ABSTRACT
AIM@#To evaluate the anti-HIV activity and mechanism of action of wikstroelide M, a daphnane diterpene from Daphne acutiloba Rehder (Thymelaeaceae).@*METHODS@#The anti-HIV activities of wikstroelide M against different HIV strains were evaluated by cytopathic effect assay and p24 quantification assay with ELISA. The inhibitory effect of wikstroelide M on HIV reverse transcription was analyzed by real-time PCR and ELISA. The effect of wikstroelide M on HIV-1 integrase nuclear translocation was observed with a cell-based imaging assay. The effect of wikstroelide M on LEDGF/p75-IN interaction was assayed by molecular docking.@*RESULTS@#Wikstroelide M potently inhibited different HIV-1 strains, including HIV-1IIIB, HIV-1A17, and HIV-19495, induced a cytopathic effect, with EC50 values ranging from 3.81 to 15.65 ng·mL⁻¹. Wikstroelide M also had high inhibitory activities against HIV-2ROD and HIV-2CBL-20-induced cytopathic effects with EC50 values of 18.88 and 31.90 ng·mL⁻¹. The inhibitory activities of wikstroelide M on the three HIV-1 strains were further confirmed by p24 quantification assay, with EC50 values ranging from 15.16 to 35.57 ng·mL⁻¹. Wikstroelide M also potently inhibited HIV-1IIIB induced cytolysis in MT-4 cells, with an EC50 value of 9.60 ng·mL⁻¹. The mechanistic assay showed that wikstroelide M targeted HIV-1 reverse transcriptase and nuclear translocation of integrase through disrupting the interaction between integrase and LEDGF/p75.@*CONCLUSION@#Wikstroelide M may be a potent HIV-1 and HIV-2 inhibitor, the mechanisms of action may include inhibition of reverse trascriptase activity and inhibition of integrase nuclear translocation through disrupting the interaction between integrase and LEDGF/p75.
Subject(s)
Humans , Anti-HIV Agents , Pharmacology , Therapeutic Uses , Cell Line , Daphne , Chemistry , Diterpenes , Pharmacology , HIV Infections , Drug Therapy , Virology , HIV Integrase , Metabolism , HIV Integrase Inhibitors , Pharmacology , Therapeutic Uses , HIV Reverse Transcriptase , HIV-1 , HIV-2 , Intercellular Signaling Peptides and Proteins , Metabolism , Phytotherapy , Plant Extracts , Pharmacology , Therapeutic Uses , Virus Integration , Virus ReplicationABSTRACT
Objective:To investigate the role of human telomerase reverse transcriptase(hTERT) in carcinogenesis of oral submucous fibrosis(OSF).Methods:The expression of hTERT was examined by immunohistochemical SP method in 10 cases of normal oral mucosa, 46 cases of OSF,14 of carcinogenesis of OSF and 10 of adjacent non-cancerous OSF tissue. Results:The expression of hTERT was negative in normal oral muscosal epithelium.The frequency of hTERT positive expression was higher in carcinogenesis lesions of OSF(12/14) than those in OSF(7/46)(P
ABSTRACT
Objective DNA replication increases during tumor cells limitlessly proliferate. To develop a method that can early diagnose and routine screen out bladder cancer, but it does not impair patients by detecting cdc6 in exfoliate cell in urine of patients with bladder cancer. Methods 200 ml fresh urine of patients with bladder cancer or non bladder cancer who voided urine after morning was collected and centrifugated for urine sediment, then total RNA was extracted with TRIZOL reagent from it and detected the expression of cdc6 gene by RT PCR. Results 28 samples were positive and 2 samples were negative in 30samples of bladder cancer, positive rate was 93.3%. Positive rate was 16.7% in 30 samples of non bladder cancer. Compared with the sensitivity of urine cytology, that of cdc6 RT PCR increases markedly ( P