1.Function of ubiquitin-specific protease 47 in regulating rabies virus infection
Yannan ZHANG ; Mengyang WU ; Chongyang WANG ; Haomiao DONG ; Xin GUO ; Yidi GUO ; Maolin ZHANG
Chinese Journal of Veterinary Science 2025;45(2):266-273
This study aims to inquire about the fluctuations of ubiquitin-specific protease 47 on neu-roblastoma cells(Neuro-2a,N2a)infected by rabies virus(RABV).USP47 expression levels were detected after RABV infection in N2a cells through RT-qPCR,protein immunoblotting,and virus titer determination.The levels of RABV nucleoprotein and phosphoprotein gene and protein,and RABV titers in supernatants were analyzed during overexpression and knockdown of USP47.The results showed that RABV infection increased USP47 gene level in N2a cells.When overexpression of USP47,the levels of RABV N and P were increased,and the virus titers were also improved.Mo-reover,the level of interleukin-6(IL-6)genes decreased.Knocking down USP47 expression reduced levels of RABV N and P genes and proteins,lowered the virus titer,and elevated the IL-6 gene lev-el.The results suggest that USP47 promotes RABV infection and suppresses IL-6 expression.This finding lays the foundation for further investigation into the molecular mechanisms by which USP47 regulates RABV infection.
2.Function of ubiquitin-specific protease 47 in regulating rabies virus infection
Yannan ZHANG ; Mengyang WU ; Chongyang WANG ; Haomiao DONG ; Xin GUO ; Yidi GUO ; Maolin ZHANG
Chinese Journal of Veterinary Science 2025;45(2):266-273
This study aims to inquire about the fluctuations of ubiquitin-specific protease 47 on neu-roblastoma cells(Neuro-2a,N2a)infected by rabies virus(RABV).USP47 expression levels were detected after RABV infection in N2a cells through RT-qPCR,protein immunoblotting,and virus titer determination.The levels of RABV nucleoprotein and phosphoprotein gene and protein,and RABV titers in supernatants were analyzed during overexpression and knockdown of USP47.The results showed that RABV infection increased USP47 gene level in N2a cells.When overexpression of USP47,the levels of RABV N and P were increased,and the virus titers were also improved.Mo-reover,the level of interleukin-6(IL-6)genes decreased.Knocking down USP47 expression reduced levels of RABV N and P genes and proteins,lowered the virus titer,and elevated the IL-6 gene lev-el.The results suggest that USP47 promotes RABV infection and suppresses IL-6 expression.This finding lays the foundation for further investigation into the molecular mechanisms by which USP47 regulates RABV infection.
3.Phantom study of the influence of iterative algorithm on image quality in ultra-low-dose CT scan of lung
Changjiu HE ; Peng ZHOU ; Shibei HU ; Haomiao QING ; Xiaolei DONG ; Hailie LI ; Deshan LI ; Haipiao LONG
Chinese Journal of Radiological Medicine and Protection 2019;39(2):145-149
Objective To explore the influence of iterative algorithm on image quality in ultralow-dose CT scan of lung.Methods The thoracic model was scanned using different parameter combinations.The tube voltage was chose with 80,100 kV in ultra-low dose group and the mAs was selected by 10,15,20,25 and 30 mAs.The control group selected the parameters of 120 kV,30 mAs.All the images were reconstructed with filtered back projection (FBP group) and iterative algorithm (IR group).The image noise and effective dose (E) were compared.Results When tube current and voltage were constant,the image noise of IR group was lower than that of FBP group,and the difference was statistically significant (t =1.102-8.070,P<0.05).When the tube current was constant,the image noise of the 80 kV with FBP group was higher than that of 100 kV with FBP group,and the image noise of the 80 kV with IR group was lower than the 100 kV with FBP group(t =-8.639-7.841,P<0.05).Compared with the conventional low-dose with FBP group,the image noise of each ultra-low-dose with FBP group was significantly increased,and the image noise of (80 kV,10 mAs),(80 kV,15 mAs),(80 kV,20 mAs) with IR group was significantly increased,and the image noise of (100 kV,15 mAs),(100 kV,20 mAs),(100 kV,25 mAs),(100 kV,30 mAs) with IR group was significantly reduced (t=-8.140-23.028,P<0.05).There was no significant difference in image noise between the groups of (80 kV,25 mAs),(80 kV,30 mAs),(100 kV,10 mAs) with IR and the group of conventional low dose with FBP (P>0.05).Compared with the conventional low dose group,the E of the groups of (80 kV,25 mAs),(80kV,30mAs),(100kV,10 mAs),(100kV,15mAs),(100kV,20mAs),(100kV,25 mAs),(100 kV,30 mAs) was decreased by 75.9%,71.0%,79.8%,70.4%,60.3%,50.2%,40.0%,respectively.Conclusions The image quality of the ultra-low dose protocol (100 kV,10mAs) with iterative algorithm is similar to that of the conventional low dose with FBP group,and the radiation dose could be significantly reduced.
4.Cloning, expression and characterization of beta-glucosidase from Aspergillus fumigatus.
Yi XIE ; Haomiao OUYANG ; Ribo HUANG ; Dong CHEN ; Cheng JIN
Chinese Journal of Biotechnology 2013;29(9):1245-1253
Exploring new beta-glucosidase genes is of great importance to industrialize beta-glucosidase. The genomes of Aspergillus fumigatus contain a bgl gene, which encodes a 65 kDa putative beta-glucosidase. The bgl gene was cloned into an expression plasmid and transformed to Escherichia coli BL21 (DE3). The bgl was expressed upon induction of Isopropyl beta-D-1-thiogalactopyranoside (IPTG). The recombinant protein was purified by GST-tag affinity chromatography. The purified recombinant Bgl was characterized using Esculin as substrate. The optimum temperature and pH were 45 degrees C and 5.0-6.0, respectively. The K(m) for Esculin was 17.7 mmol/L. The enzyme was stable in the range of pH 4-7. After incubation at 70 degrees C for 2 h, the recombinant Bgl remained 60% of its activity. Metal ions and chemical reagents had different influences on the activity of beta-glucosidase. Ca2+ (1 mmol/L) could increase enzyme activity slightly. On the contrary, the enzyme activity was greatly inhibited by 5 mmol/L Sodium dodecyl sulfate (SDS). Based on our results, the A. fumigatus Bgl was thermostable beta-glucosidase.
Aspergillus fumigatus
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enzymology
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Cloning, Molecular
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Enzyme Stability
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Escherichia coli
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genetics
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metabolism
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Recombinant Proteins
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biosynthesis
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genetics
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metabolism
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beta-Glucosidase
;
biosynthesis
;
genetics
;
metabolism

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