1.Construction and biological characteristics study of RuvB gene deletion strain of Glaesserella parasuis
Jiancheng ZHAO ; Ling HUAGN ; Lihua XU ; Xiufang YUAN ; Bin YU ; Fei SU ; Shiyi YE ; Hongchao SUN ; Hui ZHANG ; Junxing LI
Chinese Journal of Veterinary Science 2025;45(11):2358-2364
Glaesserella parasuis is the causative agent of Gl?sser's disease in pigs.However,the pathogenic mechanisms underlying its virulence is not yet fully understood.The RuvB protein,a member of the AAA+superfamily,is implicated in various cellular processes,yet its specific role in the virulence of Glaesserella parasuis has not been fully characterized.In this study,we con-structed a RuvB gene deletion mutant,designated ΔRuvB,using the serotype 13 Glaesserella pa-rasuis strain ZJ1208 and a suicide plasmid-mediated natural transformation approach.To elucidate the functional role of the RuvB gene,we comprehensively evaluated the biological characteristics of the ΔRuvB strain through a series of assays,including growth kinetics,colony morphology,bac-terial staining,transmission electron microscopy(TEM),osmotic stress tolerance,high-tempera-ture tolerance,heat shock resistance,UV resistance,capsular polysaccharide quantification,serum bactericidal assays,and murine virulence experiments.Our findings revealed that the growth rate of ΔRuvB showed no significant difference compared to the parental strain.TEM revealed a notable increase in bacterial cell length;however,the number of outer membrane vesicles(OMVs)on the surface of ΔRuvB did not significantly increase.Notably,the ΔRuvB strain displayed a significant reduction in capsular polysaccharide production and serum resistance,as well as diminished toler-ance to UV radiation and high temperatures.Significant alterations were observed in its resistance to osmotic stress or oxidative stress.In the mouse toxicity challenge experiment,in com-parison with the parental strain ZJ1208,the mortality rate dropped by 20 percentage points,suggesting that the virulence of ΔRuvB has been weakened to some extent.Collectively,these results underscore the critical role of the RuvB gene in enhancing the environmental adaptability of Glaesserella parasuis.
2.Serotyping,virulence factors and drug resistance analysis of Pasteurella multocida of porcine origin of Zhejiang Province
Yizhen XU ; Xiufang YUAN ; Lihua XU ; Bin YU ; Fei SU ; Shiyi YE ; Hongchao SUN ; Yijie CHEN ; Qiaodan CHEN ; Hui ZHANG ; Junxing LI
Chinese Journal of Veterinary Science 2025;45(7):1378-1387
The aim of this paper was to investigate the serotypes,virulence factors and drug resist-ance of clinical isolates of Pasteurella rnultocida of porcine origin in recent years.Morphological screening and polymerase chain reaction(PCR)were used to isolate and identify 119 isolates from nasal swabs and lung tissue samples sent from swine farms in Zhejiang Province from 2010 to 2024.The isolates of Pasteurella multocida were subjected to capsular polysaccharide serotyping,lipopolysaccharide serotyping,virulence factor detection and drug resistance analysis by PCR and Kirby-Bauer disc agar diffusion method(K-B).The results showed that there were 64 strains(53.7%)of A-type,54 strains(45.3%)of D-type and 1 strain(0.9%)of F-type among the capsu-lar polysaccharide serotypes,and 10 strains(8.4%)of L1-type,20 strains(16.8%)of L3-type,86 strains(72.2%)of L6-type,and 3 strains(2.6%)of undetermined type among the lipopolysaccha-ride serotypes.The amplification results of 10 virulence genes showed that the detection rate of virulence genes hgbA,higbB and fimA was over 86.0%,the detection rate of toxA was 8.4%,while the virulence gene tbpA was not detected.There were also differences in the distribution vir-ulence genes in different capsular polysaccharide serotypes.Virulence factor pfhA was detected in type A and F but not in type D.The detection rate of adhesin gene tadD in serotype A(92.2%)was significantly higher than that of type D(9.3%),and,on the contrary,the detection rate of ad-hesin gene hsf-l in serotype D(90.7%)was significantly higher than that of type A(20.3%).Drug resistance analysis revealed that Pasteurella multocida showed high susceptibility to antimi-crobial drugs such as amoxicillin,ampicillin,cephalosporins,doxycycline,fosfenicol and ciprofloxa-cin,and showed strong resistance to antimicrobial drugs such as lincomycin,cotrimoxazole,genta-micin and amikacin,and there were 54 multi-drug resistant strains(78.3%).In summary,capsular polysaccharide serotypes were dominated by type A and D,lipopolysaccharide serotypes were dom-inated by L6,the distribution of some virulence genes varied greatly among different serotypes,and the proportion of multi-resistant strains was high,which provide reference for the prevention and control of this disease.
3.Construction and biological characteristics study of RuvB gene deletion strain of Glaesserella parasuis
Jiancheng ZHAO ; Ling HUAGN ; Lihua XU ; Xiufang YUAN ; Bin YU ; Fei SU ; Shiyi YE ; Hongchao SUN ; Hui ZHANG ; Junxing LI
Chinese Journal of Veterinary Science 2025;45(11):2358-2364
Glaesserella parasuis is the causative agent of Gl?sser's disease in pigs.However,the pathogenic mechanisms underlying its virulence is not yet fully understood.The RuvB protein,a member of the AAA+superfamily,is implicated in various cellular processes,yet its specific role in the virulence of Glaesserella parasuis has not been fully characterized.In this study,we con-structed a RuvB gene deletion mutant,designated ΔRuvB,using the serotype 13 Glaesserella pa-rasuis strain ZJ1208 and a suicide plasmid-mediated natural transformation approach.To elucidate the functional role of the RuvB gene,we comprehensively evaluated the biological characteristics of the ΔRuvB strain through a series of assays,including growth kinetics,colony morphology,bac-terial staining,transmission electron microscopy(TEM),osmotic stress tolerance,high-tempera-ture tolerance,heat shock resistance,UV resistance,capsular polysaccharide quantification,serum bactericidal assays,and murine virulence experiments.Our findings revealed that the growth rate of ΔRuvB showed no significant difference compared to the parental strain.TEM revealed a notable increase in bacterial cell length;however,the number of outer membrane vesicles(OMVs)on the surface of ΔRuvB did not significantly increase.Notably,the ΔRuvB strain displayed a significant reduction in capsular polysaccharide production and serum resistance,as well as diminished toler-ance to UV radiation and high temperatures.Significant alterations were observed in its resistance to osmotic stress or oxidative stress.In the mouse toxicity challenge experiment,in com-parison with the parental strain ZJ1208,the mortality rate dropped by 20 percentage points,suggesting that the virulence of ΔRuvB has been weakened to some extent.Collectively,these results underscore the critical role of the RuvB gene in enhancing the environmental adaptability of Glaesserella parasuis.
4.Serotyping,virulence factors and drug resistance analysis of Pasteurella multocida of porcine origin of Zhejiang Province
Yizhen XU ; Xiufang YUAN ; Lihua XU ; Bin YU ; Fei SU ; Shiyi YE ; Hongchao SUN ; Yijie CHEN ; Qiaodan CHEN ; Hui ZHANG ; Junxing LI
Chinese Journal of Veterinary Science 2025;45(7):1378-1387
The aim of this paper was to investigate the serotypes,virulence factors and drug resist-ance of clinical isolates of Pasteurella rnultocida of porcine origin in recent years.Morphological screening and polymerase chain reaction(PCR)were used to isolate and identify 119 isolates from nasal swabs and lung tissue samples sent from swine farms in Zhejiang Province from 2010 to 2024.The isolates of Pasteurella multocida were subjected to capsular polysaccharide serotyping,lipopolysaccharide serotyping,virulence factor detection and drug resistance analysis by PCR and Kirby-Bauer disc agar diffusion method(K-B).The results showed that there were 64 strains(53.7%)of A-type,54 strains(45.3%)of D-type and 1 strain(0.9%)of F-type among the capsu-lar polysaccharide serotypes,and 10 strains(8.4%)of L1-type,20 strains(16.8%)of L3-type,86 strains(72.2%)of L6-type,and 3 strains(2.6%)of undetermined type among the lipopolysaccha-ride serotypes.The amplification results of 10 virulence genes showed that the detection rate of virulence genes hgbA,higbB and fimA was over 86.0%,the detection rate of toxA was 8.4%,while the virulence gene tbpA was not detected.There were also differences in the distribution vir-ulence genes in different capsular polysaccharide serotypes.Virulence factor pfhA was detected in type A and F but not in type D.The detection rate of adhesin gene tadD in serotype A(92.2%)was significantly higher than that of type D(9.3%),and,on the contrary,the detection rate of ad-hesin gene hsf-l in serotype D(90.7%)was significantly higher than that of type A(20.3%).Drug resistance analysis revealed that Pasteurella multocida showed high susceptibility to antimi-crobial drugs such as amoxicillin,ampicillin,cephalosporins,doxycycline,fosfenicol and ciprofloxa-cin,and showed strong resistance to antimicrobial drugs such as lincomycin,cotrimoxazole,genta-micin and amikacin,and there were 54 multi-drug resistant strains(78.3%).In summary,capsular polysaccharide serotypes were dominated by type A and D,lipopolysaccharide serotypes were dom-inated by L6,the distribution of some virulence genes varied greatly among different serotypes,and the proportion of multi-resistant strains was high,which provide reference for the prevention and control of this disease.
5.Development of a mobile personnel radiation protection equipment
Yuan HE ; Hongchao PANG ; Yang LIU ; Jianhua WU ; Junlin WANG ; Yanan HONG ; Xubiao CHEN ; Hualin WANG ; Xiaoyu SHI ; Miao PAN ; Qiang WANG
Chinese Journal of Radiological Health 2024;33(2):158-163
Objective To design a mobile personnel radiation protection equipment for operation in environments with high radiation such as spent fuel reprocessing plants, to achieve simultaneous protection against γ radiation, neutron radiation, and radioactive aerosol, to reduce the internal and external exposure dose of radioactive workers, and to meet the requirement of operation for two hours. Methods The core parts of the mobile personnel radiation protection equipment included a shielding chamber and a respiratory maintenance system. An automated chassis was used for the movement and lifting of the shielding chamber. MCNP software was used to simulate and calculate the protective effects of shielding chamber made of different materials and material thicknesses. Experimental verification of the shielding chamber design was conducted. Mathematical models were established to describe the variations in the content of various gases in the chamber with personnel operation time. A respiratory maintenance system, a harmful gas absorption device, and an automated mobile chassis were designed. Results The shielding chamber made of polyethylene with a thickness of 80 mm achieved an 80% neutron shielding rate. The respiratory maintenance system could support workers for 2 hours of operation inside the equipment. The mobile chassis allowed operation of the equipment with one person. Conclusion This mobile personnel radiation protection equipment can solve the problem in simultaneous protection against γ radiation, neutron radiation, and radioactive aerosol. The equipment can provide radiation protection for radioactive workers, reduce exposure dose, and reduce personnel burden. This system provides technical means for the operation and maintenance of equipment in high-radiation sites such as spent fuel reprocessing plants.
6.Development of a simplified phantom for calibration of whole-body counter
Jianhua WU ; Hongchao PANG ; Zhiping LUO ; Yang LIU ; Xinglong LI ; Guojun YUAN
Chinese Journal of Radiological Health 2023;32(3):259-263
Objective To develop a simplified phantom for the calibration of whole-body counters. Methods A simplified phantom design method for the calibration of whole-body counters was established based on the process and method of calibrating whole-body counters. By using the established method and Monte Carlo method, a simplified phantom including the total body, thyroid, lungs, and gastrointestinal tract was designed to calibrate the ORTEC-Stand FAST II whole-body counter. The simplified phantom was compared with the BOMAB phantom through experimental measurements. Results Within the range of 50 keV to 2 MeV, for rays of the same energy in the same organ of the simplified phantom and BOMAB phantom, the simulated data of detection efficiency by whole-body counting showed an error within 5%, and the experimental measurements showed an error within 10%. Conclusion We developed a simplified phantom for the calibration of the whole-body counter, demonstrating the feasibility of using the simplified phantom instead of a physical body phantom for whole-body counter calibration, which can greatly facilitate whole-body counter calibration for internal radiation monitoring.
7.Research progress on the biotransformation of flavonoid glycosides in Epimedii Folium
Yuhang ZHANG ; Wang CHEN ; Zili FENG ; Hongchao YUAN ; Xiaolin GAO ; Cuiping WANG
China Pharmacy 2022;33(12):1525-1529
Flavonoid glycosides are the main active constituents of Epimedii Folium and its related plants. They can be divided into polyglycosides and low glycosides according to the number of glycosyl group. The polyglycosides of Epimedii Folium can be transformed into low glycosides after biotransformation ;pharmacological activities of low glycosides in anti-tumor ,tonifying kidney yang and anti-osteoporosis are stronger than those of polyglycosides. In this paper , the research progress about biotransformation technology of flavonoid glycosides of Epimedii Folium was reviewed. It was found that the main biotransformation pathway of flavonoid glycosides of Epimedii Folium was to obtain low glycosides by removing glycosyl group ; related methods were mainly enzymatic hydrolysis and microbial transformation ,and also included plant cell transformation ,acid hydrolysis method and synthesis method.
8.Association between human papilloma virus 16 E6 and E7-specific T cell immune response and clinical prognosis of patients with cervical squamous cell carcinoma
Hongchao CAI ; Yuanliang YUAN ; Alifu MAYINUER ; Ruozheng WANG
Chinese Journal of Radiation Oncology 2021;30(4):357-362
Objective:To investigate the relationship between human papilloma virus (HPV)16 E6/7-specific T cell immune response in the periphral blood and clinical features and prognosis of patients with cervical squamous cell carcinoma (CSCC).Methods:Seventy-two patients pathologically diagnosed with CSCC admitted to Affiliated Tumor Hospital of Xinjiang Medical University from June 2013 to October 2015, and 75 healthy controls were enrolled in this study. The special responses of peripheral blood T cells to E6 and E7 overlapping peptides before treatment were detected by enzyme-linked immunosorbent assay (ELISA). The differences of frequency and intensity expression of specific immune responses between two groups were analyzed by chi-square χ2 test and nonparametric test. The correlation between antigen-specific immune response and T cell subsets was analyzed by Spearman test. Log-rank test and Cox’s regression model were employed for univariate and multivariate prognostic analyses. Results:The frequencies of HPV16 E6-ad E7-specific T cell responses in CSCC patients were significantly higher than those in healthy controls (51.39% vs. 29.33%, P=0.006 and 45.83% vs.25.33%, P=0.009), and the mean intensities were also considerably higher than those in healthy controls (20.00 SFC/10 6vs.10.76 SFC/10 6, P<0.001 and 16.17 SFC/10 6vs.10.72 SFC/10 6, P=0.017). The intensity of HPV16 E6-specific T cell immune response was positively correlated with the CD 4+/CD 8+ ratio in the peripheral blood of CSCC patients ( r=0.279, P=0.018). And a strong correlation was noted between E7-specific T cell immune response intensity and increasing proportion of NK+ cells ( r=0.274, P=0.020). Univariate and multivariate analyses showed that therapeutic mode (radiotherapy vs. concurrent chemoradiotherapy, HR=2.918, 95% CI 1.454-5.854, P=0.003) and E6-specific T cell response (response group vs. no response group, HR=0.491, 95% CI 0.243-0.99, P=0.047) were the independent prognostic factors influencing the clinical prognosis. The 5-year overall survival in patients with HPV16 E6-specific T cell responses was significantly higher than that in the no response group (64% vs.41%, P=0.041). Conclusions:The intensity of HPV16 E6-specific T cell immune response is positively correlated with the CD 4+/CD 8+ ratio. No HPV16 E6-specific T cell response and radiotherapy alone are more likely to cause poor prognosis of CSCC patients.
9.Effects of morphological characteristics of femoral intertrochanteric fractures on internal fixation
Shuo WANG ; 300191 中国人民解放军464医院骨科 ; anxiong Ji MA ; Xinlong MA ; Yuan XUE ; Hongchao HUANG ; Hongwei GU ; Shangqi JIAO
Chinese Journal of Orthopaedics 2017;37(18):1163-1170
Objective To explore the effects of internal fixation in surgical treatment of intertrochanteric fractures by proximal femoral locking compression plate (PFLCP) and proximal femoral nail anti-rotation (PFNA) on fracture morphological fea-tures. Methods 43 patients with femoral intertrochanteric fractures were treated with PFLCP or PFNA from January 2013 to De-cember 2015. There were 23 patients treated with PFLCP and 20 cases with PFNA. There were 29 males and 14 females, the aver-age age was 62.6 years old (ranged from 46-85 years). Observed the fracture type (AO/OTA, Evans, the lateral trochanteric wall in-jury), the direction of key fracture line (lateral superior to medial inferior/medial superior to lateral inferior), preoperative days, pre-operative bone mineral density value, operation time, the volume of intraoperative blood loss, the supportive contact rate at the me-dial cortex of proximal femur after surgery, the surgical intervention rate for medial cortical fracture, postoperative partial load bearing time, full weight bearing time, complications of internal fixation and Harris scores. And analyzed the relationship between the direction of key fracture line and the internal fixation failure. SPSS13.0 software was used for statistical analysis. Results No statistical significance was found in age distribution, fracture types, preoperative days, preoperative bone mineral density value, the internal fixation failure rate (loss of fracture reduction, screw cutting or plate fracture). The operative time of PFLCP group was obviously longer than that of PFNA group and the difference was statistically significant (t=2.216, P=0.032). The intraoperative blood loss was more in PFLCP group than PFNA group and significant difference was found between the two groups(t=4.142, P=0.000).Though the initial postoperative partial load bearing time is earlier in the PFNA group than PFLCP group, but no statistical difference bewtween the two groups (t=0.833, P=0.902). Full weight bearing time had no statistical difference between the two groups. Harris scores in the PFNA group was slightly lower than in the PFLCP group, but there was no statistical difference ( t=0.833, P=0.902). The postoperative medial cortical support rate was higher in the PFLCP group than PFNA group, but no statisti-cal difference between them. The surgical intervention rate for medial cortical fracture was higher in the PFLCP group and differ-ence was statistically significant (χ2=4.768, P=0.029). No correlation had found between invalid support at medial cortex and the direction of key fracture line in all internal fixation failure cases. Distribution of the direction of key fracture line in the two groups had no statistical difference, while the relative risk ratio in terms of internal fixation failure between 2 types of the direction of key fracture line (OR=lateral superior to medial inferior/ medial superior to lateral inferior) had statistical difference (χ2=6.081, P=0.014;OR=9.600, P=0.037). Conclusion The direction of key fracture line can foresee the underlying displacement direction of intertrochanteric fractures at postsurgical load bearing, combining with restore the supportive contact at the medial cortex of the proximal femur and timing of load bearing may be determined to avoid internal fixation failure.
10.Evaluation of the early clinical effect of medical calcium sulfate in traumatic fracture treatment
Shuo WANG ; Jianxiong MA ; Xinlong MA ; Yuren DU ; Yuan XUE ; Hongchao HUANG ; Lei ZHANG
The Journal of Practical Medicine 2017;33(14):2299-2302
Objective To investigate the X-ray gray scale changes of calcium sulfate and evaluate its clini-cal effect in traumatic fracture treatment. Methods 23 traumatic fracture cases were treated from September 2014 to January 2016 in our hospital. The degradation rate of calcium sulfate was evaluated by X ray assay. Results Af-ter surgery,about 69%remnants at 1 week,53%remnants at 2 weeks,26%remnants at 4 weeks,7%remnants at 6 weeks were observed,while no remnants were found at 8 weeks after surgery. The initial time window of callus appearance was 3 to 9 weeks and the mean time was(6.5 ± 1.6)weeks. The fracture union time was 8 to 24 weeks and the mean time was(15.0 ± 5.2)weeks. One patient with distal humeral comminuted fracture had non-infec-tious delayed healing wound.One case of hemolytic staphylococcus in incision was cultured. Conclusion Calcium sulfate degrades rapidly,cautions should be taken for the application in the superficial bone.

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