating (P<0.01). ConclusionAcupuncture “Zhibian (BL 54)-to-Shuidao (ST 28)” can improve testicular tissue apoptosis and enhance reproductive function in a mouse model of asthenospermia. Its mechanism may be associated with the modulation of key factors in the extrinsic membrane receptor pathway (Fas-mediated pathway) and the intrinsic mitochondrial pathway (Bcl-2/Bax-regulated pathway) in testicular tissue.