1.Construction and immune efficacy evaluation of BNeV VLPs based on VP1 protein in mice
Lu DING ; Xiangyue HUANG ; Jinbo WU ; Chaohui ZHANG ; Qing ZHU ; Chenxi ZHU ; Gu-nan DENG ; Ajia AKE ; Chunsai HE ; Yuanzhen MA ; Bin ZHANG
Chinese Journal of Veterinary Science 2025;45(3):412-419
The codon was optimized for the bovine nebovirus(BNeV)VP1 gene and the recombi-nant plasmid pFastBac-Dual-VP1 was constructed,and BNeV-VP1 virus-like particles(VLPs)were prepared using a baculovirus expression system,and identified by Western blot,indirect im-munofluorescence and electron microscopy.Successfully validated VLPs were mixed with MF59 adjuvant and CpG-ODG,and mice were immunised by intramuscular injection and evaluated for immunity effects.The results showed that the optimized CAI(codon adaptation index)of VP1 gene was 0.93 and the GC content was 60.4%.The constructed recombinant plasmid was trans-formed into DH10Bac for blue-white spot screening,and after successful verification,it was trans-fected into SF9 cells to obtain recombinant baculovirus Baculo-BNeV-VP1.BNeV virus-like parti-cles with diameters ranging from 35 to 40 nm were observed under the electron microscope,and both IFA and Western blot experiments proved that the target proteins were successfully ex-pressed and biologically active,and protein optimisation revealed that the highest protein expres-sion was found at the infectious dose MOI=0.5.Mice were immunized by intramuscular injection after 50 μg of VLPs were mixed with MF59 adjuvant and CpG-ODN.The results showed that the VLPs immunization group produced IgG antibodies 7 days after the first dose,and the antibody ti-ter increased gradually,reaching a maximum of 1∶102 400,and declined at 35 d,but still main-tained a high level;The blocking titer BT50 is up to 640,which can induce the production of BNeV VP1-specific blocking antibody in mice.In this study,the baculovirus expression system was used to express the VP1 protein of BNeV,and BNeV VLPs were successfully constructed,which could induce humoral immune response in mice,which provided a reference for the follow-up research of BNeV vaccine.
2.Preparation and immune efficacy evaluation of bovine parainfluenza type 3 virus like particles
Chenxi ZHU ; Xiangyue HUANG ; Qing ZHU ; Lu DING ; Gunan DENG ; Ajia AKE ; Chunsai HE ; Yuanzhen MA ; Jinbo WU ; Chaohui ZHANG ; Bin ZHANG
Chinese Journal of Veterinary Science 2025;45(3):404-411,442
Codon optimization was performed for the M and HN genes of bovine parainfluenza virus type 3(BPIV3),and the recombinant shuttle plasmid Dual-M+HN was constructed.BPIV3 VLPs was prepared using the baculovirus expression system,and verified by Western blot,IFA and elec-tron microscopy.The successfully verified virus-like particle(VLPs)were mixed with MF59 adjuvant and CpG-ODN immunoenhancer to immunize mice by intramuscular-injection,and BPIV3 inactivated vaccine group and adjuvant control group were set up.The immune effect of BPIV3 VLPs was evaluated by monitoring mouse serum specific antibodies,neutralizing antibodies and hemagglutination inhibition antibodies.The results showed that the optimized codon adaptation in-dex(CAI)of the M and HN protein genes were 0.96 and 0.95,respectively,and the CG content reached 54.1%and 53.1%,respectively.The constructed recombinant plasmid was transformed in-to DHI0Bac for blue and white spot screening.The validated recombinant rod particles were trans-fected into Sf9 cells to obtain the rod-shaped virus pFastBac-M+HN.Under electron microscopy,BPIV3 VLPs with a diameter of approximately 180 nm were observed.IFA and Western blot ex-periments confirmed the successful expression and biological activity of the target protein.Through protein optimization,it was found that the protein expression was highest at an infection dose of MOI=5.After mixing 50 μg VLPs with MF59 adjuvant and CpG-ODN,mice were immunized by intramuscular injection.The results showed that the antibodies in the VLPs immunized group be-gan to rise at 2 weeks of the first immunization and reached their peak at 21 days of the second im-munization,with an average IgG antibody titer of 1∶40 228;The average titer of neutralizing anti-bodies is 1∶298;The titer of hemagglutination inhibition antibody is 1∶549,reaching the level of inactivated vaccine(P≥0.05),indicating that the VLPs prepared in this experiment can induce hu-moral immune response in the body.In summary,this study successfully prepared VLPs capable of self-assembly expression of BPIV3 HN and M proteins,and induced humoral immune response in mice,providing research basis for subsequent BPIV3 VLPs vaccine research.
3.Construction and immune efficacy evaluation of BNeV VLPs based on VP1 protein in mice
Lu DING ; Xiangyue HUANG ; Jinbo WU ; Chaohui ZHANG ; Qing ZHU ; Chenxi ZHU ; Gu-nan DENG ; Ajia AKE ; Chunsai HE ; Yuanzhen MA ; Bin ZHANG
Chinese Journal of Veterinary Science 2025;45(3):412-419
The codon was optimized for the bovine nebovirus(BNeV)VP1 gene and the recombi-nant plasmid pFastBac-Dual-VP1 was constructed,and BNeV-VP1 virus-like particles(VLPs)were prepared using a baculovirus expression system,and identified by Western blot,indirect im-munofluorescence and electron microscopy.Successfully validated VLPs were mixed with MF59 adjuvant and CpG-ODG,and mice were immunised by intramuscular injection and evaluated for immunity effects.The results showed that the optimized CAI(codon adaptation index)of VP1 gene was 0.93 and the GC content was 60.4%.The constructed recombinant plasmid was trans-formed into DH10Bac for blue-white spot screening,and after successful verification,it was trans-fected into SF9 cells to obtain recombinant baculovirus Baculo-BNeV-VP1.BNeV virus-like parti-cles with diameters ranging from 35 to 40 nm were observed under the electron microscope,and both IFA and Western blot experiments proved that the target proteins were successfully ex-pressed and biologically active,and protein optimisation revealed that the highest protein expres-sion was found at the infectious dose MOI=0.5.Mice were immunized by intramuscular injection after 50 μg of VLPs were mixed with MF59 adjuvant and CpG-ODN.The results showed that the VLPs immunization group produced IgG antibodies 7 days after the first dose,and the antibody ti-ter increased gradually,reaching a maximum of 1∶102 400,and declined at 35 d,but still main-tained a high level;The blocking titer BT50 is up to 640,which can induce the production of BNeV VP1-specific blocking antibody in mice.In this study,the baculovirus expression system was used to express the VP1 protein of BNeV,and BNeV VLPs were successfully constructed,which could induce humoral immune response in mice,which provided a reference for the follow-up research of BNeV vaccine.
4.Preparation and immune efficacy evaluation of bovine parainfluenza type 3 virus like particles
Chenxi ZHU ; Xiangyue HUANG ; Qing ZHU ; Lu DING ; Gunan DENG ; Ajia AKE ; Chunsai HE ; Yuanzhen MA ; Jinbo WU ; Chaohui ZHANG ; Bin ZHANG
Chinese Journal of Veterinary Science 2025;45(3):404-411,442
Codon optimization was performed for the M and HN genes of bovine parainfluenza virus type 3(BPIV3),and the recombinant shuttle plasmid Dual-M+HN was constructed.BPIV3 VLPs was prepared using the baculovirus expression system,and verified by Western blot,IFA and elec-tron microscopy.The successfully verified virus-like particle(VLPs)were mixed with MF59 adjuvant and CpG-ODN immunoenhancer to immunize mice by intramuscular-injection,and BPIV3 inactivated vaccine group and adjuvant control group were set up.The immune effect of BPIV3 VLPs was evaluated by monitoring mouse serum specific antibodies,neutralizing antibodies and hemagglutination inhibition antibodies.The results showed that the optimized codon adaptation in-dex(CAI)of the M and HN protein genes were 0.96 and 0.95,respectively,and the CG content reached 54.1%and 53.1%,respectively.The constructed recombinant plasmid was transformed in-to DHI0Bac for blue and white spot screening.The validated recombinant rod particles were trans-fected into Sf9 cells to obtain the rod-shaped virus pFastBac-M+HN.Under electron microscopy,BPIV3 VLPs with a diameter of approximately 180 nm were observed.IFA and Western blot ex-periments confirmed the successful expression and biological activity of the target protein.Through protein optimization,it was found that the protein expression was highest at an infection dose of MOI=5.After mixing 50 μg VLPs with MF59 adjuvant and CpG-ODN,mice were immunized by intramuscular injection.The results showed that the antibodies in the VLPs immunized group be-gan to rise at 2 weeks of the first immunization and reached their peak at 21 days of the second im-munization,with an average IgG antibody titer of 1∶40 228;The average titer of neutralizing anti-bodies is 1∶298;The titer of hemagglutination inhibition antibody is 1∶549,reaching the level of inactivated vaccine(P≥0.05),indicating that the VLPs prepared in this experiment can induce hu-moral immune response in the body.In summary,this study successfully prepared VLPs capable of self-assembly expression of BPIV3 HN and M proteins,and induced humoral immune response in mice,providing research basis for subsequent BPIV3 VLPs vaccine research.
5.Clinical Significance and Method of Measuring Lung Fluid Viscosity and Surface Tension
Yuanyang LIU ; Zhenglong CHEN ; Yuanlin SONG ; Xiangyue HUANG ; Chunyuan ZHANG ; Zhaoyan HU
Journal of Medical Biomechanics 2020;35(3):E378-E384
Pulmonary fluid refers to a thin and continuous liquid layer mainly consisting of airway mucus and lining liquid on the inner surface of alveoli, which is essential for maintaining a healthy lung. The clinical significance of rheological properties of pulmonary fluid in airway stability, pulmonary barrier and clearance functions, ventilator induced lung injury and surfactant replacement therapy in infant respiratory distress syndrome was introduced in this paper. Furthermore, a variety of classical methods for measuring surface tension and liquid viscosity, such as Langmuir-Wilhelmy balance, captive bubble method, glass capillary viscometer and rotational viscometer, and emerging techniques (e.g. particle tracking microrheometer and axisymmetric drop shape analysis) were reviewed, and their advantages and drawbacks were also compared, in order to provide an important reference for the assisted diagnosis and treatment of lung diseases in clinical practice.
6.A single person operation procedure and experience of tail vein injection in conscious tree shrews
Yongwang HE ; Juan LI ; Jian HUANG ; Yiqing ZHOU ; Xiangyue ZHAO ; Genmeng YANG ; Zhen LI ; Na LI ; Xiaofeng ZENG
Chinese Journal of Comparative Medicine 2018;28(3):91-94
Objective To introduce a single person operation procedure for tail vein injection in conscious tree shrews,and to improve the success rate of injection. Methods The tree shrew was fixed by a canvas glove and a clamp. The tail of the tree shrew was fixed by the operator's left hand, and the drug was injected by the right hand with a 1 mL disposable syringe. Results This method had the advantages of simple operation,economy and practicality,good fixation effect,high matching degree of animals and high success rate of drug administration. Conclusions Compared with other methods,this method has obvious advantages such as single person operation, convenient, time-saving and labor-saving. The cost is low and the effect is good,thus significantly improving the success rate of injection.
7.Behavior research of allergic rhinitis with adenoid hypertrophy in children.
Yunlong JING ; Binya HU ; Jing HUANG ; Sijun ZHAO ; Lihua TAO ; Xiangyue PENG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2012;26(12):547-549
OBJECTIVE:
To investigate the behavior difference of allergic rhinitis with adenoid hypertrophy between study group and control group.
METHOD:
One hundred and seventeen children diagnosed as allergic rhinitis with adenoid hypertrophy were enrolled in our study were divided into study group and control group. Forty-two children treated with local steroid nasal spray for two to three months and antihistamine were control group. Seventy-five children treated with endoscopic adenoidectomy and drug treatment were study group; All children' parents were inquired for their clinical presentation.
RESULT:
No distinctive differences were found between the two groups (P > 0.05) in adenoid hypertrophy, accompanying nasal problems and clinical questionnaire scoring. Significant statistical distinction were found (P < 0.05) in snoring, sleep disturbance and frequent arousal, nasal obstructive moth-breathing, and recurrent respiratory tract infection between the two groups after three-month follow up.
CONCLUSION
Endoscopic adenoidectomy should be taken into account for allergic rhinitis with adenoid hypertrophy in children. Adenoidectomy would be useful for the improvement of behavior symptoms.
Adenoidectomy
;
methods
;
Adenoids
;
pathology
;
Anti-Allergic Agents
;
administration & dosage
;
Case-Control Studies
;
Child
;
Combined Modality Therapy
;
Endoscopy
;
Female
;
Histamine H1 Antagonists
;
administration & dosage
;
Humans
;
Hypertrophy
;
complications
;
therapy
;
Male
;
Rhinitis, Allergic
;
complications
;
therapy
;
Steroids
;
administration & dosage
8.The effect of tympanostomy tube surgery in cleft palate children with secretory otitis media.
Min HUANG ; Sijun ZHAO ; Yun LI ; Xiangyue PENG ; Yuting KUANG ; Songliang LONG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2012;26(22):1017-1019
OBJECTIVE:
To study the effect of tympanostomy tube surgery in cleft palate children with secretory otitis media (SOM).
METHOD:
Cleft palate children complicated by SOM accepted palatoplasty combined tympanostomy tube surgery, and evaluated by tympanum examination, acoustic immittance measurement in the post-operation. The others with suited age constitute, who accepted palatoplasty only,received the same testing too.
RESULT:
Compared with children who accepted palatoplasty only,the correlated index of SOM in post-operation of children who accepted palatoplasty combined with tympanostomy tube surgery had more improvements,there was a statistically significant difference between them. The recovery degree of middle ear function in younger group was better than elder group,there was a statistically significant difference between them.
CONCLUSION
Tympanostomy tube surgery is helpful to increase cure rate of 0ME in children with cleft palate. The younger the patient is,the better the recovery degree of middle ear function will be. Palatoplasty combined with tympanostomy tube surgery should be an effective way in the management of cleft palate complicated by SOM, and good effect could be obtained when children accepted operation in early stage.
Age Factors
;
Child
;
Child, Preschool
;
Cleft Palate
;
surgery
;
Humans
;
Infant
;
Middle Ear Ventilation
;
Otitis Media with Effusion
;
surgery

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