Background Herpes stromal keratitis (HSK) is a common infectious ocular surface disease,with a higher recurrent rate,especially in necrotizing HSK.The diagnosis of HSK primarily depends on signs and symptoms,and specific laboratory diagnostic is lack.Objective This study was to clarify the expression of herpes simplex virus (HSV) in the corneal epithelium scrapings and tears of necrotizing HSK patients.Methods Thirty eyes of 30 patients with necrotizing HSK were enrolled in Nanjing Hospital of Nanjing Medical Hospital from September 2012 to September 2013 under the patient's informed consent.The eyes were examined by slit lamp microscope and scored.HSK patients received local and systemic therapy for 8 weeks and then an oral maintenance dose for 6 months.Corneal epithelial scrapings and tears samples were collected for HSV DNA detection by real-time PCR before and the 1 st,2nd,4th,6th and 8th week after therapy respectively.The difference of HSV positive rate was compared between corneal epithelium scrapings and tears samples using Chi-square test.Multilevel mixed effective model was employed to evaluate HSV concentration change in the samples at various time points in the HSV-positive patients of initial visit.The correlation between HSV concentration and clinical score was analyzed by Spearman rank correlation.Results HSV-positive rate was 46.4％ (13/30) in the corneal epithelial scrapings and 13.3％(4/30) in the tear samples,showing a significant difference between them (P =0.006).HSV-positive rate was significantly lower in the corneal epithelial scrapings 1 week,2 weeks and 4 weeks after treatment than before (P =0.001,0.003,0.004),and no HSV was detected 6 weeks and 8 weeks after treatment.No significant change in HSV-positive rate in tear samples in 1 week and 2 weeks after treatment in comparison with before treatment (P =1.000,0.583),and no HSV was detected after 4 weeks following treatment.The HSV concentration was 2460 (2 165-636500)/ml in initial 13 HSV-positive eyes of corneal epithelial scrapings and 0 (0-1150)/ml in initial 13 HSV-positive eyes of tear samples.Multilevel mixed effective model determined that HSV concentration was significantly lower in corneal epithelial scraping than that in tear (P =0.005),and HSV concentration was reduced with the lapse of time (P =0.001),with the faster rate of decline in the corneal epithelial scrapings (P =0.049).A positive correlation was found between initial HSV concentration and clinical scores (rs =0.844,P =0.000).Conclusiors Real-time PCR appears to be a powerful molecular tool for the detection of HSV in the HSK,especially in corneal epithelial scrapings of lesion.The initial positive outcome of viral DNA in corneal epithelial scrapings predicts a severe clinical procedure.

OBJECTIVETo evaluate the effectiveness of small interfering RNA (siRNA) on inhibiting severe acute respiratory syndrome (SARS)-associated coronavirus replication, and to lay bases for the future clinical application of siRNA for the treatment of viral infectious diseases.

METHODSVero-E6 cells was transfected with siRNA before SARS virus infection, and the effectiveness of siRNA interference was evaluated by observing the cytopathic effect (CPE) on Vero-E6 cells.

RESULTSFive pairs of siRNA showed ability to reduce CPE dose dependently, and two of them had the best effect.

CONCLUSIONsiRNA may be effective in inhibiting SARS-associated coronavirus replication.

Animals ; Cercopithecus aethiops ; RNA, Small Interfering ; pharmacology ; SARS Virus ; drug effects ; Transfection ; Vero Cells ; Virus Replication ; drug effects

Reynoutria japonica Houtt., belonging to Polygoneae of Polygonaceae, is a Chinese medicinal herb with the functions of draining dampness and relieving jaundice, clearing heat and detoxifying, dispersing blood stasis and relieving pain, and relieving cough and resolving phlegm. In this study, we carried out high-throughput sequencing for the chloroplast genome sequences of five cultivars of R. japonica and analyzed the genome structure and variations. The chloroplast genomes of the five R. japonica cultivars had two sizes (163 376 bp and 163 371 bp) and a typical circular tetrad structure composed of a large single-copy (LSC) region of 85 784 bp, a small single-copy (SSC) region of 18 616 bp, and a pair of inverted repeat (IR) regions (IRa/IRb) which are spaced apart. A total of 161 genes were obtained by annotation, which consisted of 106 protein-coding genes, 10 rRNA-coding genes, and 45 tRNA-coding genes. The total GC content was 36.7%. Specifically, the GC content in the LSC, SSC, and IR regions were 34.8%, 30.7%, and 42.7%, respectively. Comparison of the whole chloroplast genome among the five cultivars showed that trnk-UUU, rpoC1, petD, rpl16, ndhA, and rpl12 in coding regions had sequence variations. In the phylogenetic tree constructed for the 11 samples of Polygoneae, the five cultivars of R. japonica clustered into one clade near the root and was a sister group of Fallopia multiflora (Thunb.).
Base Composition ; Genome, Chloroplast/genetics* ; Open Reading Frames ; Phylogeny ; Reynoutria