Objective To amplificate,clone and sequence the thymidine kinase (TK) gene of herpes simplex virusⅡ(HSVⅡ); to construct and appraise the fusion gene eukaryon expression vector, pcDNA3/HSVⅡ TK/angiostatin. MethodsThe Hep2 cells were infected by HSVⅡ Sav strain. HSVⅡ genomic DNA was purified from the Hep2 cells suspension and used as template to run PCR for TK gene amplification. The amplified products were cloned into PC DNA3 vector and sequenced. The vector pcDNA/HSVⅡ TK was cut by endonuclease. The gained TK gene was cloned into eukaryon expression vector. pcDNA3/angiostation, which had been constructed. ResultsCoding region of HSVⅡTK gene consisted of 1 128 bp except stop code, it encoded 376 amino acids.After cutting the new vector by endonuclease Hind Ⅲ and BamH Ⅰ,we gained the following gene fragment: 1000 bp (TK) and 700 bp (angiostation).Conclusion The fusion gene eukaryon expression vector， pcDNA3/HSVⅡ TK/angiostatin has been constructed.
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.
ObjectiveTo explore the pathogenic mechanism of human herpesvirus 6B (HHV-6B) in the intractable mesial temporal lobe epilepsy (MTLE).Methods42 cases of intractable MTLE and 17 cases of non-MTLE patients who treated in Wuhan Brain Hospital from Jan. 2012 to Dec. 2018 were enrolled in this study. Resected hippocampus, amygdala and mixed uncus samples were examined by real-time polymerase chain reaction (PCR) and reverse-transcriptase PCR to detect viral DNA and message RNA. Comparative analysis was used between the clinical characteristics and the HHV-6 detection.ResultsDetection of HHV-6 DNA was higher in MTLE patients (40.48%) than non-MTLE patients (11.7%). HHV-6B viral DNA was determined in 15/30 HHV-6 DNA-positive samples, and no HHV-6B mRNA were detected in all HHV-6 DNA negative samples. The patient’s age was the only influencing factor of HHV-6 expressing in MTLE. While the patient’s gender, onset age and possible disease cause had no effect on the HHV-6 content.ConclusionsThis study suggests that HHV-6B may play an important role in the pathogenesis of MTLE, but the mechanism needs further study.
Objective To investigate the general state, psychologic status and health related quality of life (HRQOL) in patients with post-herpetic neuralgia (PHN), and explore the influencing factors on HRQOL and their major and minor relationship. Methods From August 2014 to August 2015, a questionnaire survey was conducted to patients with PHN. The data of the patients’ general situation was colleted, the psychologic status was assessed by Self-rating Depression Scale (SDS), and the HRQOL of the patients was assessed by 36-item Short Form Survey. One-way ANOVA and stepwise regression analysis were used to analyze the influencing factors of HRQOL. Results A total of 177 questionnaires were issued, and 158 were recovered with a response rate of 89.3%. The average score of HRQOL of the 158 patients was 513.67±119.79, and the average score of SDS was 43.65±11.34. SDS scores showed no depression in 41 patients (25.9%), mild depression in 47 (29.7%), moderate depression in 39 (24.8%), and severe depression in 31 (19.6%). Whether receiving regular treatment, course of PHN, SDS score, physical exercise, sleep time and age were the main influencing factors of HRQOL. Conclusions Most patients with PHN suffer from depressive symptoms with poor HRQOL scores. Receiving early treatment with integrated traditional Chinese and western medicine, strengthening exercise, ensuring adequate sleep and receiving psychological counseling can significantly improve the HRQOL of PHN patients.
ObjectiveTo compare the analgesic effect of different treatment for herpes zoster neuralgia, and optimize herpetic neuralgia treatment. MethodsWe collected hospital inpatient data with herpes zoster by the First Military Medical Case Management System between January 1st, 2009 and December 20th, 2013. All the patients were divided into five groups according to different treatments. Patients in group A accepted valaciclovir capsules and doxepin hydrochloride tablets; patients in group B accepted valaciclovir capsules; patients in group C accepted valaciclovir capsule, nefopam hydrochloride tablets and doxepin hydrochloride tablets; patients in group D accepted valaciclovir capsules and nefopam hydrochloride tablets; patients in group E accepted valaciclovir capsules and non-steroidal anti-inflammatory drugs (including nimesulide capsules and ibuprofen sustained-release capsules). We collected such statistic data as sex, age, time of onset, time of pain disappearing after treatment. Then, we analyzed the difference among the groups on the time of pain disappearing. ResultsThere were 336 cases compliant with the standard in all the 898 cases of medical record data. The number of the patients was 72, 86, 66, 60 and 52; while the pain disappearing time after treatment was (5.94±2.54), (8.60±3.09), (5.77±1.85), (5.80±1.96) and (6.86±2.18) days, respectively in group A, B, C, D and E. Pain disappearing time after treatment of group B was significantly different from groups A, C, D, and E (P<0.05); group E was significantly different from groups A, C, and D (P<0.05); there was no significant difference between group A and groups C and D, and between group C and D (P>0.05). ConclusionThe combination of nefopam hydrochloride tablets, doxepin hydrochloride tablets, nimesulide capsules, and ibuprofen sustained-release capsules is effective in the treatment of herpes zoster neuralgia. The use of nefopam hydrochloride tablets with doxepin hydrochloride tablets is more effective than the combination of nimesulide capsules and ibuprofen sustained-release capsules. Doxepin hydrochloride tablets are not significantly different from nefopam hydrochloride tablets. Nefopam hydrochloride tablets and doxepin hydrochloride tablets are not associated with increased efficacy.