Hepacivirus ; physiology ; Hepatitis C ; genetics ; metabolism ; virology ; Host-Pathogen Interactions ; Humans

·Corneal neovascularization ( CNV) is one of the most important causes that affecting corneal transparency, and it is a high risk factor of allogeneic corneal graft rejection. lt has become a research focus for the regulation of CNV. microRNAs are a class of endogenous non-protein-coding micromolecule RNAs which play a critical role in regulating a series of life process.Researches in recent years show a close correlation between microRNA and CNV.ln this article we reviewed the recent advances in these researches.

Drug therapy is one of the efficient methods for prostate cancer treatment. However, drug resistance greatly hindered the treatment of prostate cancer patients. Herein, the mechanisms of drug resistance in prostate cancer have been exhaustively reviewed, and that can provide an alternative strategy and new targets for anti-prostate cancer therapy.
Drug Resistance, Neoplasm ; Humans ; Male ; Prostatic Neoplasms ; drug therapy

OBJECTIVE:To prepare Zuojin gastric-mucoadhesive tablets and evaluate their drug release properties in vitro. METHODS:Zuojin gastric-mucoadhesive tablets were prepared with hydroxypropyl methyl cellulose K15M(HPMC-K15M),car-bomer 934P and HPMC-E50 as the bioadhesive and matrix materials,and basic magnesium carbonate(foaming material),95% al-cohol solution(adhesive)and aerosil(glidant and lubricant)as the adjuvants. With the accumulative release of total alkaloids from Coptis chinensis Franch. at 2,6 and 10 h(Q2 h,Q6 h and Q10 h)as the indexes,orthogonal design test was conducted to optimize the amounts of HPMC-K15M,carbomer 934P,HPMC-E50 and basic magnesium carbonate,and verification was carried out. Drug re-lease properties in vitro of the preparation and Zuojin conventional tablets were observed and in vitro adhesion thereof determined. RESULTS:The optimal formulation was as follows as that for 50 tablets,HPMC-K15M of 0.7 g,carbomer 934P of 0.2 g, HPMC-E50 of 3.5 g and basic magnesium carbonate of 0.4 g. The Q2 h,Q6 h and Q10 h of three batches of prepared samples were 24.32%,56.10% and 77.04% respectively. 1-12 h drug release in vitro of prepared samples was in conformity with Ritger-Peppas equation. The Q2 h of Zuojin conventional tablets and Zuojin gastric-mucoadhesive tablets were 80.46% and 24.04%,Q12 h thereof 92.15% and 95.83% and gastric adhesion thereof 24.2 and 74.0 g/cm2,respectively. CONCLUSIONS:Zuojin gastric-mucoadhesive tablets which have sustained-release effect and adhesive property have been prepared successfully.

Drug therapy is one of the efficient methods for prostate cancer treatment. However, drug resistance greatly hindered the treatment of prostate cancer patients. Herein, the mechanisms of drug resistance in prostate cancer have been exhaustively reviewed, and that can provide an alternative strategy and new targets for anti-prostate cancer therapy.

Objective To explore the effect of transcription factor E2F1 on the apoptosis of small cell cancer line H446. Methods Plamid vector- mediated E2F1 small hairpin RNA (shRNA) was used to silence E2F1 in H446 cell. RT-PCR and western-blot assay were used to detect the expressions of E2F1 and Bcl-2. The apoptosis rate in H446 cell line was detected by flow cytometry assay. Result E2F1 protein was suppressed in shRNA1-modified H446 cell. Sgnificant difference of the apoptosis was shown between E2F1 shRNA1 group and the other two groups. Additionaly, the expression of Bcl-2 protein increased in E2F1 shRNA1-modified cell line. Conclusions E2F1 is highly expressed in H446 small cell lung cancer cell line. E2F1 promotes apoptosis of H446 through upregulating Bcl-2 expression.

Strict regulation of HIV-1 PR function is critical for efficient production of mature viral particles. During viral protein expression and viral assembly, HIV-1 PR located within Gag-Pol precursor must be inactive to prevent premature cytoplasmic processing of the viral Gag and Gag-Pol precursors. Premature activation of HIV-1 precursors leads to major defects in viral assembly and production of viral particles. A cell-level premature activation of HIV-1 precursors assay using bioluminescence resonance energy transfer (BRET) was established. Three thousand compounds were screened to evaluate this assay. The results showed that the assay is sensitive, specific and stable (Z' factor is 0.905).

The HIV-1 Rev protein facilitates nuclear export of unspliced and singly spliced viral transcripts containing RRE RNA through the CRM1 export pathway. Inhibition of Rev-mediated RNA nuclear export can arrest HIV-1 transcriptional process, which clearly, reveals a target for anti-HIV drug development. In this work, a cell-based assay has been established for screening anti-HIV compounds targeting the Rev-mediated RNA nuclear export. This assay utilized a codon-optimized green fluorescent protein (GFP) as reporter gene, which expression is in a Rev-dependent manner. Any compound that inhibits the Rev-mediated RNA nuclear export is identified by reducing emission of GFP. The Z' score of this model is 0.8220. Three thousands compounds were screened and the positive rate was 9.3% with a cutoff at 50% inhibition. IMB7C7, one of the positive compounds, efficiently inhibits viral production from HIV-1 infected cells.

To establish a cell-based rapid luciferase suppression assay for high-throughput screening (HTS) anti-alphaviruses compounds screening, which could cause viral encephalitis, raise the social issues associated directly with public health and huge economic burden to the society. The Gaussia luciferase assay system was used for HTS model for identifying inhibitors of labeled virus XJ160-GLUC. The decreased 50% GLUC activity inhibition ratio was deemed to be the screening positive index. The reaction system in this model was optimized, and the reliability of the model was evaluated. For HTS model's optimization, cells were infected with XJ160-GLUC at an MOI of 0.025 PFU/cell. The supernatant treated with compounds 48h were collected for GLUC expression detection. In the model, Z' factor was up to 0.71, demonstrating that HTS assay for identifying inhibitors that target all aspects of the viral life cycle of XJ160-GLUC was stable and reliable. After screening 8080 compounds (five-in-one), 341 positive samples were selected, and the positive rate was 4.2% with a cutoff at 50% inhibition. Then 1705 compounds were screened subsequently and the positive rate was 1.1% with obtaining 19 positive compounds. These results will lay the foundation for finding the anti-alphaviruses' drug targets.
Alphavirus ; drug effects ; genetics ; metabolism ; Animals ; Antiviral Agents ; pharmacology ; Drug Evaluation, Preclinical ; methods ; Genes, Reporter ; High-Throughput Screening Assays ; methods ; Luciferases ; genetics ; metabolism

Human immunodeficiency virus (HIV)-1 infection changes transcriptional profiles and regulates. the factors and machinery of the host that facilitate viral replication. Our previous study suggested that the serine/threonine kinase citron kinase (citK) promotes HIV-1 egress. To ascertain if HIV-1 infection affects citK expression in primary cells, peripheral blood mononuclear cells were infected with vesicular stomatitis virus G protein (VSV-G)-pseudotyped HIV-1 vector NL4-3-luc viruses, which resulted in remarkably increased expression of citK. citK overexpression led to a more than two-fold increase in HIV-1 production, whereas a significant decrease was observed when citK was depleted in CD4+ T cells. Infection with HIV-1 pseudoviruses induced increases in the mRNA and protein levels of citK by 2. 5- and 2. 7-fold in HEK293T cells, respectively. By cloning the 5-kb promoter of citK into a luciferase reporter system and transfecting the construct into HEK293T cells, enhanced luciferase activity was observed during HIV-1 infection. Taken together, these data demonstrate that HIV-1 infection upregulates citK expression at the transcriptional level, and thereby renders the host more susceptible to invasion by HIV-1.
CD4-Positive T-Lymphocytes ; virology ; Cloning, Molecular ; Gene Expression Regulation, Enzymologic ; HEK293 Cells ; HIV-1 ; physiology ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Protein-Serine-Threonine Kinases ; genetics ; Up-Regulation ; Virus Replication