1.Three-dimensional displacement and risk factors of midshaft clavicle fractures treated with titanium elastic intramedullary nailing
Junwei ZHANG ; Lingling CHEN ; Zhenyuan MA ; Weizhi NIE ; Chaohui LI ; Haitao WANG ; Laibao DUAN ; Jinyong HOU ; Hongzheng BI
Chinese Journal of Tissue Engineering Research 2026;30(2):269-277
BACKGROUND:Titanium elastic intramedullary nailing for the treatment of significantly displaced midshaft clavicle fractures has the characteristics of minimally invasive and elastic fixation.The displacement of the fracture is closely related to the later function.However,there are few studies on the three-dimensional displacement analysis of the fracture ends before surgery and after intramedullary fixation such as titanium elastic intramedullary nailing.OBJECTIVE:To explore the three-dimensional displacement of fracture ends after midshaft clavicle fracture and fixation with titanium elastic intramedullary nails,and to analyze the risk factors.METHODS:A total of 91 patients with midshaft clavicle fracture(fracture end shortening ≥15 mm)admitted to Wendeng Orthopedic Hospital of Shandong Province from April 2019 to April 2024 were selected,including 57 males and 34 females,aged(51.73±10.21)years old.All patients received closed reduction and internal fixation with titanium elastic intramedullary nail.CT scans of the affected clavicle were performed before and on the first day after surgery.The CT data were imported into Mimics software for modeling.The length of the clavicle,lateral displacement of the fracture end,and rotation of the distal end of the fracture along the X,Y,and Z axes were measured and recorded before and after surgery.Pearson correlation coefficient was used for correlation analysis of various parameters,and generalized linear regression was used to evaluate risk factors.RESULTS AND CONCLUSION:(1)Preoperatively,the variable that increased the risk of lateral displacement was the number of comminuted bone fragments,the variable that increased the risk of shortening displacement was male patients,and the variable that increased the risk of Z-axis rotation was the left limb.Shortening displacement was significantly positively correlated with lateral displacement(r=0.715,P<0.001);shortening displacement was significantly positively correlated with X-axis rotation displacement and Y-axis rotation displacement(r=0.265,P=0.028;r=0.303,P=0.011);lateral displacement was significantly positively correlated with Y-axis rotation and Z-axis rotation(r=0.258,P=0.032;r=0.250,P=0.038);X-axis rotation was significantly positively correlated with Y-axis rotation(r=0.382,P=0.001),and Z-axis rotation was significantly positively correlated with Y-axis rotation(r=0.280,P=0.020).(2)Postoperatively:The number of scapula fractures and comminuted bone fragments were variables that increased the risk of postoperative shortening and lateral displacement:Preoperative X-,Y-,and Z-axis rotation displacements were risk variables that increased postoperative X-,Y-,and Z-axis rotation displacements,respectively.Postoperative lateral displacement was significantly positively correlated with postoperative shortening and displacement(r=0.584,P=0.000),and postoperative lateral displacement was also significantly positively correlated with postoperative Y axis rotation and Z axis rotation(r=0.360,P=0.002;r=0.250,P=0.038).Postoperative Y axis rotation was significantly positively correlated with postoperative Z axis rotation(r=0.248,P=0.040).(3)The results showed that the three-dimensional displacement of the clavicle end before and after surgery was affected by many factors,especially the number of comminuted bone fragments,scapula fractures,gender,and original rotation displacement.At the same time,there were complex correlations between various displacements,especially the correlation between shortening displacement and lateral displacement was the strongest.
2.Three-dimensional displacement and risk factors of midshaft clavicle fractures treated with titanium elastic intramedullary nailing
Junwei ZHANG ; Lingling CHEN ; Zhenyuan MA ; Weizhi NIE ; Chaohui LI ; Haitao WANG ; Laibao DUAN ; Jinyong HOU ; Hongzheng BI
Chinese Journal of Tissue Engineering Research 2026;30(2):269-277
BACKGROUND:Titanium elastic intramedullary nailing for the treatment of significantly displaced midshaft clavicle fractures has the characteristics of minimally invasive and elastic fixation.The displacement of the fracture is closely related to the later function.However,there are few studies on the three-dimensional displacement analysis of the fracture ends before surgery and after intramedullary fixation such as titanium elastic intramedullary nailing.OBJECTIVE:To explore the three-dimensional displacement of fracture ends after midshaft clavicle fracture and fixation with titanium elastic intramedullary nails,and to analyze the risk factors.METHODS:A total of 91 patients with midshaft clavicle fracture(fracture end shortening ≥15 mm)admitted to Wendeng Orthopedic Hospital of Shandong Province from April 2019 to April 2024 were selected,including 57 males and 34 females,aged(51.73±10.21)years old.All patients received closed reduction and internal fixation with titanium elastic intramedullary nail.CT scans of the affected clavicle were performed before and on the first day after surgery.The CT data were imported into Mimics software for modeling.The length of the clavicle,lateral displacement of the fracture end,and rotation of the distal end of the fracture along the X,Y,and Z axes were measured and recorded before and after surgery.Pearson correlation coefficient was used for correlation analysis of various parameters,and generalized linear regression was used to evaluate risk factors.RESULTS AND CONCLUSION:(1)Preoperatively,the variable that increased the risk of lateral displacement was the number of comminuted bone fragments,the variable that increased the risk of shortening displacement was male patients,and the variable that increased the risk of Z-axis rotation was the left limb.Shortening displacement was significantly positively correlated with lateral displacement(r=0.715,P<0.001);shortening displacement was significantly positively correlated with X-axis rotation displacement and Y-axis rotation displacement(r=0.265,P=0.028;r=0.303,P=0.011);lateral displacement was significantly positively correlated with Y-axis rotation and Z-axis rotation(r=0.258,P=0.032;r=0.250,P=0.038);X-axis rotation was significantly positively correlated with Y-axis rotation(r=0.382,P=0.001),and Z-axis rotation was significantly positively correlated with Y-axis rotation(r=0.280,P=0.020).(2)Postoperatively:The number of scapula fractures and comminuted bone fragments were variables that increased the risk of postoperative shortening and lateral displacement:Preoperative X-,Y-,and Z-axis rotation displacements were risk variables that increased postoperative X-,Y-,and Z-axis rotation displacements,respectively.Postoperative lateral displacement was significantly positively correlated with postoperative shortening and displacement(r=0.584,P=0.000),and postoperative lateral displacement was also significantly positively correlated with postoperative Y axis rotation and Z axis rotation(r=0.360,P=0.002;r=0.250,P=0.038).Postoperative Y axis rotation was significantly positively correlated with postoperative Z axis rotation(r=0.248,P=0.040).(3)The results showed that the three-dimensional displacement of the clavicle end before and after surgery was affected by many factors,especially the number of comminuted bone fragments,scapula fractures,gender,and original rotation displacement.At the same time,there were complex correlations between various displacements,especially the correlation between shortening displacement and lateral displacement was the strongest.
3.Design, synthesis, and in vitro anti-tumor activity of silybin derivatives
Yan LI ; Lei GAO ; Chaohui ZHANG ; Yanqiu MENG
Journal of China Pharmaceutical University 2025;56(3):305-311
This study used the natural flavonoid compound silybin as the parent compound and synthesized 16 silybin derivatives through oxidative dehydrogenation, alkylation, selective demethylation, and acylation. The structures of these derivatives were confirmed by 1H NMR, 13C NMR, and MS. All derivatives were found to be new compounds never reported in previous literature. Using gastric cancer cell line SGC-7901 and human glioblastoma cell line LN-229, the in vitro anti-tumor activity of the novel silybin derivative was determined through MTT assay with lapatinib as the positive control. The experimental results indicate that the synthesized novel silybin derivatives have a certain degree of anti-proliferative effect on two types of cancer cells, with compounds I2 and I14 showing strong anti-proliferative activity against LN-229 and SGC-7901 cells.
4.Investigation of entrance surface doses from extra-oral dental X-ray devices in Nanping City, China
Jiahua TAN ; Haiyan CHU ; Yuanhao ZHANG ; Chaohui LI
Chinese Journal of Radiological Health 2025;34(5):751-757
Objective To investigate the entrance surface dose (ESD) of dental radiological diagnosis and treatment devices in Nanping City of Fujian Province, China, assess the radiation risk differences between cone beam computed tomography (CBCT) and conventional oral panoramic tomography (COPT), and compare the output dose levels between domestic and imported devices and between public and private medical institutions, and to provide evidence for optimizing radiation protection strategies. Methods A cross-sectional survey was conducted on 38 extraoral dental X-ray devices from 38 dental institutions in Nanping City. Measurements of ESD for different exposure types (adult/child COPT and CBCT) were performed using an adult head-and-neck phantom and thermoluminescent dosimeters. A statistical analysis (Mann-Whitney U test) was applied to compare dose differences by device type and institution type. Results The median tooth-region doses for adult/child CBCT (8.97/6.80 mGy) were significantly higher than those of adult/child COPT (0.33/0.24 mGy), with the 75th percentile values (12.44/8.18 mGy) exceeding the national guideline level of 7 mGy. Domestically manufactured devices demonstrated higher median doses and greater dose variability compared to imported devices (P < 0.05). Private institutions exhibited inferior dose control stability relative to public institutions (P < 0.001). CBCT delivered substantially higher radiation exposure to the thyroid and ocular lenses, with child lens dose reaching 5.9 times that of COPT. Conclusion Dental CBCT radiation doses in Nanping City exceed the guideline level. Enhanced protection for sensitive organs, restriction of unnecessary pediatric examinations, technological upgrades for domestic devices, and strengthened regulation and operational standardization of private institutions are critical to achieve precise radiation protection and improved medical quality.
5.Protective Effect of Ginkgo Biloba Extract on Damage of PV Interneurons in Schizophrenic Mice
Hongli ZHANG ; Kun WANG ; Chaohui DUAN ; Mingcai JI ; Sihan ZENG ; Xiaoyuan LYU
World Science and Technology-Modernization of Traditional Chinese Medicine 2025;27(5):1368-1375
Objective To investigate the mechanism of action of Ginkgo biloba extract on schizophrenia based on oxidative stress-mediated damage to PV interneurons.Methods 54 SPF grade male mice were selected as experimental subjects,divided into blank group,model group,ginkgo biloba extract 50 mg·kg-1,100 mg·kg-1,150 mg·kg-1 group,and risperidone group.Schizophrenic mouse models were established by intraperitoneal injection of MK-801 0.3 mg·kg-1,and behavioral(open field experiment,Y-maze,forced swimming)tests were conducted.Blood samples and brain tissue were collected 24 h after the last dose,Immunofluorescence staining was used to detect changes in PV neurons in the mouse brain;Detect the content of MAD,GSH Px,and SOD in serum using a reagent kit;ELISA method was used to detect the levels of iron and lipid peroxidation in mouse brain tissue;Western blot was used to detect the protein level of GPX4 in the mouse brain.Results Compared with the model group,the Ginkgo biloba leaf extract 150 mg·kg-1 group and the risperidone group significantly reduced the spontaneous alternation rate of the Y maze and significantly shortened the immobility time of forced swimming(P<0.05);PV neuron staining with varying degrees of enhanced fluorescence intensity;The MDA content in the serum of mice was significantly reduced(P<0.01),while the contents of SOD and GSH px were significantly increased(P<0.05);The iron content in the mouse brain was significantly reduced(P<0.05),the ROS content was significantly reduced(P<0.05),and the GPX4 content in the mouse brain was significantly increased(P<0.05).Conclusion Ginkgo biloba extract has a significant improvement effect on negative symptoms and cognitive impairment in MK-801 induced schizophrenia mouse models,and can also improve PV neuron damage in the prefrontal cortex of schizophrenia mice.Its mechanism may be related to the inhibition of iron death mediated oxidative stress by Ginkgo biloba extract.
6.Construction and evaluation of a diagnostic model for female stress urinary incontinence based on the mor-phology and elasticity of the levator ani muscle by transperineal three-dimensional ultrasound combined with shear wave elastography
Erfang GUO ; Lei FENG ; Chaohui SHI ; Ning LI ; Weiqun LIN ; Shuhua ZHANG
The Journal of Practical Medicine 2025;41(8):1224-1231
Objective To investigate the relationship between the morphology and elasticity of the levator ani muscle(LAM)and stress urinary incontinence(SUI),and to develop a multimodal diagnostic model for SUI based on LAM morphology and elasticity parameters,while evaluating the diagnostic performance of this model.Methods From September 2020 to September 2022,147 female patients with SUI from the Affiliated Hospital of North China University of Science and Technology were enrolled as the SUI group(case group),while 144 women without SUI during the same period were selected as the non-SUI group(control group).Transperineal ultrasonography was conducted to measure the anteroposterior diameter(LH-A1)and transverse diameter(LH-D1)of the levator hiatus at rest,the resting area of the levator hiatus(LA1),as well as the anteroposterior diameter(LH-A2),transverse diameter(LH-D2),and area(LA2)of the levator hiatus during the maximum Valsalva maneuver.Addi-tionally,ultrasonography was used to observe LAM injury(LA-MI)during pelvic muscle contraction.Shear wave elastography(SWE)was also performed transperineally to record the elastic modulus values of the puborectalis muscle at rest(E1)and during pelvic muscle contraction(E3).The differences in ultrasound parameters between the two groups were compared,and a logistic regression model was constructed for multivariate analysis to establish a diagnostic model for SUI.The goodness of fit of the logistic regression model was assessed using the Hosmer-Lemeshow test.The diagnostic performance of individual indicators and the diagnostic model for SUI was evaluated using the receiver operating characteristic(ROC)curve.Finally,the clinical utility of the model was assessed using decision curve analysis.Results There were statistically significant differences in age,BMI,LH-A1,LH-D1,LA1,LH-A2,LH-D2,LA2,LA-MI,E1,and E3 between the two groups(P<0.05).Multivariate logistic regression analysis revealed that age,BMI,LH-A1,LA2,LA-MI,E1,and E3 were significantly associated with SUI(P<0.05).Based on these findings,a diagnostic model for SUI was established:PRESUI=0.261×age+0.904×BMI-4.300×LH-A1+1.166×LA2-2.815×LA-MI+0.587×E1-0.631×E3-1.258.The model demon-strated excellent goodness-of-fit(P=0.983).The ROC curve analysis indicated that age,BMI,LH-A1,LA2,LA-MI,E1,and E3 all exhibited diagnostic efficacy for SUI(AUC>0.500,P<0.05).Notably,the AUC of the constructed diagnostic model for SUI was 0.996(95%CI:0.992~1.000),suggesting that the diagnostic accuracy of the model surpassed that of individual indicators.When the cut-off value of the diagnostic model was set at 0.437,the sensitivity reached 98.0%,and the specificity was 97.2%.Furthermore,the decision curve analysis demon-strated that the diagnostic model provided substantial net clinical benefit within the threshold probability range of 0.1 to 1.0.Conclusions The morphology and elasticity of the LAM are significantly altered in women with SUI.The SWE technique demonstrates potential application value for quantitatively assessing the elasticity of the LAM.Furthermore,the diagnostic model constructed based on age,BMI,LH-A1,LA2,LA-MI,E1,and E3 exhibits high clinical application value.
7.Construction and evaluation of a diagnostic model for female stress urinary incontinence based on the mor-phology and elasticity of the levator ani muscle by transperineal three-dimensional ultrasound combined with shear wave elastography
Erfang GUO ; Lei FENG ; Chaohui SHI ; Ning LI ; Weiqun LIN ; Shuhua ZHANG
The Journal of Practical Medicine 2025;41(8):1224-1231
Objective To investigate the relationship between the morphology and elasticity of the levator ani muscle(LAM)and stress urinary incontinence(SUI),and to develop a multimodal diagnostic model for SUI based on LAM morphology and elasticity parameters,while evaluating the diagnostic performance of this model.Methods From September 2020 to September 2022,147 female patients with SUI from the Affiliated Hospital of North China University of Science and Technology were enrolled as the SUI group(case group),while 144 women without SUI during the same period were selected as the non-SUI group(control group).Transperineal ultrasonography was conducted to measure the anteroposterior diameter(LH-A1)and transverse diameter(LH-D1)of the levator hiatus at rest,the resting area of the levator hiatus(LA1),as well as the anteroposterior diameter(LH-A2),transverse diameter(LH-D2),and area(LA2)of the levator hiatus during the maximum Valsalva maneuver.Addi-tionally,ultrasonography was used to observe LAM injury(LA-MI)during pelvic muscle contraction.Shear wave elastography(SWE)was also performed transperineally to record the elastic modulus values of the puborectalis muscle at rest(E1)and during pelvic muscle contraction(E3).The differences in ultrasound parameters between the two groups were compared,and a logistic regression model was constructed for multivariate analysis to establish a diagnostic model for SUI.The goodness of fit of the logistic regression model was assessed using the Hosmer-Lemeshow test.The diagnostic performance of individual indicators and the diagnostic model for SUI was evaluated using the receiver operating characteristic(ROC)curve.Finally,the clinical utility of the model was assessed using decision curve analysis.Results There were statistically significant differences in age,BMI,LH-A1,LH-D1,LA1,LH-A2,LH-D2,LA2,LA-MI,E1,and E3 between the two groups(P<0.05).Multivariate logistic regression analysis revealed that age,BMI,LH-A1,LA2,LA-MI,E1,and E3 were significantly associated with SUI(P<0.05).Based on these findings,a diagnostic model for SUI was established:PRESUI=0.261×age+0.904×BMI-4.300×LH-A1+1.166×LA2-2.815×LA-MI+0.587×E1-0.631×E3-1.258.The model demon-strated excellent goodness-of-fit(P=0.983).The ROC curve analysis indicated that age,BMI,LH-A1,LA2,LA-MI,E1,and E3 all exhibited diagnostic efficacy for SUI(AUC>0.500,P<0.05).Notably,the AUC of the constructed diagnostic model for SUI was 0.996(95%CI:0.992~1.000),suggesting that the diagnostic accuracy of the model surpassed that of individual indicators.When the cut-off value of the diagnostic model was set at 0.437,the sensitivity reached 98.0%,and the specificity was 97.2%.Furthermore,the decision curve analysis demon-strated that the diagnostic model provided substantial net clinical benefit within the threshold probability range of 0.1 to 1.0.Conclusions The morphology and elasticity of the LAM are significantly altered in women with SUI.The SWE technique demonstrates potential application value for quantitatively assessing the elasticity of the LAM.Furthermore,the diagnostic model constructed based on age,BMI,LH-A1,LA2,LA-MI,E1,and E3 exhibits high clinical application value.
8.Long-term oncological safety of robotic total gastrectomy for locally advanced proximal gastric cancer: a 5-year noninferiority comparison based on the FUGES-014 study
Qing ZHONG ; Zhiquan ZHANG ; Yongqi YAN ; Yifan LI ; Qichen HE ; Chaohui ZHENG ; Qiyue CHEN ; Changming HUANG
Chinese Journal of Gastrointestinal Surgery 2025;28(8):886-894
Objective:To report the 5-year survival outcomes and recurrence patterns of robotic total gastrectomy (RTG) for locally advanced proximal gastric cancer in order to provide more valuable long-term follow-up results for clinical practice.Methods:This was a prospective, single-arm, open-label clinical trial (FUGES-014; Clinical-Trials.gov, NCT03524287). Patients with locally advanced proximal gastric cancer who underwent RTG at Fujian Medical University Union Hospital from March 5, 2018, to February 10, 2020, were included in the analysis. To evaluate the long-term efficacy of RTG in the most objective manner possible, we performed a propensity score-matched (1∶2) comparative analysis with historical control patients who had undergone laparoscopic total gastrectomy (LTG) from the FUGES-002 study (ClinicalTrials.gov, NCT02333721) in which the 5-year disease-free survival (DFS), 5-year overall survival (OS), and recurrence patterns were compared between the two groups.Results:Prior to matching, there were 48 cases in the RTG group and 263 cases in the LTG group; patients in the LTG group had more advanced cT and pT stages ( P=0.044 and 0.006, respectively) compared to the RTG group. After matching, there were 48 cases in the RTG group and 96 cases in the LTG group; however, no statistically significant differences were observed in the baseline clinical characteristics between the two groups (all P>0.05). Both groups had a median follow-up of 72 months. The 5-year DFS rates were 75.0% (95%CI: 63.7%- 88.3%) in the RTG group and 61.4% (95%CI: 52.5%-72.0%) in the LTG group ( P=0.116). Similarly, the 5-year OS rates were 79.2% (95%CI: 68.5%-91.5%) and 64.6% (95%CI: 55.7%-74.9%) in the RTG and LTG groups, respectively ( P=0.100). Within 5 years after surgery, tumor recurrence occurred in 10 patients (20.8%) in the RTG group and 33 patients (34.4%) in the LTG group ( P=0.124), and peritoneal recurrence was the predominant pattern in both groups (8.3%[4/48] vs. 10.4%[10/96]; risk difference: -0.02, P=0.554). Gastric cancer-related death was the predominant cause of death in both groups (16.7% [8/48] vs. 31.2% [30/96]; risk difference: -0.15, P=0.064). Among patients stratified by different pathological stages, no statistically significant differences were found in DFS, OS, or recurrence rates between the RTG and LTG groups (all P>0.05). Conclusions:We find the long-term oncological outcomes of RTG for locally advanced proximal gastric cancer to be noninferior to those of LTG. RTG should therefore be considered as a valid option for standardized minimally invasive surgery for locally advanced proximal gastric cancer.
9.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.
10.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.

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