1.Development and evaluation of immunoprotective efficacy of a virus-like particle vaccine against encephalomyocarditis virus
Yanfang ZHANG ; Qiong ZHU ; Jie FU ; Yaohui FANG ; Jiayin JIN ; Danna ZHANG ; Fei DENG ; Shengbo CAO
Chinese Journal of Veterinary Science 2025;45(5):994-1001
Encephalomyocarditis virus(EMCV)is a zoonotic pathogen that causes encephalitis and myocarditis as its primary clinical manifestations.To explore effective preventive measures,this study utilized a Bac-to-Bac expression system to insert the EMCV P12A and 3C genes into the pFastBacDual shuttle vector,resulting in the generation of the recombinant baculovirus Ac-P12A-3C.This facilitated the large-scale expression and purification of EMCV virus-like particles(VLPs),which were correctly assembled into particles of approximately 30 nm in diameter,as ob-served by electron microscopy.Immunization and challenge experiments in mice demonstrated that these VLPs could effectively protect against EMCV infection,achieving a protection rate of 100%.Histopathological sections indicated that,compared to the PBS control group,the VLP immuniza-tion group exhibited significantly reduced tissue damage,along with a marked decrease in viral load within the tissues.In piglets,immunization with the VLPs elicited a robust humoral response,with neutralizing antibody titers reaching 1∶320 to 1∶640 after a second immunization,and no signifi-cant adverse reactions were observed throughout the immunization process.This study preliminarily explores the immunogenicity and safety of the VLP vaccine,laying the foundation for the development of a subunit vaccine based on EMCV VLPs and offering a new strategy for the prevention and control of encephalomyocarditis.
2.Development and evaluation of immunoprotective efficacy of a virus-like particle vaccine against encephalomyocarditis virus
Yanfang ZHANG ; Qiong ZHU ; Jie FU ; Yaohui FANG ; Jiayin JIN ; Danna ZHANG ; Fei DENG ; Shengbo CAO
Chinese Journal of Veterinary Science 2025;45(5):994-1001
Encephalomyocarditis virus(EMCV)is a zoonotic pathogen that causes encephalitis and myocarditis as its primary clinical manifestations.To explore effective preventive measures,this study utilized a Bac-to-Bac expression system to insert the EMCV P12A and 3C genes into the pFastBacDual shuttle vector,resulting in the generation of the recombinant baculovirus Ac-P12A-3C.This facilitated the large-scale expression and purification of EMCV virus-like particles(VLPs),which were correctly assembled into particles of approximately 30 nm in diameter,as ob-served by electron microscopy.Immunization and challenge experiments in mice demonstrated that these VLPs could effectively protect against EMCV infection,achieving a protection rate of 100%.Histopathological sections indicated that,compared to the PBS control group,the VLP immuniza-tion group exhibited significantly reduced tissue damage,along with a marked decrease in viral load within the tissues.In piglets,immunization with the VLPs elicited a robust humoral response,with neutralizing antibody titers reaching 1∶320 to 1∶640 after a second immunization,and no signifi-cant adverse reactions were observed throughout the immunization process.This study preliminarily explores the immunogenicity and safety of the VLP vaccine,laying the foundation for the development of a subunit vaccine based on EMCV VLPs and offering a new strategy for the prevention and control of encephalomyocarditis.
3.Effect of Pinggan Qianyang recipe on the expression of Tpx II HSP27 and ANXA1 in the hypothalamus of spontaneously hypertensive rats with hyperactivity of liver-YANG syndrome.
Guangwei ZHONG ; Lingli XIANG ; Jianjun HU ; Yaohui YIN ; Qiong CHEN ; Xia FANG
Journal of Central South University(Medical Sciences) 2015;40(2):136-143
OBJECTIVE:
To investigate the effect Pinggan Qianyang recipe on expression of Tpx, HSP27 and ANXA1 in the hypothalamus of spontaneously hypertensive rats (SHRs) with the hyperactivity of liver-YANG syndrome.
METHODS:
A total of 30 SHRs were subjected to administration of Aconiti Praeparatae Decoction to establish the model of SHR with liver-YANG hyperactivity first, then they were randomly divided into three groups: the control group, the model group and the treatment group (n=10 per group). A total of 10 SD rats were served as the normal group. The rats in control group and treatment group were given Enalapril plus Pinggan Qianyang recipe for four weeks. The change of behavior and blood pressure of rats were monitored. RT-PCR and Western-blot were performed to detect the expression of Tpx II, HSP27 and ANXA1 mRNA and protein in the hypothalamus, respectively.
RESULTS:
Compared with the normal SD rats, the heart rate, blood pressure and grade of irritability were significantly increased while rotation endurance time was dramatically reduced in the SHR model with liver-YANG hyperactivity (P<0.01), these changes were reversed by the application of Enalapril plus Pinggan Qianyang recipe. Compared with the normal SD rats, the protein and mRNA expression of Tpx II and ANXA1 in the model group were significantly upregulated (P<0.01) while the HSP27 was significantly downregulated (P<0.01). Compared with the model group, the protein and mRNA expression of Tpx II and ANXA1 in the control group or treatment group were significantly decreased (P<0.05 or P<0.01) while HSP27 was significantly increased (P<0.05 or P<0.01). Compared with the control group, the expression of Tpx II and ANXA1 protein in treatment group were significantly reduced (P<0.05 or P<0.01).
CONCLUSION
Pinggan Qianyang recipe can improve the blood pressure and behavior in SHRs with hyperactivity of Liver-YANG syndrome, which might be related to the regulation of Tpx, HSP27 and ANXA1 expression in hypothalamuses.
Animals
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Annexin A1
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metabolism
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Blood Pressure
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drug effects
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Drugs, Chinese Herbal
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pharmacology
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Enalapril
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pharmacology
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HSP27 Heat-Shock Proteins
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metabolism
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Hypothalamus
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drug effects
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metabolism
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Liver
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physiopathology
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Rats
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Rats, Inbred SHR
4.Magnetic resonance imaging findings of traumatic temporomandibular joint injury induced by type Ⅵ condylar fracture
Yaohui YU ; Meihao WANG ; Dengfeng LIU ; Yiming FANG ; Xinghao ZHU ; Qiping REN ; Lulu PAN
Chinese Journal of Trauma 2012;28(1):46-48
Objective To investigate the application of MRI in evaluation of the traumatic temporomandibular joint (TMJ) injury induced by type Ⅵ condylar fracture. MethodsMRI was performed in TMJs in 18 patients with type Ⅵ condylar fractures at days 3-14 post-injury and the MRI findings were analyzed. ResultsMRI findings of 18 patients with traumatic TMJ injury with 19 sides of type Ⅵ condylar fractures showed 15 sides of TMJ disk displacement,nine sides of capsule tear,16 sides of retrodiscal tissue tear (double-plate area) and 19 sides of joint effusion change. Conclusions MRI is very important in the diagnosis and evaluation of traumatic TMJ injury,since it can clearly display the TMJ injuries in type Ⅵ condylar fractures.Therefore,the clinical application of MRI is beneficial for selection of the therapeutic schedules.
5.Cleavage activity of anti-transforming growth factor ?_1 RNA by U1 snRNA chimeric ribozyme in vitro
Fang LIU ; Ping ZOU ; Yaohui WU ; Min ZHANG
Chinese Journal of Pathophysiology 2000;0(08):-
AIM: To determine the cleavage activity of anti-transforming growth factor ?1 hammerhead ribozymes which was inserted into U1 small nuclear RNA in cell-free system.METHODS: The hammerhead ribozyme targeting against transforming growth factor ?1 was designed through the analysis of computer software.The ribozyme fragments were synthesized and cloned into the U1 snRNA ribozyme vector pZeoU1EcoSpe,which contained U1 snRNA promoter/enhancer and terminator.TGF ?1 cDNA partial fragment was generated by RT-PCR,and then cloned into the T-vector at the downstream of T7 promoter.The transcripts of ribozyme and target RNA incorporated into isotope were transcribed in vitro and purified by denaturing polyacrylamide gel electrophoresis.-labeled U1 snRNA chimeric ribozyme transcripts were incubated with target-RNAs at different conditions and autoradiographed after running denaturing PAGE.RESULTS: U1snRNA chimeric ribozyme(U1Rz803) cleaved TGF?1 mRNA efficiently and specifically at 37 ℃,while the disable ribozyme(U1Rz803m) showed no cleavage activity,so these indicated the design of U1Rz803 was correct.CONCLUSION: U1Rz803 prepared in this study possesses the perfect specific catalytic cleavage activity in cell-free system.These results indicate that U1 snRNA chimeric ribozyme U1Rz803 may suppress the expression of TGF?1 in vivo,therefore it may provide a new means for exploring the role of TGF?1 in hematopoietic regulation in the future.

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