Research progress of natural products against hepatitis C virus / 药学实践杂志

Hepatitis C virus (HCV) is an important factor leading to chronic hepatitis, cirrhosis and hepatocellular carcinoma. Currently, there is no vaccine available to prevent HCV infection. This article reviews the natural products with anti-HCV activity discovered from plants in recent years, and classifies and summarizes them according to their mechanism of action, in order to provide a basis for discovering anti-HCV drugs from natural products.

Defeating enemy outside the border-development of viral entry inhibitors / 复旦学报（医学版）

About 25 years ago,we had pioneered the research fields of developing human immunodeficiency virus (HIV) fusion/entry inhibitors and anti-HIV peptide drugs.Over the past six years,we have gained some promising results in research and development of the HIV,Middle East respiratory syndrome coronavirus (MERS-COV),the Ebola virus (EBOV),and the Zika virus (ZIKV) entry inhibitors.This article provides an overview of the research progress of viral entry inhibitors against the related highly pathogenic viruses.

Amelioration of experimental autoimmune myocarditis by HVEM-overexpressing dendritic cells through induction of IL-10-producing cells / 中华微生物学和免疫学杂志

ObjectiveTo assess the efficacy of herpes virus entry mediator (HVEM) gene modifled dendritic cells (DCs) in protecting against myosin induced myocarditis,and to investigate the involving mechanism.MethodsWe treated experimental autoimmune myocarditis (EAM) mice with myosin-pulsed DCs which were transfected with HVEM-expressing adenovirus (Ad-HVEM) or control vectors,then evaluated myocarditis,plasm cTn [ and autoantibody by histopathology,fluoroimmunoassay,and ELISA,respectively.ResultsWe found that DCs transfected with Ad-HVEM (DC-Ad-HVEM) could protect against EAM.Further study showed DC-Ad-HVEM could produce regulatory cytokine IL-10,and IL-10 promoted the production of a key regulatory T cell subset which is important in peripheral tolerance.The T cells mediated protection against EAM.ConclusionThis study suggest that myosin-DC-Ad-HVEM cell gene therapy is a safe and effective way for inhibiting the development of EAM,and the signal net mediated by HVEM plays different roles in different cells.

Effect of aldosterone on the amplification of oncolytic vaccinia virus in human cancer lines / 대한간학회지

BACKGROUND/AIMS: JX-594 is an oncolytic virus derived from the Wyeth vaccinia strain that causes replication-dependent cytolysis and antitumor immunity. Starting with a cross-examination of clinical-trial samples from advanced hepatocellular carcinoma patients having high levels of aldosterone and virus amplification in JX-594 treatment, we investigated the association between virus amplification and aldosterone in human cancer cell lines. METHODS: Cell proliferation was determined by a cell-counting-kit-based colorimetric assay, and vaccinia virus quantitation was performed by quantitative polymerase chain reaction (qPCR) and a viral plaque assay. Also, the intracellular pH was measured using a pH-sensitive dye. RESULTS: Simultaneous treatment with JX-594 and aldosterone significantly increased viral replication in A2780, PC-3, and HepG2 cell lines, but not in U2OS cell lines. Furthermore, the aldosterone treatment time altered the JX-594 replication according to the cell line. The JX-594 replication peaked after 48 and 24 hours of treatment in PC-3 and HepG2 cells, respectively. qPCR showed that JX-594 entry across the plasma membrane was increased, however, the changes are not significant by the treatment. This was inhibited by treatment with spironolactone (an aldosterone-receptor inhibitor). JX-594 entry was significantly decreased by treatment with EIPA [5-(N-ethyl-N-isopropyl)amiloride; a Na+/H+-exchange inhibitor], but aldosterone significantly restored JX-594 entry even in the presence of EIPA. Intracellular alkalization was observed after aldosterone treatment but was acidified by EIPA treatment. CONCLUSIONS: Aldosterone stimulates JX-594 amplification via increased virus entry by affecting the H+ gradient.

Clone and expression of human cytomegalovirus UL144 gene and its effects on dendritic cells / 第二军医大学学报

Objective: To construct a recombinant adenovirus vector carrying human cytomegalovirus (hCMV) UL144 gene and to explore the biological characteristics of UL144 gene-modified DCs. Methods: The UL144 gene was amplified from hCMV DNA, which was extracted from hCMV-DNA positive serum. The recombinant adenovirus vector carrying hCMV UL144 gene was constructed with AdEasy system and then transfected into HEK293 cells to create recombinant adenovirus Ad-UL144. The expression of inserted gene was identified by RT-PCR. The recombinant adenovirus was then transfected into mice myeloid dendritic cells. The surface proteins of dendritic cells were analyzed by FACS, and cytokines in supernatant were detected by ELISA. T cell proliferation stimulated by gene-modified DC was examined by 3H-TdR uptake assay. Results: The UL144 gene was successfully cloned into the pAdEasy-1 plasmid. The recombinant adenovirus Ad-UL144 was packed in HEK293 cells, with a viral titer of 3×10 10 pfu/ml. DCs infected with AdCMV-UL144 had markedly decreased surface expression of CD80, CD86 and I-Ad (P < 0.01). The contents of TNF-α, IL-6 and IL-1β were significantly decreased in the supernatant of AdCMV-UL144 modified DCs (P < 0.05). T cell proliferation ability induced by gene-modified DC was obviously lower than in the DC control group (P < 0.01). Conclusion: UL144-modified DCs can maintain a relative immature status, and have reduced stimulating activity upon the proliferation and activation of T cells in vitro.

Recent Advances in Hepatitis B Virus Entry Research / 生物化学与生物物理进展

The mechanism of hepatitis B virus entry is an interesting area in HBV research but still enigmatic.The difficulties in HBV entry research were primarily caused by the lack of easily accessible in vitro infection models.Recent years,primary hepatocytes from Tupaia belangeri has been substituted for primary human hepatocytes and upon induction of differentiation in vitro.A human hepatoma cell line named HepaRG has been found to be susceptible for HBV infection too.The two cell models enabled researchers to obtain a number of important discoveries for HBV entry.This article are focusing on these discoveries,including the domains of HBV surface proteins involved in HBV entry,potential HBV receptor candidates and the questions to be resolved in future years.

Clone and expression of human cytomegalovirus UL144 gene and its effects on dendritic cells / 第二军医大学学报

Objective:To construct a recombinant adenovirus vector carrying human cytomegalovirus(hCMV) UL144 gene and to explore the biological characteristics of UL144 gene-modified DCs.Methods:The UL144 gene was amplified from hCMV DNA,which was extracted from hCMV-DNA positive serum.The recombinant adenovirus vector carrying hCMV UL144 gene was constructed with AdEasy system and then transfected into HEK293 cells to create recombinant adenovirus Ad-UL144.The expression of inserted gene was identified by RT-PCR.The recombinant adenovirus was then transfected into mice myeloid dendritic cells.The surface proteins of dendritic cells were analyzed by FACS,and cytokines in supernatant were detected by ELISA.T cell proliferation stimulated by gene-modified DC was examined by 3H-TdR uptake assay.Results:The UL144 gene was successfully cloned into the pAdEasy-1 plasmid.The recombinant adenovirus Ad-UL144 was packed in HEK293 cells,with a viral titer of 3?1010 pfu/ml.DCs infected with AdCMV-UL144 had markedly decreased surface expression of CD80,CD86 and I-Ad(P