1.Silencing of UBP43 by shRNA Enhances the Antiviral Activity of Interferon against Hepatitis B Virus
Hebin FAN ; Baoju WANG ; Yinping LU ; Youhua HAO ; Xinxing YANG ; Mengji LU ; Dongliang YANG
Virologica Sinica 2008;23(5):339-344
Previous studies have shown that expression of the interferon-sensitive gene (ISG)I5 protease UBP43 is increased in the liver biopsy specimens of patients who do not respond to interferon (IFN)-α therapy. We hypothesized that UBP43 might hinder the ability of IFN to inhibit HBV replication. In this study, we investigated whether vector-based siRNA promoted by Hi (psiUBP43) could enhance IFN inhibiting HBV replication in cell culture. UBP43 was specifically silenced using shRNA. In HepG2.2.15 cells, the HBeAg and HBV DNA levels were significantly reduced by IFN after transfection of shRNA, imphicated that vector-based siRNA promoted by HI (psiUBP43) could enhance IFN inhibiting HBV replication in cell culture. These data suggest that UBP43 modulates the anti-HBV type I IFN response, and is a possible therapeutic target for the treatment of HBV infection.
2.Risk factors for early miscarriage among intrauterine singleton pregnancies after treatment with in vitro fertilization/intracytoplasmic sperm injection.
Hong ZENG ; Nenghui LIU ; Xiuli FAN ; Muzhu CAI ; Hebin XIE
Journal of Central South University(Medical Sciences) 2016;41(8):815-820
OBJECTIVE:
To investigate the risk factors associated with early miscarriage among intrauterine singleton pregnancies after treatment with in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI).
METHODS:
A retrospective case-control study was performed on all singleton pregnancies underwent IVF/ICSI from January, 2013 to May, 2014, in Xiangya Hospital. Ninety-six early miscarriage patients served as a case group and 593 pregnancies with live birth served as a control group. We analyzed factors for early miscarriage after IVF/ICSI in two groups.
RESULTS:
Multivariate logistic regression analysis demonstrated that the women age, miscarriage history, and sperm DAN fragmentation index (DFI) were the risk factors for early miscarriage (P<0.05).
CONCLUSION
Miscarriage after treatment with IVF/ICSI is affected by multiple factors. Women at elder age (>30 years old), women with a history of miscarriage or men with higher sperm DFI (≥15%) are the risk.
Abortion, Spontaneous
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Case-Control Studies
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Female
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Fertilization in Vitro
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Humans
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Male
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Pregnancy
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Retrospective Studies
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Risk Factors
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Sperm Injections, Intracytoplasmic
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Spermatozoa
3.Preliminary identification of the cloning, expression, and function of Marmota himalayana type I interferon receptor β subunit
Ying TAO ; Dongliang YANG ; Baoju WANG ; Yi LIU ; Wenjia GUI ; Zhi LI ; Hebin FAN
Journal of Clinical Hepatology 2024;40(2):278-283
ObjectiveTo clone the gene of Marmota himalayana type Ⅰ interferon receptor β subunit (mhIFNAR2), and to perform antibody preparation and functional identification. MethodsRT-PCR was used for amplification in the spleen tissue of Marmota himalayana to obtain the sequence, which was cloned to the prokaryotic expression vector pRSET-B to express the recombinant protein. Electrophoresis and Western blot were used for identification. BALB/c mice were immunized with the recombinant protein to prepare the polyclonal antibody of its extracellular domain; immunohistochemistry, immunofluorescence assay, and Western Blot were used for identification, and the method of siRNA blockade was used to investigate its function. An analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for comparison between two groups. ResultsA fragment of mhIFNAR2 (149 — 1 300 bp) was obtained from spleen tissue, which showed the highest homology of 98.05% in marmot. A prokaryotic expression plasmid was successfully constructed for expression of the extracellular domain of the mhIFNAR2(50-181aa) and was named pRSET-B.mhIFNAR2, and the recombinant protein expressed by this plasmid had a molecular weight of 27 kD, a purity of about 95% after purification, and a concentration of 160 μg/mL. After BALB/c mice were immunized with the purified recombinant protein, 1∶1 000 specific polyclonal antibodies were obtained, and immunohistochemistry and immunofluorescence assay showed the expression in cell membrane and cytoplasm. Among the three siRNAs synthesized, the siRNA starting from the 277 locus (siRNA277) could silence the expression of target genes and weaken the interferon signaling pathway compared with the blank control group and the negative control group (both P<0.05). ConclusionThe fragment of mhIFNAR2 is obtained, and the polyclonal antibody for the extracellular domain of mhIFNAR2 is successfully prepared, with relatively high titer and specificity, and can be used for immunohistochemistry, immunofluorescence assay, and Western blot.